Genomics, Proteomics & Bioinformatics•2024•DOI: 10.1093/gpb/art_1123
Gametogenesis plays an important role in the reproduction and evolution of species. The transcriptomic and epigenetic alterations in this process can influence the reproductive capacity, fertilization, and embryonic development. The rapidly increasing single-cell studies have provided valuable multi-omics resources. However, data from different layers and sequencing platforms have not been uniformed and integrated, which greatly limits their use for exploring the molecular mechanisms that underlie oogenesis and spermatogenesis. Here, we develop GametesOmics, a comprehensive database that integrates the data of gene expression, DNA methylation, and chromatin accessibility during oogenesis and spermatogenesis in humans and mice. GametesOmics provides a user-friendly website and various tools, including Search and Advanced Search for querying the expression and epigenetic modification(s) of each gene; Tools with Differentially expressed gene (DEG) analysis for identifying DEGs, Correlation analysis for demonstrating the genetic and epigenetic changes, Visualization for displaying single-cell clusters and screening marker genes as well as master transcription factors (TFs), and MethylView for studying the genomic distribution of epigenetic modifications. GametesOmics also provides Genome Browser and Ortholog for tracking and comparing gene expression, DNA methylation, and chromatin accessibility between humans and mice. GametesOmics offers a comprehensive resource for biologists and clinicians to decipher the cell fate transition in germ cell development, and can be accessed at http://gametesomics.cn/.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04245-1
Background Chronic limb-threatening ischemia (CLTI) is the most severe form of peripheral arterial disease (PAD). Mesenchymal stem cell (MSC) transplantation holds promise as a treatment for CLTI; however, the harsh local environment poses challenges to its effectiveness. Apoptotic vesicles (ApoVs) are extracellular vesicles produced by cells undergoing apoptosis, and they can carry various biomolecules from their parent cells, including proteins, RNA, DNA, lipids, ions, and gas neurotransmitters. ApoVs play significant roles in anti-inflammatory responses, anti-tumor activities, and tissue regeneration through intercellular communication, and they have demonstrated potential as drug carriers. In this study, we investigated the potential of bone marrow stem cell (BMSC)-derived ApoVs for treating CLTI. Methods In vivo, we explored the therapeutic effect of ApoVs on a hindlimb ischemia model through Laser Doppler, matrigel plug assay, and histological analysis. In vitro, we analyzed the effects of ApoVs on the proliferation, migration, and angiogenesis of HUVECs and explored the uptake process of ApoVs. In addition, Proteomic analysis, western blotting, quantitative real-time PCR, shRNA, and siRNA were used to analyze ApoVs-induced HUVECs activation and downstream signaling pathways. Results BMSCs transplantation showed improvement in a hind limb ischemia model, and this effect still exists after apoptosis of BMSCs. Subsequently, ApoVs of BMSCs were isolated and found to improve mouse hind limb ischemia in vivo. In vitro, ApoVs can be ingested by HUVECs through dynamin-, clathrin-, and caveolin-mediated endocytosis and promote its proliferation, migration, and angiogenesis. Mechanistically, ApoVs transferred NAMPT to HUVECs, therefore activating the NAMPT/SIRT1/FOXO1 axis, influencing the transcriptional activity of FOXO1, and promoting angiogenesis. Conclusions Our results demonstrate that the transplanted BMSCs can ameliorate hindlimb ischemia by releasing ApoVs during apoptosis. The main mechanism of this effect is promoting the proliferation, migration, and angiogenesis of endothelial cells via the NAMPT/SIRT1/FOXO1 axis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04131-w
Background Alzheimer’s disease (AD) is a progressive neurodegenerative condition affecting around 50 million people worldwide. Bone marrow-derived mesenchymal stem cells (BMMSCs) have emerged as a promising source for cellular therapy due to their ability to differentiate into multiple cell types and their paracrine effects. However, the direct injection of BMMSCs can lead to potential unpredictable impairments, prompting a renewed interest in their paracrine effects for AD treatment. The specific mechanism and central role of cytokines in this process have not been fully elucidated. Methods Mouse BMMSCs were isolated, validated, and then transplanted intracerebrally into APP/PS1 female mice. The behavioral tests, including open-field test, novel object recognition test, and Morris water maze were performed, followed by β-amyloidosis plaque and neuron apoptosis analyses. Then the tissue RNA sequencing and mBMMSC cytokine analysis were performed. A cytokine antibody array for BMMSCs and the brain slice models were performed with AD model tissues were used to elucidate the molecular mechanisms. Finally, APP/PS1 mice were administrated with cytokine mixture for cognitive recovery. Results Our results demonstrated that BMMSCs significantly improved cognitive function, reduced beta-amyloid plaque deposition, and decreased apoptotic neurons through the activation of the AKT signaling pathway. Using a cytokine antibody array, we identified three highly expressed AKT pathway regulated neuroprotective factors in BMMSCs: IGF1, VEGF, and Periostin2. These cytokines were found to upregulate inhibitors of apoptosis family proteins (IAPs) and suppress Caspase-3 activity in brain slices induced with beta amyloidosis (Aβ), okadaic acid (OA), and lipopolysaccharide (LPS). When injection of this cytokine mixture to APP/PS1 mice also resulted in a mitigation of cognitive impairment. Conclusions These findings suggest that the secretory factors IGF1, VEGF, and Periostin2 derived from BMMSCs play a crucial role in neuroprotection by modulating the AKT/IAPs pathway to restore neuronal function. These cytokine sets could be a potential therapeutic strategy for AD and lay the groundwork for promising clinical applications.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03886-y
Background Understanding the lineage differentiation of human prostate not only is crucial for basic research on human developmental biology but also significantly contributes to the management of prostate-related disorders. Current knowledge mainly relies on studies on rodent models, lacking human-derived alternatives despite clinical samples may provide a snapshot at certain stage. Human embryonic stem cells can generate all the embryonic lineages including the prostate, and indeed a few studies demonstrate such possibility based on co-culture or co-transplantation with urogenital mesenchyme into mouse renal capsule. Methods To establish a stepwise protocol to obtain prostatic organoids in vitro from human embryonic stem cells, we apply chemicals and growth factors by mimicking the regulation network of transcription factors and signal transduction pathways, and construct cell lines carrying an inducible NKX3-1 expressing cassette, together with three-dimensional culture system. Unpaired t test was applied for statistical analyses. Results We first successfully generate the definitive endoderm, hindgut, and urogenital sinus cells. The embryonic stem cell-derived urogenital sinus cells express prostatic key transcription factors AR and FOXA1, but fail to express NKX3-1. Therefore, we construct NKX3-1-inducible cell line by homologous recombination, which is eventually able to yield AR, FOXA1, and NKX3-1 triple-positive urogenital prostatic lineage cells through stepwise differentiation. Finally, combined with 3D culture we successfully derive prostate-like organoids with certain structures and prostatic cell populations. Conclusions This study reveals the crucial role of NKX3-1 in prostatic differentiation and offers the inducible NKX3-1 cell line, as well as provides a stepwise differentiation protocol to generate human prostate-like organoids, which should facilitate the studies on prostate development and disease pathogenesis.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025207
