Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04896-8
Background The development of vascular calcification (VC) in diabetes is closely related to the endothelial-to-mesenchymal transition (EndMT). We found that microRNA-32-5p (miR-32) was elevated in the plasma of calcification patients. However, it is unclear whether miR-32 mediates the function of bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) in type 2 diabetes (T2D) VC. Methods BMSC-EVs were characterized by TEM, NTA, Western blotting, and confocal microscopy. Alizarin Red and ALP staining assessed the severity of VC. qRT-PCR and Western blotting evaluated the expression of BMP2, RUNX2, GPX4, SLC7A11, VE-cadherin, and N-cadherin, while immunofluorescence was used for detecting VE-cadherin and N-cadherin. In vivo validation was performed using miR-32–/– and ApoE–/– mice. RNA sequencing (RNA-seq) and bioinformatics analysis was conducted to explore underlying mechanisms. Results We demonstrated that BMSC-EVs attenuate VC in endothelial cells (ECs) and inhibit EndMT. In vivo, histological analysis showed that treatment with BMSC-EVs significantly reduced the severity of VC associated with T2D. Notably, knockout of miR-32 further enhanced the inhibitory effect of BMSC-EVs on VC. Mechanistically, transcriptomic and functional analyses suggest that the protective effect of BMSC-EVs on VC is associated with regulation of the MAPK/FoxO signaling pathway, potentially mediated by modulation of ferroptosis. Conclusion These findings demonstrate that BMSC-EVs attenuate T2D-associated VC, partially through miR-32-mediated suppression of EC ferroptosis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04173-0
Background Cavernous nerve injury-induced erectile dysfunction (CNI-ED) is a common complication after radical prostatectomy. Conventional treatment approaches have had little success in treating the severe cavernous fibrosis which is a consequence of CNI-ED. Methods Pre-treatment of adipose-derived stem cells with melatonin allows for the extraction of active exosomes (MT-hASC-EVs) from the conditioned medium. The therapeutic effects of MT-hASC-EVs were assessed in a rat model of CNI-ED, and the anti-fibrotic properties were evaluated. MicroRNA sequencing was used to identify specific microRNAs highly expressed in MT-hASC-EVs, and differential microRNAs were screened for regulatory pathways through target gene enrichment analysis. Finally, the conclusions from bioinformatics analysis were validated through in vitro experiments. Results Intracavernous injection of MT-hASC-EVs significantly restored erectile function and reduced the extent of corpus cavernosum fibrosis in the CNI-ED rat model. MT-hASC-EVs promoted the proliferation and anti-apoptotic effects of corpus cavernosum smooth muscle cells (CCSMCs) in vitro. Mechanistically, MT-hASC-EVs inhibit fibrosis by delivering miR-145-5p, which targets TGF-β2/Smad3 axis. Conclusions MT-hASCs-EVs can inhibit cavernous fibrosis and improve erectile function in a rat model of CNI-ED by targeting the miR-145-5p/TGF-β/Smad axis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04637-3
Background Human umbilical cord mesenchymal stem cells (hUMSCs) are considered an effective prospect for treating TBI, but they tend to accumulate in the lungs after intravenous injection, hindering further clinical translation. Brahma-related gene 1(BRG1) can be influenced by estrogen to regulate adhesion, and ourprevious studies have found that the expression of BRG1 in lungs increases after TBI. However, the relationship between BRG1, estrogen, TBI, and stem cell lung aggregation is not clear. Methods By regulating the expression levels of BRG1 in vascular endothelial cells and hUMSCs, Western Blot and immunohistochemistry were used to explore its changes in adhesion and possible mechanisms; used in vivo bioluminescenece imaging analysis, real-time tracking the distribution of stem cells after transplantation; and therapeutic drug E2 is introduced to observe the effect of changes in BRG1 expression on the aggregation of hUMSCs in the lungs of model animals, as well as the therapeutic effect of E2-pretreated hUMSCs on inflammation after TBI. Results After TBI, the retention of hUMSCs in the lungs was higher in the TBI groups than in the Sham groups, and the level of BRG1 in lung was higher in the TBI groups than in the Sham groups; the expression of BRG1 in HUVECs, HPAECs, and hUMSCs treated with TNF-α and LPS were higher than those in the control groups, showing dose- and time-dependent effects. E2 can inhibit the expression of BRG1 and adhesion proteins; after intervention with estrogen receptor inhibitor (ICI 182780) and NF-κ B inhibitor SC75741, BRG1 expression increased and adhesion protein decreased; E2-pretreated MSCs can reduce pulmonary retention, and has no adverse effects on the inflammatory response for TBI.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03835-9