O-GlcNAcylation, a prevalent reversible post-translational modification, intricately alters non-histone proteins, influencing the organization of gene transcriptional regulation within the accessible chromatin environment. This nucleoplasmic landscape, characterized by histone-free regions, fundamentally enables O-GlcNAc-mediated modulation through dynamic accessibility. However, unraveling the O-GlcNAc-open chromatin interplay that governs sophisticated transcriptional regulatory networks remains constrained by current techniques, which lack the resolution to probe this spatiotemporal crosstalk. Here, we report a general strategy to systematically and chemoselectively profile O-GlcNAc-associated chromatin accessibility on a genome-wide scale (COCA-seq). Through comprehensive validation across low- and high-throughput levels, we demonstrate COCA-seq’s dual fidelity in both O-GlcNAc chemoselectivity and open chromatin specificity. We employ it to delve into doxorubicin resistance for breast cancer, scrutinizing pivotal regulatory genes and transcription factors implicated in this complex biological event. By integrating bulk RNA-seq with COCA-seq, we offer a multiomics perspective, shedding light on related biological processes and pathways like drug efflux and stress homeostasis, thereby uncovering potential mechanisms by which O-GlcNAc-associated open chromatin orchestrates tumor drug resistance. COCA-seq emerges as a general and versatile tool across various biological contexts, poised to reveal the landscape of O-GlcNAc-associated open chromatin regions across the genome and decipher the significance of glycosylation behind it.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024183
Traumatic brain injury (TBI) is a recognized global public health problem. However, there are still limitations in the available therapeutic approaches and a lack of clinically effective drugs. Therefore, an in-depth exploration of the secondary pathological mechanism of TBI and the identification of new effective drugs are urgently needed. Cannabidiol (CBD), a component derived from the cannabis plant, has potential therapeutic effects on neurological diseases and has received increasing attention. However, few reports on CBD intervention in TBI patients exist. Here, we use the Feeney free-fall method to establish a rat TBI model. CBD significantly improves neurological deficit scores, neuronal damage and blood-brain barrier permeability in rats and significantly inhibits the expressions of the brain injury markers S-100β and NSE. Mechanistically, CBD attenuates TBI-induced astrocyte activation, reduces inflammation, and attenuates the expressions of inflammatory prostaglandin system indicators. The use of TG6-10-1 (EP2 inhibitor) and H-89 (PKA inhibitor) indicates that CBD attenuates TBI-induced neurological damage via the PGE2-EP2-cAMP-PKA signaling pathway. Overall, this research provides a novel drug candidate for the treatment of clinical brain trauma.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025114
Bladder cancer (BCa) is one of the most common malignant tumors of the urinary system, but its pathogenesis is still unclear. T1G3 BCa is particularly invasive and relapses readily after treatment, with progression to invasive cancer or distant metastasis. Therefore, identification of the molecular mechanism by which it invades and metastasizes to guide treatment and predict patient prognosis is needed. Cofilin1 plays an important role in regulating gene expression and the invasiveness of tumors. In this study, we show that Cofilin1 is highly expressed in BCa and lymph nodes with metastasis, which is positively related to the grade of BCa, and is significantly related to clinicopathological parameters and cancer-specific survival. Phenotypic analysis reveals that Cofilin1 knockout inhibits the proliferation and migration of BCa cells, whereas Cofilin1 overexpression promotes the opposite phenotype. Cofilin1 binds to cortactin, thereby reducing the expression of F-actin and promoting the formation of invadopodia in BCa cells. Further experiments reveal that TCF7L2 can bind to the promoter of Cofilin1 and transactivate it, promoting a malignant phenotype. TCF7L2 may also reverse the inhibitory effect of miR-206 on the binding of Cofilin1 and cortactin and promote the metastasis of BCa by inhibiting the transcription maturation of miR-206. This study confirms that Cofilin1 is an oncogene in T1G3 BCa, and the TCF7L2/miR-206/Cofilin1 signaling pathway plays an important role in the formation of invadopodia in BCa.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025019
Cancer cells evade immune detection through checkpoint molecules like PD-L1 and PD-L2 which suppress T-cell activation. While PD-L1 is well-studied, the role of PD-L2 remains unclear. Pyruvate kinase M2 (PKM2), a metabolic enzyme, influences immune checkpoint regulation, but its role in PD-L1 and PD-L2 modulation is not well defined. Here, we investigate the role of pyruvate kinase M2 (PKM2) in modulating the immune checkpoint molecules PD-L1 and PD-L2 via GATA3 in cancer cells, with insights from both human and mouse models. We find that PKM2 enhances PD-L1 expression while inhibiting PD-L2, a dual regulatory mechanism that facilitates immune evasion. Knockdown and overexpression experiments revealed GATA3 as a key mediator. PKM2 knockout reduced GATA3 level, leading to decreased PD-L1 and increased PD-L2 expression. Chromatin immunoprecipitation (ChIP)-qPCR demonstrates that GATA3 functions as a direct transcription factor capable of binding to the promoters of PD-L1 and PD-L2. In silico analyses of 81 esophageal squamous cell carcinoma (ESCC) cases from the TCGA database demonstrate that PKM2 mRNA is unrelated to PD-L1 and PD-L2 expression but is negatively correlated with CD8+ T-cell infiltration in ESCC. To further validate these findings, we establish a xenograft model using immune-competent C57/BL6N mice, where knockdown of PKM2 results in significant downregulation of both PD-L1 and PD-L2 expression. Collectively, these findings underscore the divergent roles of PKM2 in regulating immune checkpoint expression in human and mouse cancer models and suggest that targeting the PKM2-GATA3 axis could enhance cancer immunotherapy by fine-tuning PD-L1 and PD-L2 levels.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025154