Background Facial infiltrating lipomatosis is characterized by excessive growth of adipose tissue. Its etiology is associated with somatic phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) variants, but the specific mechanisms are not yet fully understood. Methods We collected facial adipose tissue from both FIL patients and non-FIL individuals, isolated the stromal vascular fraction (SVF) and performed single-cell transcriptome sequencing on these samples. Results We mapped out the cellular landscape within the SVF, with a specific focus on a deeper analysis of fibro-adipogenic precursor cells (FAPs). Our analysis revealed that FAPs from FIL patients (FIL-FAPs) significantly overexpressed FK506 binding protein 51 (FKBP5) compared to FAPs from individuals without FIL. Further experiments indicated that FKBP5 is regulated by the PI3K-AKT signaling pathway. The overactivation of this pathway led to an increase in FKBP5 expression. In vitro experiments demonstrated that FKBP5 promoted adipogenic differentiation of FAPs, a process that could be hindered by FKBP5 knockdown or inhibition. Additionally, in vivo assessments confirmed FKBP5’s role in adipogenesis. Conclusions These insights into the pathogenesis of FIL underscore FKBP5 as a promising target for developing non-surgical interventions to manage the excessive adipose tissue growth in FIL.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024049
Chronic renal failure (CRF) is a severe syndrome affecting the urinary system for which there are no effective therapeutics. In this study, we investigate the effects and mechanisms of aminophylline in preventing CRF development. A rat model of chronic renal failure is established by 5/6 nephrectomy. The levels of serum creatinine (SCR), urinary protein (UPR), and blood urea nitrogen (BUN) are detected by ELISA. Histological evaluations of renal tissues are performed by H&E, Masson staining, and PAS staining. Functional protein expression is detected by western blot analysis or immunofluorescence microscopy. Glomerular cell apoptosis is determined using the TUNEL method. Results show that Aminophylline significantly reduces the levels of SCR, UPR, and BUN in the CRF model rats. Histological analyses show that aminophylline effectively alleviates renal tissue injuries in CRF rats. The protein expression levels of nephrin, podocin, SIRT1, p-AMPK, and p-ULK1 are greatly increased, while p-mTOR protein expression is markedly decreased by aminophylline treatment. Additionally, the protein level of LC3B in CRF rats is significantly increased by aminophylline. Moreover, aminophylline alleviates apoptosis in the glomerular tissues of CRF rats. Furthermore, resveratrol promotes SIRT1, p-AMPK, and p-ULK1 protein expressions and reduces p-mTOR and LC3B protein expressions in CRF rats. Selisistat (a SIRT1 inhibitor) mitigates the changes in SIRT1, p-AMPK, p-ULK1, p-mTOR, and LC3B expressions induced by aminophylline. Finally, RAPA alleviates renal injury and apoptosis in CRF rats, and 3-MA eliminates the aminophylline-induced inhibition of renal injury and apoptosis in CRF rats. Aminophylline suppresses chronic renal failure progression by modulating the SIRT1/AMPK/mTOR-mediated autophagy process.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024024
Myocardial ischemia/reperfusion (I/R) injury is a classic type of cardiovascular disease characterized by injury to cardiomyocytes leading to different types of cell death. The degree of irreversible myocardial damage is closely related to age, and ferroptosis is involved in cardiomyocyte damage. However, the mechanisms underlying ferroptosis regulation in aging myocardial I/R injury are still unclear. The present study aims to explore the underlying mechanism of piRNA regulation in ferroptosis. Using left anterior descending coronary artery ligation in an aging rat model and a D-galactose-induced rat cardiomyocyte line (H9C2) to construct an aging cardiomyocyte model, we investigate whether ferroptosis occurs after reperfusion injury in vitro and in vivo. This study focuses on the upregulation of piR-000699 after hypoxia/reoxygenation treatment in aging cardiomyocytes by observing hypoxia/reoxygenation (H/R) injury indicators and ferroptosis-related indicators and clarifying the role of piR-000699 in H/R injury caused by ferroptosis in aging cardiomyocytes. Bioinformatics analysis reveals that SLC39A14 is a gene that binds to piR-000699. Our data show that ferroptosis plays an important role in I/R injury both in vivo and in vitro. Furthermore, the results show the potential role of piR-000699 in regulating SLC39A14 in ferroptosis in aging cardiomyocytes under hypoxia/reoxygenation conditions. Together, our results reveal that the mechanism by which piR-000699 binds to SLC39A14 regulates ferroptosis in aging myocardial I/R injury.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024222