Mitochondrial dysfunction is closely related to tumor development. Adenine nucleotide translocator 1 (ANT1), which promotes ADP/ATP translocation across the inner mitochondrial membrane, is an important protein involved in mitochondrial function and plays a role in a variety of diseases, including cancers. However, its role in colorectal cancer (CRC) progression remains poorly understood. This study aims to explore the potential role of ANT1 in CRC and its relationship with mitophagy. Through immunohistochemical analysis, we find that ANT1 expression is significantly higher in the tumor tissues of CRC patients than in adjacent normal tissues and that its overexpression is associated with poor prognosis. Further experiments demonstrate that ANT1 knockdown significantly inhibits CRC cell proliferation, migration, and invasion and leads to mitochondrial dysfunction, increased ROS production, and apoptosis by suppressing mitophagy. Mechanistically, ANT1 knockdown downregulates the PINK1/Parkin pathway, thereby inhibiting mitophagy activity. Notably, PINK1 overexpression partially rescues the cellular dysfunction induced by ANT1 knockdown, suggesting a potential role for PINK1 in reversing the suppression of mitophagy. In vivo xenograft models also show that ANT1 knockdown markedly inhibits tumor growth. In conclusion, ANT1 may play a critical role in CRC progression by regulating mitophagy, providing a basis for its potential as a therapeutic target.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024122
The main protease (Mpro) of coronaviruses plays a key role in viral replication, thus serving as a hot target for drug design. PF-00835231 is a promising inhibitor of SARS-CoV-2 Mpro. Here, we report the inhibitory potency of PF-00835231 against SARS-CoV-2 Mpro and seven Mpro mutants (G15S, M49I, Y54C, K90R, P132H, S46F, and V186F) from SARS-CoV-2 variants. The results confirm that PF-00835231 has broad-spectrum inhibition against various coronaviral Mpros. In addition, the crystal structures of SARS-CoV-2 Mpro, SARS-CoV Mpro, MERS-CoV Mpro, and seven SARS-CoV-2 Mpro mutants (G15S, M49I, Y54C, K90R, P132H, S46F, and V186F) in complex with PF-00835231 are solved. A detailed analysis of these structures reveals key determinants essential for inhibition and elucidates the binding modes of different coronaviral Mpros. Given the importance of the main protease for the treatment of coronaviral infection, structural insights into Mpro inhibition by PF-00835231 can accelerate the design of novel antivirals with broad-spectrum efficacy against different human coronaviruses.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024093
Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality worldwide. Increasing evidence suggests that long noncoding RNAs play crucial roles in lung cancer pathogenesis. We previously identified a novel lncRNA, LINC070974, which is associated with tumor cell proliferation. In the present study, we find that knockdown of LINC070974 inhibits cell proliferation, migration and invasion as well as tumor formation both in vitro and in nude mice. LINC070974 silencing also improves cisplatin efficacy in A549/DDP cells. The function of LINC070974 may depend on its interaction with YBX1. Knockdown of LINC070974 reduces the recruitment of YBX1 to the CCND1 promoter and delays tumor progression through its coregulatory genes, which are mainly involved in the p53 signaling pathway. We utilize nebulized inhalation to deliver siRNAs targeting LINC070974 and find that knockdown of LINC070974 significantly prevents tumor metastasis and growth in lung tissues. These findings reveal the role of LINC070974 in lung cancer and suggest a promising therapeutic approach involving siRNA inhalation.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024045
The present study explores the function of FANCA gene, a pivotal member of the Fanconi anaemia (FA) pathway crucial for preserving genomic stability and preventing cancer, particularly in the context of gastric cancer (GC). Using immunohistochemistry, quantitative real-time PCR, and western blot analysis, we evaluate FANCA mRNA and protein expressions in GC cell lines. The relationship between FANCA expression and clinicopathological characteristics is also explored. Various assays, including CCK8, colony formation, wound healing, and Transwell assays, are used to assess functional changes in cells associated with FANCA. Flow cytometry is utilized to evaluate alterations in the cell cycle resulted from FANCA knockdown and overexpression. Our findings show elevated FANCA expression in GC cell lines, with levels correlated with pathologic stage and lymphatic metastasis. FANCA knockdown impedes cell proliferation, migration, and invasion and induces G1/S phase cell cycle arrest. Conversely, FANCA overexpression stimulates cell proliferation, migration, and invasion. In vivo xenograft experiments confirm the promotional role of FANCA in GC tumor progression. Moreover, FANCA overexpression is associated with the activation of cell cycle. Collectively, our results suggest that FANCA drives malignant cell behaviors in GC through the cell cycle pathway, highlighting its potential as a therapeutic target for the treatment of GC.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024155
Hepatocellular carcinoma (HCC), the predominant type of liver cancer, is an aggressive malignancy with limited therapeutic options. In this study, we assess a collection of newly designed gold(I) phosphine complexes. Remarkably, the compound GC002 exhibits the greatest toxicity to HCC cells and outperforms established medications, such as sorafenib and auranofin, in terms of antitumor efficacy. GC002 triggers irreversible necroptosis in HCC cells by increasing the intracellular accumulation of reactive oxygen species (ROS). Mechanistically, GC002 significantly suppresses the activity of thioredoxin reductase (TrxR), which plays a crucial role in regulating redox homeostasis and is often overexpressed in HCC by binding directly to the enzyme. Our in vivo xenograft study confirms that GC002 possesses remarkable antitumor activity against HCC without severe side effects. These findings not only highlight the novel mechanism of controlling necroptosis via TrxR and ROS but also identify GC002 as a promising candidate for the further development of antitumor agents targeting HCC.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024092