We aim to identify molecular clusters related to O-GlcNAcylation and establish a novel scoring system for predicting prognosis and immunotherapy efficacy in patients with gastric cancer (GC). The transcriptomic and clinical data are obtained from XENA-UCSC and GEO databases. The O-GlcNAcylation-related genes are obtained from the GSEA database. Consensus clustering analysis is employed to identify O-GlcNAcylation-related molecular clusters, and principal component analysis (PCA) is utilized to develop a novel prognostic scoring system for predicting GC outcomes and immunotherapy efficacy. The prognostic accuracy of the scoring system is assessed across five real-world cohorts. The biological function of actin alpha 2, smooth muscle (ACTA2) in GC is determined through experimental verification. Using 34 O-GlcNAcylation-related genes associated with prognosis in GC patients, these individuals are divided into two distinct subgroups characterized by different outcomes, tumor microenvironment profiles, and clinical case characteristics. The DEGs between the two subgroups are subsequently used to further divide the GC patients into two subgroups by consensus cluster analysis. PCA is used to construct a prognostic scoring system, which reveal that patients in the low-score subgroup have a better prognosis and greater benefit from immunotherapy. The accuracy of the scoring system is confirmed through validation in a cohort of patients receiving immunotherapy in the real world. ACTA2 promotes proliferation and inhibits apoptosis in GC cells. These findings suggest that we successfully establish molecular clusters associated with O-GlcNAcylation and develop a scoring system that demonstrates strong performance in predicting the prognosis of patients with GC and the effect of immunotherapy interventions.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024181
MicroRNAs (miRNAs) have emerged as essential regulators that play important roles in the development of multiple systems. Recent studies have identified significant roles for miRNAs in the progression of cardiac hypertrophy. This study aims to investigate the effects of miR-133b-3p on angiotensin II (Ang II)-induced cardiac hypertrophy and apoptosis, as well as explore its underlying mechanisms. Our experimental results reveal that miR-133b-3p expression is significantly decreased in both animal and cell models of cardiac hypertrophy induced by Ang II. Overexpression of miR-133b-3p reverses the hypertrophic manifestations and apoptosis induced by Ang II. Through bioinformatics analysis and dual-luciferase reporter assays, CDIP1 (cell death inducing p53 target 1) is identified as a direct target of miR-133b-3p, and the overexpression of miR-133b-3p reduces CDIP1 expression. Additionally, CDIP1 silencing suppresses cardiomyocyte hypertrophy and apoptosis induced by Ang II. In summary, these results suggest that miR-133b-3p may serve as a potential diagnostic marker for cardiac hypertrophy and that the upregulation of miR-133b-3p inhibits cardiac hypertrophy by targeting CDIP1.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024178
Endometriosis (EMS) is a benign gynecological disease characterized by the growth of endometrial tissue outside the uterine cavity. Evidence shows that the survival of patients with ectopic endometrial implants is associated with a dysregulated immune microenvironment. CD4+ T cells can regulate EMS through diverse cytokines, the inflammatory response, and angiogenesis. CCR5+CD4+ T cells exhibit increased cellular immunogenicity and play a role in infectious diseases, host defense, and cancer progression. However, the specific mechanisms of CCR5+CD4+ T cells in EMS remain unknown. In the present study, flow cytometry and RNA-seq are utilized to assess the proportions and features of CCR5+CD4+ T cells in EMS patients, RT-PCR and ELISA are used to assess the production of CCL5 by ectopic endometrial stromal cells (ecESCs). Two EMS models are established through C57B6 wild-type and CCL5‒/‒ mice and utilized to explore the in vivo effects of CCR5+CD4+ T cells on ectopic lesions. Compared with CCR5‒CD4+ T cells, CCR5+CD4+ T cells display a more activated and cytotoxic phenotype. Diminished CCR5+CD4+ T cells and their impaired ability to produce IFN-γ are observed in the ectopic lesions of EMS patients and in murine EMS models. Impaired production of CCL5 has been detected in human ecESCs. Moreover, endometria stripped from CCL5‒/‒ mice are more likely to generate ectopic lesions in the peritoneum of recipient mice. These findings demonstrate that the attenuated recruitment of CCR5+CD4+ T cells in ectopic lesions caused by decreased production of CCL5 in ecESCs may facilitate the progression of EMS.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025221
Hypertension is commonly accompanied by endothelial dysfunction, characterized by an imbalance between vasodilatation and constriction, increased levels of the proinflammatory factors interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1), and decreased nitric oxide (NO) bioavailability. Using an angiotensin II (Ang II)-induced endothelial dysfunction model, we show that treatment with the hydrogen sulfide (H₂S) donor GYY4137 significantly reverses Ang II-induced damage. GYY4137 restores sirtuin 6 (SIRT6) expression, suppresses inflammation, and improves vasodilatory function. Furthermore, endothelial-specific cystathionine-γ-lyase (CSE)-deficient mice exhibit inflammation and endothelial dysfunction in blood vessels, which is reversed by H₂S supplementation. Critically, SIRT6 inhibitors block the protective effects of H₂S in the endothelium. This study demonstrates that H₂S protects vascular endothelial function by activating the SIRT6 anti-inflammatory pathway.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024074