Sympathetic activation is a hallmark of heart failure and the underlying mechanism remains elusive. Butyrate is generated by gut microbiota and influences numerous physiological and pathological processes in the host. The present study aims to investigate whether the intestinal metabolite butyrate reduces sympathetic activation in rats with heart failure (HF) and the underlying mechanisms involved. Sprague-Dawley rats (220‒250 g) are anaesthetized with isoflurane, and the left anterior descending artery is ligated to model HF. Then, the rats are treated with or without butyrate sodium (NaB, a donor of butyrate, 10 g/L in water) for 8 weeks. Blood pressure and renal sympathetic nerve activity (RSNA) are recorded to assess sympathetic outflow. Cardiac function is improved (mean ejection fraction, 22.6%±4.8% vs 38.3%±5.3%; P<0.05), and sympathetic activation is decreased (RSNA, 36.3%±7.9% vs 23.9%±7.6%; P<0.05) in HF rats treated with NaB compared with untreated HF rats. The plasma and cerebrospinal fluid levels of norepinephrine are decreased in HF rats treated with NaB. The infusion of N-methyl-D-aspartic acid (NMDA) into the paraventricular nucleus (PVN) of the hypothalamus of HF model rats increases sympathetic nervous activity by upregulating the NMDA receptor. Microglia polarized to the M2 phenotype and inflammation are markedly attenuated in the PVN of HF model rats after NaB administration. In addition, HF model rats treated with NaB exhibit enhanced intestinal barrier function and increased levels of GPR109A, zona occludens-1 and occludin, but decreased levels of lipopolysaccharide-binding protein and zonulin. In conclusion, butyrate attenuates sympathetic activation and improves cardiac function in rats with HF. The improvements in intestinal barrier function, reductions in microglia-mediated inflammation and decreases in NMDA receptor 1 expression in the PVN are all due to the protective effects of NaB.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024054
DPP3, a dipeptidyl peptidase, participates in a variety of pathophysiological processes. DPP3 is upregulated in cancer and might serve as a key factor in the tumorigenesis and progression of various malignancies. However, its specific role and molecular mechanism are still unknown. In this study, the expression of DPP3 in breast cancer tissues is analyzed using TCGA database. Kaplan-Meier survival analysis is performed to estimate the effect of DPP3 on the survival outcomes. To explore the biological function and mechanisms of DPP3 in breast cancer, biochemical and cell biology assays are conducted in vitro. DPP3 expresses at a higher level in breast cancer tissues than that in adjacent tissues in both TCGA database and clinical samples. Patients with high expression of DPP3 have poor survival outcomes. The proliferation and migration abilities of tumor cells with stable DPP3 knockout in breast cancer cell lines are significantly inhibited, and apoptosis is increased in vitro. GSEA analysis shows that DPP3 can affect lipid metabolism and fatty acid synthesis in tumors. Subsequent experiments show that DPP3 could stabilize FASN expression and thus promote fatty acid synthesis in tumor cells. The results of the metabolomic analysis also confirm that DPP3 can affect the content of free fatty acids. This study demonstrates that DPP3 plays a role in the reprogramming of fatty acid metabolism in tumors and is associated with poor prognosis in breast cancer patients. These findings will provide a new therapeutic target for the treatment of breast cancer.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21256
BACKGROUND: Intervertebral disc degeneration is a chronic spinal disease characterized by accelerated apoptosis of nucleus pulposus cells and decomposition of the extracellular matrix, which often leads to low back pain and spinal dysfunction. The molecular mechanism has not been fully understood, hindering the development of precision treatment strategies. Transcriptomics technology can be used to deeply analyze gene expression patterns, and proteomics technology can be used to identify protein function dynamics. The combined application of these two technologies provides an important means to elucidate the pathological mechanism of intervertebral disc degeneration. OBJECTIVE: To review the research progress of transcriptomics and proteomics technologies in intervertebral disc degeneration, and discuss the key roles of these two omics technologies in analyzing molecular mechanisms, screening diagnostic markers, and exploring therapeutic targets. METHODS: A computerized search of relevant literature published from January 1995 to April 2025 in PubMed, Web of Science, CNKI, and Sinomed databases was performed. Chinese search terms were "transcriptomics, ribonucleic acid sequencing, proteomics, intervertebral disc degeneration, nucleus pulposus, annulus fibrosus, cartilage endplate" and English search terms were "transcriptomics, ribonucleic acid sequencing, proteomics, intervertebral disc degeneration, nucleus pulposus, annulus fibrosus, cartilage endplate". After excluding duplicate and irrelevant literature, 24 articles were included for review. RESULTS AND CONCLUSION: Transcriptomics studies have elucidated the dynamic evolution of gene expression in intervertebral disc degeneration, revealing key regulatory networks involved in apoptosis, inflammatory response, and matrix degradation. Proteomics has deeply analyzed the dynamic changes in protein composition in the nucleus pulposus and annulus fibrosus, emphasizing the key functions of matrix metalloproteinases and cytokines in matrix degradation. The combination of the two provides a multi-dimensional perspective on the molecular mechanisms of intervertebral disc degeneration, enhancing the depth of mechanistic research. Comprehensive analysis indicates that the synergistic application of transcriptomics and proteomics shows significant potential in revealing the molecular mechanisms and potential therapeutic targets of intervertebral disc degeneration.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21301
BACKGROUND: The pathogenesis of non-alcoholic fatty liver disease is closely associated with gut microbiota dysbiosis. In recent years, accumulating evidence has indicated that exercise may exert beneficial effects on host metabolic homeostasis by modulating the composition and function of the gut microbiota, thereby playing a positive role in the prevention and treatment of non-alcoholic fatty liver disease. OBJECTIVE: To systematically summarize current research progress on the interplay between gut microbiota and non-alcoholic fatty liver disease, to further elucidate the regulatory effects of exercise on gut microbiota, and to explore in depth the potential mechanisms by which exercise intervention may prevent or ameliorate non-alcoholic fatty liver disease via the "gut–liver axis." METHODS: Search terms included "non-alcoholic fatty liver disease," "gut microbiota," "exercise," "bile acid," "short-chain fatty acid," "lipopolysaccharide," "trimethylamine oxide," and "indole" in Chinese and English, respectively. China National Knowledge Infrastructure (CNKI), and WanFang Database were searched for relevant studies published up to March 2025. A total of 85 core studies were identified based on the inclusion and exclusion criteria. RESULTS AND CONCLUSION: (1) The composition of gut microbiota in patients with non-alcoholic fatty liver disease is significantly abnormal, with increased abundance of pro-inflammatory bacteria (such as Proteobacteria, Escherichia coli, and Streptococcus) and pathogenic bacteria (such as Enterobacteriaceae), while the abundance of anti-inflammatory and homeostatic bacteria (such as Ruminococcus and Faecalibacterium) is decreased. These adverse changes in microbial composition may promote the entry of metabolites into the liver by increasing intestinal permeability, activating inflammatory pathways, and increasing endogenous ethanol production, thereby driving the pathological progression of non-alcoholic fatty liver disease. (2) Modulating gut microbiota through probiotic supplementation or fecal microbiota transplantation can effectively reduce transaminase levels and chronic inflammation in patients with non-alcoholic fatty liver disease, suggesting that gut microbiota may be an important target for the prevention and treatment of non-alcoholic fatty liver disease. (3) Exercise can regulate gut microbiota composition, increase the abundance of beneficial bacteria, reduce the abundance of pro-inflammatory bacteria, and promote the activation of key metabolic pathways, thereby improving host metabolic health. However, current research on the effects of exercise on gut microbiota in patients with non-alcoholic fatty liver disease remains relatively limited, especially the effects and mechanisms of different exercise types, intensities, and durations on gut microbiota and host metabolism are still unclear. (4) Exercise may regulate gut microbiota and increase short-chain fatty acid production to activate G protein-coupled receptors 41/43 and AMP-activated protein kinase pathways, inhibit histone deacetylase activity, thereby reducing hepatic fat accumulation, alleviating liver inflammation, and decreasing insulin resistance; exercise may regulate the gut microbiota-bile acid axis to improve bile acid metabolism, thereby mediating the farnesoid X receptor/G protein-coupled bile acid receptor 5 signaling pathway to prevent and treat non-alcoholic fatty liver disease; exercise may reshape gut microbiota to reduce the abundance of lipopolysaccharide-containing Gram-negative bacteria and improve intestinal barrier function to reduce lipopolysaccharide production and translocation, preventing and treating non-alcoholic fatty liver disease; exercise may regulate gut microbiota composition to enhance the synthesis of indole and its derivatives, while inhibiting the production of ethanol and trimethylamine oxide, thereby enhancing liver metabolic capacity, improving intestinal barrier function, and reducing liver inflammation, playing a positive role in the prevention and treatment of non-alcoholic fatty liver disease.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21280
BACKGROUND: Bushen Tongshi Pills has been proven to delay the progression of collapse in alcoholic femoral head necrosis, but the mechanism is still unclear. OBJECTIVE: To explore the mechanism by which Bushen Tongshi Pills improve osteogenic disorders of alcoholic femoral head necrosis. METHODS: Fifty male Sprague-Dawley rats were randomly divided into a control group, a model group, and low, medium, and high dose groups of Bushen Tongshi Pills, with 10 rats in each group. Except for the control group, other groups were fed with ethanol-containing Lieber-DeCarli liquid diet for 8 weeks to construct alcoholic femoral head necrosis models, while the low, medium, and high dose groups were given 1.05, 2.1, 4.2 g/kg Bushen Tongshi Pills by gavage daily. After 8 weeks, Micro-CT was used to observe the gross morphology of the femoral head, hematoxylin-eosin staining was used to observe the pathological morphology of the femoral head, ELISA was used to detect serum levels of interleukin-1β and interleukin-18, immunohistochemical staining and western blot were used to detect the expression of NOD-like receptor protein 3, Caspase-1, gasdermin D, Runt-related transcription factor 2, osteocalcin, and type I collagen in rat femoral head tissues, and RT-qPCR was used to detect the mRNA expression of these genes. RESULTS AND CONCLUSION: (1) Micro-CT and hematoxylin-eosin staining results showed that the medium and high dose groups of Bushen Tongshi Pills significantly improved bone loss and pathological morphological development in the femoral head of alcoholic femoral head necrosis model rats. (2) ELISA results showed that compared with the control group, serum levels of interleukin-1β and interleukin-18 in the model group were significantly increased (P < 0.05); compared with the model group, serum levels of interleukin-1β and interleukin-18 in all Bushen Tongshi Pills dose groups were significantly decreased (P < 0.05), in a dose-dependent manner, with the most significant decrease in the high dose group (P < 0.05). (3) Immunohistochemistry, western blot, and RT-qPCR results showed that compared with the control group, the protein and mRNA expression of NOD-like receptor protein 3, Caspase-1, and gasdermin D in the femoral head of model rats were significantly upregulated (P < 0.05), while the protein and mRNA expression of osteogenic factors Runt-related transcription factor 2, osteocalcin, and type I collagen were significantly downregulated (P < 0.05); compared with the model group, the protein and mRNA expression of NOD-like receptor protein 3, Caspase-1, and gasdermin D in the femoral head of rats in all Bushen Tongshi Pills dose groups were downregulated in a dose-dependent manner, while the protein and mRNA expression of Runt-related transcription factor 2, osteocalcin, and type I collagen were upregulated in a dose-dependent manner, with the most significant upregulation in the high dose group (P < 0.05). (4) Western blot results showed that compared with the control group, the expression of cleaved-Caspase-1, gasdermin D-N protein, cleaved-Caspase-1/Caspase-1 ratio, and gasdermin D-N/gasdermin D ratio in the femoral head of model rats were significantly upregulated (P < 0.05); compared with the model group, the expression of cleaved-Caspase-1, gasdermin D-N protein, and gasdermin D-N/gasdermin D ratio in all Bushen Tongshi Pills dose groups were significantly downregulated (P < 0.05), and the cleaved-Caspase-1/Caspase-1 ratio in the medium and high dose groups was significantly downregulated (P < 0.05). (5) These results indicate that Bushen Tongshi Pills may inhibit the release of inflammatory factors in alcoholic femoral head necrosis model rats through the NOD-like receptor protein 3/Caspase-1/gasdermin D pyroptosis pathway, promote osteogenic differentiation, and repair necrotic bone tissue.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21417
BACKGROUND: The anterior transpedicular screw fixation system can achieve adequate decompression and strong fixation in a single anterior surgery. However, due to its unique anatomical structure, the risk of screw placement is relatively high. Previous domestic and international scholars have confirmed the feasibility of this approach through anatomical measurements, but there are some drawbacks such as relatively small sample sizes, difficulties in locating the pedicle axis, and limitations in measurement methods. OBJECTIVE: To perform morphological measurements of the adult cervical spine based on imaging to provide anatomical guidance for anterior transpedicular pedicle screw fixation. METHODS: 3D CT scan data of 50 adult cervical vertebrae were imported into the Mimics system for 3D reconstruction. Morphological data were measured, including pedicle axis distance, pedicle width, pedicle height, pedicle horizontal axial angle, pedicle sagittal angle, distance from the entry point in the transverse plane, distance to the entry point in the sagittal plane, axial vertebral length and axial pedicle length. RESULTS AND CONCLUSION: (1) Positioning of nail entry points: C3 and C4 were located on the opposite side of the median sagittal plane of the vertebral body, with distances from the median sagittal line of approximately 2.060 mm and 2.310 mm. C5 could be located on the same side of the median sagittal line as or opposite to the median sagittal line, with an average value of approximately 1.224 mm. C6-C7 were located on the same side of the body, with distances of 1.132 mm and 2.538 mm from the midline; the distance from the upper endplate increased gradually from C3 to C7, with average values ranging from 2.362 to 7.350 mm. (2) Direction of nail entry: the transverse angle increased gradually from C3 to C4 (46.32°-47.36°) and decreased from C5 to C7 (44.03° to 37.80°); the sagittal angle required caudal deviation for C3-C4 (95.75° and 100.93°) and cephalad deviation for C5-C7 (104.38°, 110.34°, and 104.86°). (3) There were no significant differences in entry point location and direction between genders or sides (P > 0.05). For screw selection, except for individual patients with cervical developmental abnormalities, for most subaxial cervical pedicle screws, it is safe and reliable to choose screws at least 30 mm long and 4.0 mm in diameter for males, and at least 28 mm long and 3.5 mm in diameter for females. (4) Morphological measurements confirmed that anterior transpedicular screw fixation of the subaxial cervical spine is feasible, but individualized principles should be followed to formulate personalized fixation plans.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21423
BACKGROUND: Current therapies for musculoskeletal degenerative diseases merely alleviate symptoms with significant adverse effects. As a traditional Chinese medicine, active ingredients of ginseng exhibit protective effects on bone and muscle through multi-target regulation of signaling pathways, and breakthroughs have been made in mechanism research in recent years. OBJECTIVE: To provide a systematical review of the latest molecular mechanisms of active ingredients of ginseng (ginsenosides, polysaccharides, and peptides) in preventing and treating musculoskeletal degenerative diseases via key signaling pathways, providing a basis for targeted drug development. METHODS: A systematic search was performed in multiple databases, including PubMed, Web of Science, Embase, CNKI, Wanfang, and VIP, with the search period from inception to April 2025. Chinese search terms included "musculoskeletal diseases, osteoporosis, osteoarthritis, intervertebral disc herniation, sarcopenia, ginsenosides, ginseng polysaccharides, ginseng peptides, signaling pathways"; English search terms included "musculoskeletal degenerative diseases, osteoporosis, osteoarthritis, intervertebral disc degeneration, sarcopenia, ginsenosides, ginseng polysaccharides, ginseng peptides, signaling pathways". Finally, 75 eligible articles were included. RESULTS AND CONCLUSION: (1) Ginsenosides (e.g., Rg3, Rh4, Rc): target the karyopherin α2-nuclear factor-κB axis to inhibit osteoclast differentiation and reduce bone resorption; activate the sirtuin 1 pathway to enhance mitochondrial biogenesis and delay sarcopenia; regulate Yes-associated protein 1/transcriptional coactivator and p38 mitogen-activated protein kinase to alleviate intervertebral disc degeneration. (2) Polysaccharides: processing techniques affect immunomodulatory activity, inhibiting inflammation via the mitogen-activated protein kinase/nuclear factor-κB pathway. (3) Peptides: activate the NAD+/sirtuin 1/peroxisome proliferator-activated receptor γ coactivator 1α axis to improve mitochondrial function. These findings suggest that ginseng exerts synergistic regulation of bone metabolism balance, inhibition of cartilage degradation, and delay of muscle aging through a "multi-component, multi-target" mechanism, but challenges remain regarding low bioavailability and lack of large-scale clinical trials for translation.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21507
BACKGROUND: As a common condition of the musculoskeletal system, triangular fibrocartilage complex injury necessitates effective treatment strategies to improve patients' quality of life and functional recovery. However, there is a lack of systematic integration of global research trends and core hotspots. OBJECTIVE: To conduct a bibliometric analysis of the literature on triangular fibrocartilage complex injury treatment, aiming to explore the current research status, hotspots, and trends in this field. METHODS: Relevant literature in the field of triangular fibrocartilage complex injury treatment published between 2001 and 2024 was retrieved from the Web of Science Core Collection. Citespace software was utilized for visual analysis of countries, authors, institutions, co-cited references, and keywords. RESULTS AND CONCLUSION: (1) After screening, 352 articles were included, with an increasing trend in the annual publication volume. The United States was the leading contributing country, and Mayo Clinic (USA) was the most prolific institution. (2) The top five most frequent keywords were triangular fibrocartilage complex, wrist, distal radioulnar joint, anatomy, and tear, with 'anatomy' having the highest centrality. (3) Temporal analysis revealed that early research focused on anatomical foundations and diagnostic methods for wrist injuries; from 2010 onward, the focus shifted to complications and long-term outcomes after arthroscopic surgery. In recent years, research has increasingly emphasized precision, standardization, and interdisciplinary integration. (4) Research on triangular fibrocartilage complex injury treatment has entered an innovative phase of multidisciplinary convergence, with vigorous development in areas such as precision anatomy, intelligent diagnosis and treatment, and biomaterials.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21489
BACKGROUND: The pathological process of Alzheimer's disease is closely related to β-amyloid/Tau deposition and cerebrovascular dysfunction, and existing therapies are difficult to effectively intervene. In recent years, research on glucagon-like peptide-1 receptor agonists for the treatment of Alzheimer's disease has gained increasing popularity. As a novel dual agonist of the glucagon-like peptide-1 receptor and glucagon receptor, the therapeutic mechanism of Mazdutide in Alzheimer’s disease remains to be elucidated.
OBJECTIVE: To systematically explore the mechanism by which Mazdutide improves cognitive function in Alzheimer's disease using network pharmacology and experimental validation, and to identify potential core molecular targets, providing a theoretical basis for new therapeutic strategies.
METHODS: A multi-omics integration strategy was employed. Co-localization targets of Alzheimer's disease and Mazdutide were screened using DisGeNET and SEA databases. A protein-protein interaction network was constructed via STRING, and hub genes were identified by Cytoscape topological analysis. In APP/PS1/Tau triple transgenic Alzheimer's disease mice, Mazdutide (30 nmol/kg) was administered intraperitoneally. Cognitive function was assessed using Morris water maze, novel object recognition, and Y maze tests. Western blot was used to detect hippocampal β-amyloid (6E10), p-Tau181, and endothelin receptor A expression.