Alcoholic liver disease (ALD) poses a significant health challenge, so comprehensive research efforts to improve our understanding and treatment strategies are needed. However, the development of effective treatments is hindered by the limitation of existing liver disease models. Liver organoids, characterized by their cellular complexity and three-dimensional (3D) tissue structure closely resembling the human liver, hold promise as ideal models for liver disease research. In this study, we use a meticulously designed protocol involving the differentiation of human induced pluripotent stem cells (hiPSCs) into liver organoids. This process incorporates a precise combination of cytokines and small molecule compounds within a 3D culture system to guide the differentiation process. Subsequently, these differentiated liver organoids are subject to ethanol treatment to induce ALD, thus establishing a disease model. A rigorous assessment through a series of experiments reveals that this model partially recapitulates key pathological features observed in clinical ALD, including cellular mitochondrial damage, elevated cellular reactive oxygen species (ROS) levels, fatty liver, and hepatocyte necrosis. In addition, this model offers potential use in screening drugs for ALD treatment. Overall, the liver organoid model of ALD, which is derived from hiPSC differentiation, has emerged as an invaluable platform for advancing our understanding and management of ALD in clinical settings.
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.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04896-8
Vascular calcification (VC) in type 2 diabetes (T2D) is driven by endothelial-to-mesenchymal transition (EndMT), yet effective therapies remain elusive. Elevated plasma microRNA-32-5p (miR-32) correlates with calcification, but its role in bone marrow mesenchymal stem cell-derived extracellular vesicle (BMSC-EV) therapy is undefined. We characterized BMSC-EVs by TEM, NTA, Western blotting, and confocal microscopy. Alizarin Red and ALP staining quantified VC severity. qRT-PCR and Western blotting assessed BMP2, RUNX2, GPX4, SLC7A11, VE-cadherin, and N-cadherin; immunofluorescence localized VE-cadherin and N-cadherin. In vivo validation used miR-32–/– and ApoE–/– mice. RNA sequencing and bioinformatics explored mechanisms. BMSC-EVs attenuated VC in endothelial cells (ECs) and inhibited EndMT. In vivo, BMSC-EV treatment significantly reduced T2D-associated VC severity. Notably, miR-32 knockout further enhanced the inhibitory effect of BMSC-EVs on VC. Transcriptomic and functional analyses linked the protective effect to MAPK/FoxO signaling modulation, potentially via ferroptosis regulation. These findings demonstrate that BMSC-EVs attenuate T2D-associated VC, partially through miR-32-mediated suppression of EC ferroptosis, providing a mechanistic foundation for EV-based therapeutics.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21312
BACKGROUND: Although physcion has been shown to have protective effects against osteoporosis, the exact mechanism is not fully understood. OBJECTIVE: Through multidimensional analysis of the regulatory effect of physcion on the AKT signaling pathway, the molecular mechanism of its regulation on osteoclast induced differentiation and osteogenic function induced differentiation is revealed. METHODS: (1) RAW264.7 cells and C3H10T1/2 cells were cultured in vitro and subsequently exposed to 0, 10, 20, 30, 40, 50, and 60 µmol/L physcion, respectively. The cytotoxicity of physcion was detected by cell counting kit-8 assay. (2) RAW264.7 cells and C3H10T1/2 cells were treated with different concentrations (0, 20, 40 µmol/L) of physcion during osteoclast and osteoblast differentiation, respectively. Differentiation ability was assessed by qPCR, Western Blot, and alkaline phosphatase staining. (3) Network pharmacology was used to analyze the regulation of physcion on osteoclast differentiation and related signaling pathways, and molecular docking was performed for target proteins. (4) Western Blot was used to verify the phosphorylation level of AKT in the downstream target signaling pathway AKT axis regulated by physcion. RESULTS AND CONCLUSION: (1) At concentrations of 0-60 µmol/L, cell viability in all groups was greater than 90%, indicating no significant cytotoxicity. (2) Physcion significantly inhibited the expression of osteoclast differentiation-related genes, with Acp5, CTSK, DC-STAMP, and Nfatc1 showing downregulation, but had no significant effect on osteoblast differentiation-related genes COL1A1, Runx2, OSX expression or alkaline phosphatase staining intensity. (3) Network pharmacology and molecular docking suggested that physcion affects osteoclast differentiation and regulates the PI3K-AKT pathway, with a binding energy of -10.72 kJ/mol to AKT1, indicating strong binding activity. (4) During osteoclast differentiation, the p-AKT/AKT ratio in RAW264.7 cells increased (n=3, P=0.0063), while physcion decreased this ratio. These findings indicate that physcion inhibits osteoclast differentiation by regulating the AKT signaling pathway, thereby modulating bone homeostasis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21403
BACKGROUND: Lycium barbarum polysaccharide is a natural active ingredient with potential to lower blood glucose and improve diabetes-related symptoms. However, from the perspective of gut microbiota, the underlying factors for the effects of Lycium barbarum polysaccharide on glycolipid abnormalities have not been fully elucidated. OBJECTIVE: To investigate the effect of Lycium barbarum polysaccharide on type 2 diabetes mellitus and its related mechanism. METHODS: Six male Sprague-Dawley rats aged 8 weeks were randomly selected from 18 rats to form a blank control group. The remaining 12 rats were fed a high-sugar, high-fat diet for 8 weeks and then received a single tail vein injection of 1% streptozotocin to establish a type 2 diabetes model. After successful modeling, the rat models were then randomly divided into a model control group (n=6) and a Lycium barbarum polysaccharide group (n=6). Rats in the Lycium barbarum polysaccharide group were administered Lycium barbarum polysaccharide solution via gavage at a dose of 200 mg/(kg·d), 2 mL per dose, once daily, for 12 consecutive weeks. After the intervention, rat serum and feces were collected. 16S rDNA sequencing was used to analyze gut microbiota, and alpha diversity, beta diversity, and principal component analysis were used to characterize microbial abundance and structure. Liquid chromatography-mass spectrometry was used to detect short-chain fatty acid levels. ELISA was used to detect glycolipid metabolism, insulin resistance, and inflammatory response indicators. RESULTS AND CONCLUSION: Compared with the model control group, the Lycium barbarum polysaccharide group had increased high-density lipoprotein cholesterol levels and decreased total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels, indicating that Lycium barbarum polysaccharide improved lipid accumulation and inhibited weight loss in type 2 diabetic rats. Compared with the model control group, the Lycium barbarum polysaccharide group had decreased levels of interleukin-6, tumor necrosis factor-alpha, and blood glucose, and increased levels of insulin and glucagon-like peptide-1, indicating that Lycium barbarum polysaccharide effectively inhibited inflammatory response and insulin resistance. Lycium barbarum polysaccharide significantly improved the composition of gut microbiota, increasing the abundance of Lachnospiraceae_NK4A136_group, Clostridia_UCG-014_unclassified, Monoglobus, Phascolarctobacterium, Candidatus_Saccharimonas, Desulfovibrionaceae unclassified, and Desulfovibrio, and decreasing the abundance of Muribaculaceae_unclassified and Enterorhabdus. Lycium barbarum polysaccharide also significantly increased short-chain fatty acid levels, and Clostridia_UCG-014_unclassified, Candidatus_Saccharimonas, and Muribaculaceae_unclassified may participate in regulating butyric acid production to improve glycolipid metabolism in type 2 diabetic rats. Lachnospiraceae_NK4A136_group, Monoglobus, and Desulfovibrionaceae unclassified may participate in regulating isobutyric acid production to inhibit insulin resistance and improve lipid metabolism. Lycium barbarum polysaccharide can improve the inflammatory response in type 2 diabetic rats by increasing the abundance of Firmicutes_unclassified, Clostridia_UCG-014_unclassified, Intestinimona, and Colidextribacter, and inhibiting the abundance of Kineothrix.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21543
BACKGROUND: Animal experiments have demonstrated that Bushen Qiangjin capsule can significantly improve subchondral bone metabolism and abnormal bone remodeling, and delay the progression of knee osteoarthritis, providing experimental evidence for elucidating the mechanism of action of this drug in treating knee osteoarthritis. OBJECTIVE: To systematically explore the mechanism of action of Bushen Qiangjin capsule in regulating inflammatory signaling pathways to improve knee osteoarthritis using network pharmacology, Mendelian randomization based on pooled data, two-sample Mendelian randomization, molecular docking, and cell experiments. METHODS: (1) Network pharmacology tools were used to obtain the active components, core targets, and signaling pathways of Bushen Qiangjin capsule in treating knee osteoarthritis. Potential targets were imported into the STRING platform to construct a protein-protein interaction network and further screen core