RESULTS AND CONCLUSION: ① Fifty-two co-localized targets were identified, with endothelin receptor A (EDNRA) as the top hub gene. ② Mazdutide significantly improved cognition in 3xTg mice: shortened spatial memory latency, increased novel object recognition index, increased spontaneous alternation in Y maze, and reduced hippocampal pathological burden, with decreased p-Tau181 and EDNRA overexpression. ③ Gene Ontology and KEGG enrichment revealed core pathways: biological processes included blood pressure regulation, amine transport regulation, and amine transport; cellular components included symmetric synapses, sperm head, and pseudopodia; molecular functions included G protein-coupled peptide receptor activity, peptide receptor activity, and neuropeptide receptor activity. KEGG pathways included neuroactive ligand-receptor interaction, calcium signaling, and hormone signaling. ④ This study confirms that Mazdutide may improve cognitive deficits in Alzheimer's disease by targeting and inhibiting EDNRA overexpression, reducing neuronal pathological damage, and acting through multiple dimensions. The discovery of EDNRA provides a novel therapeutic target for Alzheimer's disease.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21478
BACKGROUND: The polarization state of macrophages is closely related to the occurrence and development of knee osteoarthritis. Luteolin can regulate the nuclear transcription factor κB (NF-κB) signaling pathway to affect the polarization process of macrophages, but the specific mechanism remains unclear. OBJECTIVE: To investigate the mechanism of luteolin in the treatment of knee osteoarthritis. METHODS: (1) Intersectional targets of luteolin, macrophage polarization, and knee osteoarthritis were screened by network pharmacology, a protein-protein interaction network was constructed, and GO, KEGG enrichment analysis and molecular docking were performed. (2) After knocking out NF-κB p65, RAW264.7 cells were induced to polarize to M1 and M2 types, and flow cytometry was used to detect cell polarization tendency. Different concentrations of luteolin were used to intervene in RAW264.7 cells, and the appropriate concentration of luteolin was screened by CCK-8 method for subsequent experiments. RAW264.7 cells were induced to polarize to M1 and M2 types, and luteolin was added for intervention, and flow cytometry was used to detect cell polarization tendency. RAW264.7 cells were induced to polarize to M1 type, and luteolin was added for intervention. ELISA was used to detect the levels of tumor necrosis factor α, interleukin-6, and interleukin-10 in the cell supernatant, immunofluorescence staining was used to detect the nuclear translocation of NF-κB p65/p50, and western blot was used to detect the expression and phosphorylation of NF-κB p65 and NF-κB inhibitor protein α. RESULTS AND CONCLUSION: (1) A total of 137 intersection genes were screened, of which 135 target genes were involved in the construction of the protein-protein interaction network. GO and KEGG analysis showed that the NF-κB complex was enriched in the protein-protein interaction network. Molecular docking showed that luteolin docked well with NF-κB inhibitor protein α and NF-κB p50/p65 proteins. (2) After NF-κB p65 knockout, the polarization tendency of RAW264.7 cells to M1 type was inhibited, and spontaneous M2 polarization tendency appeared. Luteolin intervention could inhibit the polarization of RAW264.7 cells to M1 type and promote the polarization to M2 type. During the induction of RAW264.7 cells to M1 polarization, luteolin intervention could inhibit the release of pro-inflammatory factors, promote the release of anti-inflammatory factors, inhibit the nuclear translocation of NF-κB p65/p50, promote the expression of NF-κB inhibitor protein α and inhibit its phosphorylation, and inhibit the expression and phosphorylation of NF-κB p65. The results indicate that luteolin can regulate macrophage polarization through the NF-κB signaling pathway and has a potential therapeutic effect on knee osteoarthritis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21700
BACKGROUND: Some patients report dissatisfaction after total knee arthroplasty due to persistent issues like limited mobility, joint instability, and residual pain. To address these challenges, various osteotomy methods and alignment strategies have been proposed and clinically implemented. However, the optimal approach remains debated, with no clear consensus. OBJECTIVE: To comprehensively analyze global trends in alignment techniques for total knee arthroplasty and identify research hotspots through bibliometric and visual analysis, offering evidence-based insights to guide clinical decision-making. METHODS: We retrieved publications (articles and reviews) on total knee arthroplasty alignment techniques published between January 1, 2015 and December 31, 2024, from the Web of Science database (https://www.webofscience.com, developed by the Institute for Scientific Information, USA). Data on countries, institutions, publication years, authors, journals, mean citation rates, H-index, titles, keywords, and the top 25 most-cited articles were extracted and analyzed. VOSviewer and Citespace software were used to analyze keyword co-occurrence and predict research hotspots. RESULTS AND CONCLUSION: A total of 866 publications were identified, including 792 articles and 74 reviews. The United States had the highest number of publications, with the University of California being the most prolific institution and Howell, Stephen M being the most prolific author. The journal 'Knee Surgery Sports Traumatology Arthroscopy' had the highest publication and citation counts. Common keywords included total knee arthroplasty, alignment technique, arthroplasty, clinical outcomes, and survival rate. Keyword and reference burst detection indicated that kinematic alignment and robotic navigation are future research hotspots. The number of publications on total knee arthroplasty alignment techniques is increasing annually. The United States is currently the leading contributor, followed by the United Kingdom. Increasingly precise tools, such as computer-assisted navigation and patient-specific instrumentation, are being developed and applied to achieve alignment goals, but no optimal solution has yet been established. Future research frontiers may include 'robot-assisted navigation', 'patient satisfaction', 'long-term survival', 'indication range', and 'patient-specific alignment'.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21573