targets. Mendelian randomization based on pooled data was conducted to evaluate the causal relationship between core targets and the risk of knee osteoarthritis. Two-sample Mendelian randomization was used to analyze the causal relationship between mitogen-activated protein kinase 8 (MAPK8) and knee osteoarthritis. GO function and KEGG pathway enrichment analyses were performed on potential targets of Bushen Qiangjin capsule intervention in knee osteoarthritis. Molecular docking was used to further clarify the interaction between key active components of Bushen Qiangjin capsule and core target proteins. (2) Five SD rats were given Bushen Qiangjin capsule by gavage at 0.243 g/(kg·d) for 7 consecutive days. After the last administration, venous blood was collected and serum was separated to obtain drug-containing serum. Rat knee chondrocytes were divided into three groups for culture: blank group (no treatment), model group (treated with interleukin-1β for 24 h to establish an osteoarthritis cell model), and drug-containing serum group (treated with interleukin-1β for 24 h, then treated with 10% drug-containing serum for 24 h). After treatment, Alcian blue and toluidine blue staining were used to observe chondrocyte morphology. RT-qPCR and western blot were used to detect the mRNA and protein expression of MAPK8 and type II collagen. RESULTS AND CONCLUSION: (1) A total of 85 active components related to Bushen Qiangjin capsule were obtained, among which kaempferol and ferulic acid were key components. Core targets included serine/threonine-protein kinase 1 (AKT1), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), etc. Mendelian randomization based on pooled data and two-sample Mendelian randomization indicated that MAPK8 had a direct causal relationship with knee osteoarthritis and was positively correlated. GO enrichment results mainly involved biological processes such as phosphorylation, response to xenobiotic stimulus, and negative regulation of apoptotic process. KEGG pathways mainly included phosphatidylinositol 3-kinase-serine/threonine-protein kinase, tumor necrosis factor alpha, Toll-like receptor 4, hypoxia-inducible factor 1, and mechanosensory-related axes. Molecular docking results showed that kaempferol and ferulic acid had good binding activity with core targets such as AKT1, PIK3CA, tyrosine kinase C, signal transducer and activator of transcription 3, and tumor protein p53. (2) Alcian blue and toluidine blue staining showed that Bushen Qiangjin capsule could promote the synthesis of cartilage extracellular matrix in the osteoarthritis cell model. RT-qPCR and western blot showed that compared with the model group, the mRNA and protein expression of type II collagen increased (P < 0.05), while the mRNA and protein expression of MAPK8 decreased (P < 0.05) in the drug-containing serum group. (3) These results indicate that Bushen Qiangjin capsule may improve knee osteoarthritis-related damage through 'multi-component, multi-target, multi-pathway' synergistic regulation of inflammatory response and extracellular matrix homeostasis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21537
BACKGROUND: Recent studies have shown that exercise modulates gut microbiota and glucose metabolism; however, the mechanism linking exercise to gut microbiota and short-chain fatty acid production in type 2 diabetes remains unclear. OBJECTIVE: To investigate the mechanism by which exercise modulates gut microbiota composition and short-chain fatty acid metabolism to treat type 2 diabetes. METHODS: Twenty male Sprague-Dawley rats were randomly divided into a control group (n=6) and a model group (n=14). The rats in the model group were fed a high-sugar, high-fat diet for 8 weeks to induce insulin resistance. Following 12 hours of fasting (water allowed), rats received a tail vein injection of 1% streptozotocin solution (35 mg/kg) to damage pancreatic β-cells and elevate blood glucose, establishing type 2 diabetes models. Following successful modeling, feeding protocols remained unchanged. Twelve type 2 diabetes rats were divided into a model group (n=6) and an exercise group (n=6), and the exercise group were subjected to 12 weeks of aerobic exercise. Following the final aerobic exercise intervention, blood samples were collected for glucose metabolism indicators, and fresh feces were collected for short-chain fatty acid measurement by gas chromatography. Total microbial DNA was extracted from fresh feces, PCR amplified, purified, and sequenced using NovaSeq high-throughput sequencing. Species annotation was performed using the SILVA database, differential flora screening based on linear discriminant analysis effect size, and MetaCyc functional pathway prediction to explore associations between exercise intervention and glycolipid metabolism pathways. Heatmaps and visualization network diagrams were used to analyze correlations between key flora and biochemical indicators. RESULTS AND CONCLUSION: After 12 weeks of aerobic exercise, the exercise group showed significant improvement in