BACKGROUND: Patients undergoing mandibular third molar extraction show significantly higher dental anxiety levels than those receiving other dental procedures. Preoperative anxiety not only prolongs operation time but also worsens postoperative complications. OBJECTIVE: To investigate the relationship between preoperative dental anxiety and the time of mandibular third molar extraction. METHODS: A cross-sectional observational study was designed for the patient's extraction of mandibular third molar in the outpatient clinic of Department of Stomatology of Aerospace Center Hospital. The Modified Dental Anxiety Scale for Third Molar Surgery (MDAS-TMS) was used to evaluate the preoperative anxiety status. Demographic characteristics, surgical and radiographic features of the patients were recorded. The main outcome indicator was the duration of surgery. Multivariable linear regression models were developed to analyze their influences, and a restricted cubic spline function was used to evaluate their potential nonlinear associations. RESULTS AND CONCLUSION: A total of 216 patients (71 men and 145 women) were included in the study. The mean DAS-TMS score was 10.4, 32.4% were mildly anxious, and 8.8% were highly anxious. For each 1-unit increase in the DAS-TMS after multivariate adjustment, operation time increased by 1.1 minutes (β=1.10, P < 0.001, 95%CI: 0.76-1.49). Restricted cubic spline analysis revealed evidence of a nonlinear association (P for nonlinearity=0.013). The results indicate that preoperative dental anxiety has a significant impact on the duration of mandibular third molar extraction.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026045
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, necessitating the discovery of novel therapeutic agents. Here, we report a natural small molecule, 2-dihydroailanthone (2-DAIL), as a promising candidate for CRC treatment. First, our results demonstrate that 2-DAIL exhibits significant anti-CRC activity in vitro and in vivo. Then, we find that 2-DAIL directly binds to integrin alpha-3 (ITGA3) revealed by stable isotope labeling by amino acids in cell culture coupled with thermal proteome profiling (SILAC-TPP). Additionally, the RNA sequencing data obtained from CRC cells and tumor tissues suggest that 2-DAIL blocks the PI3K/AKT signaling pathway mediated by ITGA3 inhibition. Collectively, 2-DAIL exerts its anti-CRC effects, at least partially, by binding to and inhibiting the function of ITGA3, thereby blocking the activation of the PI3K/AKT signaling pathway, which leads to CRC cell growth inhibition. Our study provides a promising drug candidate for the treatment of CRC and suggests the potential of 2-DAIL in treating other diseases linked to ITGA3 dysfunction.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025181
Colorectal cancer (CRC) is one of the most prevalent and lethal cancers worldwide and is characterized by uncontrolled cell invasion, migration, and proliferation. The progression of CRC is driven by genetic mutations and alterations in key signaling pathways. This study investigates the role of GPSM2 and ZNF263 implicated in CRC progression. The GPSM2 gene, which regulates G protein signaling pathways, plays a vital role in cell movement and growth, contributing to the metastatic potential of cancer cells. The ZNF263 gene, a zinc finger protein involved in gene expression regulation, is also linked to CRC progression, with its dysregulation affecting the cell cycle, apoptosis, and migration. In particular, this study explores how ZNF263 acts as a transcription factor, modulating the expression of GPSM2 to increase CRC cell invasion, migration, and proliferation. This study confirms that ZNF263 activates the cell cycle pathway in a GPSM2-dependent manner, driving the aggressive behavior of CRC cells. Bioinformatics analysis using the GEO database further supports these findings, identifying key genetic alterations in CRC. These insights provide a deeper understanding of the molecular mechanisms underlying CRC progression and highlight the potential of ZNF263 and GPSM2 as therapeutic targets for intervention. This study underscores the importance of early detection and exploration of targeted therapies to improve CRC patient outcomes.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026035
Pulmonary fibrosis (PF) is a life-threatening interstitial lung disease characterized by scarring and inflammation in lung tissues. Aberrant activation of the JAK/STAT and NF-κB signaling pathways is critical in initiating and sustaining the inflammatory processes that drive fibrotic progression. In this study, we identify a novel small-molecule compound, T4015, a 4-indolyl-2-phenylaminopyrimidine derivative, as a dual-pathway inhibitor targeting both JAK/STAT and NF-κB signaling. Dual-luciferase reporter assays demonstrate the potent inhibitory activity of T4015 against these pathways. T4015 effectively suppresses the phosphorylation of STAT3, JAK1, and TYK2 induced by IL-6 and IFN-β, while suppressing LPS-induced NF-κB activation in macrophages. Transcriptome sequencing and pathway enrichment analyses further confirm that T4015 downregulates multiple inflammation-related signaling cascades, including the JAK/STAT, NF-κB, TNF, IL-17, and Toll-like receptor pathways. In a mouse model of bleomycin-induced PF, T4015 treatment significantly improves survival, attenuates collagen deposition, and reduces the expression of pro-inflammatory and profibrotic markers such as IL-6, CCL2, and COL1. Molecular docking and target prediction analyses suggest that T4015 exhibits strong binding affinity for multiple kinases within the JAK/STAT and NF-κB networks, including JAK1, TYK2, JAK2, JAK3, RIPK1, IRAK1/4, TAB1, and ZAP70. Collectively, these results highlight T4015 as a promising therapeutic candidate for PF through its simultaneous inhibition of the JAK/STAT and NF-κB signaling pathways.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025019
Cancer cells evade immune detection through checkpoint molecules PD-L1 and PD-L2, which suppress T-cell activation. While PD-L1 is well-studied, the role of PD-L2 remains unclear. Pyruvate kinase M2 (PKM2), a metabolic enzyme, influences immune checkpoint regulation, but its role in PD-L1 and PD-L2 modulation is not well defined. Here, we investigate the role of PKM2 in modulating PD-L1 and PD-L2 via GATA3 in cancer cells, with insights from both human and mouse models. We find that PKM2 enhances PD-L1 expression while inhibiting PD-L2, a dual regulatory mechanism that facilitates immune evasion. Knockdown and overexpression experiments revealed GATA3 as a key mediator. PKM2 knockout reduced GATA3 level, leading to decreased PD-L1 and increased PD-L2 expression. Chromatin immunoprecipitation (ChIP)-qPCR demonstrates that GATA3 functions as a direct transcription factor capable of binding to the promoters of PD-L1 and PD-L2. In silico analyses of 81 esophageal squamous cell carcinoma (ESCC) cases from the TCGA database demonstrate that PKM2 mRNA is unrelated to PD-L1 and PD-L2 expression but is negatively correlated with CD8+ T-cell infiltration in ESCC. To further validate these findings, we establish a xenograft model using immune-competent C57/BL6N mice, where knockdown of PKM2 results in significant downregulation of both PD-L1 and PD-L2 expression. Collectively, these findings underscore the divergent roles of PKM2 in regulating immune checkpoint expression in human and mouse cancer models and suggest that targeting the PKM2-GATA3 axis could enhance cancer immunotherapy by fine-tuning PD-L1 and PD-L2 levels.