glycolipid metabolism disorders, reduced inflammation, enhanced insulin sensitivity, and significantly decreased fasting blood glucose (P < 0.01). α-diversity analysis showed that the exercise group had significantly higher richness (Chao index), coverage (Coverage index), diversity (Shannon index), and evenness (Simpson index) of gut microbiota compared to the model group (P < 0.05). Abundance statistics, correlation heatmaps, and network diagrams showed that exercise significantly increased the abundance of short-chain fatty acid-producing genera such as Colidextribacter and Intestinimonas within Firmicutes, positively correlated with hexanoic acid and valeric acid levels; while inhibiting pathogenic bacteria such as Klebsiella and Turicibacter within Proteobacteria, negatively correlated with short-chain fatty acid levels. Lactobacillus promoted the production of butyric acid and other short-chain fatty acids, and was negatively correlated with glycolipid metabolism and inflammatory markers, fasting blood glucose, and interleukin-6 (P < 0.05). KEGG and MetaCyc metabolic function prediction showed that aerobic exercise reshaped energy homeostasis by bidirectionally regulating microbial metabolic pathways: significantly downregulating pathways related to excessive glycolipid catabolism and pro-inflammatory metabolism, while upregulating key pathways for short-chain fatty acid synthesis and glycolytic homeostasis. This functional remodeling was highly synergistic with the restoration of short-chain fatty acid-producing genera in Firmicutes and inhibition of pathogenic bacteria in Proteobacteria (P < 0.05). These results suggest that the dynamic balance of microbial metabolic pathways and host glycolipid metabolism improvement and inflammation alleviation form a closed-loop regulation, indicating that microbiota, short-chain fatty acid synthesis, and metabolic function remodeling are core mechanisms by which exercise improves the pathological process of diabetes.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21585
BACKGROUND: Previous studies have demonstrated neuroprotective potential of microRNA-23a-3p in traumatic brain injury. However, direct evidence is still lacking regarding whether this protective effect stems from its precise regulation of the M1/M2 polarization balance of microglia. OBJECTIVE: To clarify the expression changes of microRNA-23a-3p in mouse brain tissue after traumatic brain injury and to explore the specific mechanism by which it affects neurological function through regulating microglial polarization. METHODS: Eighty C57BL/6J mice were randomly assigned to four groups: a sham operation group, a traumatic brain injury group, a traumatic brain injury + agomir-NC group, and a traumatic brain injury + agomir-MicroRNA-23a-3p group. The traumatic brain injury model was established using the cortical impact method. The sham group did not receive cortical impact. The intervention groups received intracerebroventricular injection of agomir-NC or agomir-MicroRNA-23a-3p after modeling. Six mice from the sham and traumatic brain injury groups were analyzed at 1, 3, 7, and 14 days post-injury, and six mice from the other two groups were analyzed at 14 days post-injury. Neurological deficits were assessed using the modified neurological severity score (mNSS). Hematoxylin-eosin staining and Nissl staining were used to observe pathological changes in brain tissue and neurons. qRT-PCR and western blot were used to detect the expression levels of MicroRNA-23a-3p, M1 markers (CD16, CD86), M2 markers (CD206, arginase-1), and inflammatory cytokines (tumor necrosis factor-α and interleukin-10). Immunohistochemistry was used to evaluate microglial M1/M2 polarization and the aggregation of F4/80-positive cells in the injured area. RESULTS AND CONCLUSION: Compared with the sham group, the expression of MicroRNA-23a-3p in the traumatic brain injury group showed a "V"-shaped curve, with downregulation in the early phase and upregulation starting at 7 days post-injury. Upregulation of MicroRNA-23a-3p reduced the mNSS score in traumatic brain injury mice. Morphological results showed that upregulation of MicroRNA-23a-3p alleviated brain edema and neuronal damage. Molecular biology results showed that upregulation of MicroRNA-23a-3p promoted microglial polarization from M1 to M2 phenotype. These findings indicate that MicroRNA-23a-3p can promote neurological function recovery after traumatic brain injury in mice by regulating microglial polarization.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21565
BACKGROUND: The vicious cycle of osteoarthritis initiation and progression is driven by the combined effects of mechanical microenvironment disruption and collapse of osmotic pressure homeostasis. Sustained abnormal osmotic pressure disrupts chondrocyte homeostasis and markedly impairs the ability of bone marrow mesenchymal stem cells to differentiate into chondrocytes. Consequently, this compromises the regenerative capacity of cartilage and accelerates the degeneration of articular cartilage. OBJECTIVE: To develop a pathological osmotic pressure model for use in osmotic intervention experiments, in order to investigate the effects of osmotic pressure on chondrogenic differentiation of bone marrow mesenchymal stem cells and chondrocyte matrix metabolism, and to explore the role of imbalanced osmotic pressure within the joint cavity in the pathogenesis of osteoarthritis. METHODS: Bone marrow mesenchymal stem cells were isolated from 6- to 8-week-old rats and cultured to the third passage. Third-passage rat chondrocytes were revived and performed expansion culture. Physiological or pathological osmotic pressure regulating solutions were prepared by adding NaCl to the culture medium, and their biocompatibility was assessed via cell counting kit-8 assays. Bone marrow mesenchymal stem cells were treated with different osmotic pressure regulating solutions in chondrogenic induction medium for 7 or 14 days. Safranin O staining was used to identify the secretion of glycosaminoglycan (a cartilage marker). Quantitative real-time PCR was used to detect the expression of cartilage synthesis-related genes. The effects of physiological or pathological osmotic pressure regulating solutions on chondrocyte anabolic and catabolic metabolism after interleukin-1β inflammation induction were detected. Furthermore, RNA-seq was used to identify differentially expressed genes between the physiological and pathological osmotic pressure groups and perform enrichment analysis. RESULTS AND CONCLUSION: (1) Third-passage bone marrow mesenchymal stem cells were successfully isolated and cultured, and third-passage chondrocytes were successfully revived and expanded. (2) CCK-8 assay results showed that both pathological and physiological osmotic pressure regulating solutions had good biocompatibility. (3) Safranin O staining indicated that at days 7 and 14, the chondrogenic capacity of the physiological osmotic pressure group was significantly enhanced compared with the pathological osmotic pressure group. (4) qRT-PCR results further showed that compared with the interleukin-1β group and the pathological osmotic pressure group, the physiological osmotic pressure group significantly upregulated the expression of cartilage synthesis-related genes aggrecan and collagen type II alpha 1, while downregulating the expression of catabolism-related genes matrix metalloproteinase 13 and matrix metalloproteinase 3. (5) RNA-seq results showed that under physiological osmotic pressure conditions, multiple molecules and signaling pathways related to osteoarthritis pathogenesis were significantly downregulated.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05049-7
Dental-derived stromal cells (DSCs), including periodontal ligament stem cells (PDLSCs), dental pulp stem cells (DPSCs), stem cells from the apical papilla (SCAP), and stem cells from human exfoliated deciduous teeth (SHED), are accessible and expandable candidates for oral and craniofacial regeneration. Their therapeutic performance remains inconsistent because conventionally expanded cells are poorly adapted to in vivo mechanical cues. This review presents mechanical preconditioning as a mechanomedicine-guided strategy for ex vivo functional priming of DSCs. It summarizes DSC responses to tensile and compressive forces, fluid shear stress, hydrostatic pressure, matrix stiffness, and surface topography, and discusses principal mechanotransduction pathways. Representative quantitative loading windows are outlined to support subtype-specific and indication-specific preconditioning design. Key translational barriers include stromal cell heterogeneity, donor variability, senescence, uncertain persistence of mechanically induced states, safety concerns, and the lack of standardized manufacturing workflows. Clinical translation will require a shift from descriptive mechanobiology toward parameter-defined, indication-specific, and good manufacturing practice-compatible preconditioning strategies for DSC-based regeneration.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025221
Hypertension is commonly accompanied by endothelial dysfunction, characterized by an imbalance between vasodilatation and constriction, increased levels of the proinflammatory factors interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1), and decreased nitric oxide (NO) bioavailability. Using an angiotensin II (Ang II)-induced endothelial dysfunction model, we show that treatment with the hydrogen sulfide (H₂S) donor GYY4137 significantly reverses Ang II-induced damage. GYY4137 restores sirtuin 6 (SIRT6) expression, suppresses inflammation, and improves vasodilatory function. Furthermore, endothelial-specific cystathionine-γ-lyase (CSE)-deficient mice exhibit inflammation and endothelial dysfunction in blood vessels, which is reversed by H₂S supplementation. Critically, SIRT6 inhibitors block the protective effects of H₂S in the endothelium. This study demonstrates that H₂S protects vascular endothelial function by activating the SIRT6 anti-inflammatory pathway.