Key Takeaways & Executive Findings
- •• Asiaticoside improves bone microarchitecture and histology in ovariectomized osteoporotic rats. • Asiaticoside reduces serum inflammatory cytokines (IL-1β, IL-18) and downregulates NLRP3 inflammasome components. • Asiaticoside promotes osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro. • The mechanism involves upregulation of TRIM24 and inhibition of NLRP3 expression.
Abstract
BACKGROUND: Studies have indicated that asiaticoside possesses multiple pharmacological functions, including anti-inflammatory and antioxidant properties, and exerts a positive effect on osteogenic differentiation. It may serve as a potential therapeutic agent for osteoporosis. OBJECTIVE: To investigate the effects of asiaticoside on osteogenic differentiation in osteoporotic rats. METHODS: (1) Animal models of osteoporosis were established by removing both ovaries in 52 female Sprague-Dawley rats. Simultaneously, equivalent volumes of adipose tissue near the ovaries were removed bilaterally from 10 female Sprague-Dawley rats as the sham operation group. At 8 weeks after modeling, 50 model rats were randomly divided into five intervention groups: model group (n=10) was administered saline via oral gavage; low-dose asiaticoside group (n=10) was administered 16 mg/(kg·d) asiaticoside via oral gavage; high-dose asiaticoside group (n=10) was administered 32 mg/(kg·d) asiaticoside via oral gavage; positive control group (n=10) was administered alendronate sodium tablets 7.35 mg/(kg·d) via oral gavage; high-dose asiaticoside plus activator group (n=10) was administered 32 mg/(kg·d) asiaticoside plus 300 mg/(kg·d) NLRP3 activator via oral gavage, once daily for 6 weeks. After administration, serum levels of interleukin-1β and interleukin-18 were measured, Micro-CT scanning of the distal femur was performed, hematoxylin-eosin staining was used to observe histological changes, and western blot was used to detect the protein expression of tripartite motif-containing protein 24, NLRP3, and cleaved caspase-1 in the femur. (2) Rat bone marrow mesenchymal stem cells in logarithmic growth phase were divided into four groups: blank group (no treatment), asiaticoside group (treated with 20 µmol/L asiaticoside for 48 h), asiaticoside + empty vector group (transfected with empty vector plasmid for 48 h then treated with 20 µmol/L asiaticoside for 48 h), and asiaticoside + NLRP3 overexpression group (transfected with NLRP3 overexpression plasmid for 48 h then treated with 20 µmol/L asiaticoside for 48 h). After 7 days of osteogenic induction, alkaline phosphatase activity and mRNA expression of osteopontin, osteocalcin, tripartite motif-containing protein 24, and NLRP3 were detected. RESULTS AND CONCLUSION: (1) Animal experiments: The levels of interleukin-1β and interleukin-18 in the low-dose asiaticoside group, high-dose asiaticoside group, and positive control group were lower than those in the model group (P < 0.05), while the levels in the high-dose asiaticoside plus activator group were higher than those in the high-dose asiaticoside group (P < 0.05). Micro-CT scanning and hematoxylin-eosin staining results showed that compared with the model group, the bone microarchitecture and histological morphology of the femur were significantly improved in the low-dose asiaticoside group, high-dose asiaticoside group, and positive control group, while the NLRP3 activator partially inhibited the effects of high-dose asiaticoside. Compared with the model group, the protein expression of tripartite motif-containing protein 24 was increased (P < 0.05), and the protein expression of NLRP3 and cleaved caspase-1 was decreased (P < 0.05) in the low-dose asiaticoside group, high-dose asiaticoside group, and positive control group; the protein expression of tripartite motif-containing protein 24 in the high-dose asiaticoside plus activator group was lower than that in the high-dose asiaticoside group (P < 0.05), while NLRP3 and cleaved caspase-1 proteins were higher (P < 0.05). (2) Cell experiments: The alkaline phosphatase activity in the asiaticoside group was higher than that in the blank group and asiaticoside + NLRP3 overexpression group (P < 0.05). RT-PCR detection showed that the mRNA expression of osteopontin, osteocalcin, and tripartite motif-containing protein 24 in the asiaticoside group was higher than that in the blank group and asiaticoside + NLRP3 overexpression group (P < 0.05), while NLRP3 mRNA expression was lower (P < 0.05). (3) The results indicate that asiaticoside may promote osteogenic differentiation in osteoporotic rats by upregulating the expression of tripartite motif-containing protein 24 and inhibiting the expression of NLRP3, thereby delaying the progression of osteoporosis.
1. Introduction
Osteoporosis is a degenerative skeletal disease that increases bone fragility and susceptibility to fractures. According to the World Health Organization, osteoporosis ranks seventh among common diseases in the elderly, especially affecting postmenopausal women, seriously impacting patients' quality of life and imposing a heavy burden on healthcare [1-3]. Osteoporosis is caused by an imbalance between bone formation and bone resorption. Osteoblasts play a key role in maintaining the physiological functions of the human skeletal system and are the main functional cells for bone formation and metabolism. Therefore, focusing on osteoblast differentiation is beneficial for the treatment of osteoporosis [4].
Asiaticoside is a pentacyclic triterpene molecule extracted from Centella asiatica, a traditional Chinese medicine with broad pharmacological activities. It has high safety and few adverse reactions, and possesses anti-inflammatory, antioxidant, and other pharmacological functions, exerting a positive effect on osteogenic differentiation [5-8]. Studies have shown that inflammatory responses can regulate bone formation and resorption, playing a key role in the bone remodeling process of osteoporosis. The NOD-like receptor protein 3 (NLRP3) inflammasome is associated with the development of autoinflammatory diseases, and inhibiting the NLRP3 inflammasome can reduce the occurrence of postmenopausal osteoporosis [9-10]. Tripartite motif-containing protein 24 (TRIM24) is a negative regulator of inflammatory responses and can ubiquitinate and degrade NLRP3 [11]. This study aims to investigate the effects of asiaticoside on osteogenic differentiation in osteoporotic rats and its underlying mechanism.
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Xu Hongtao, Wang Jianping, Xu Yuehong, Li Qin, Qi Qihua, Xia Qipeng (2026). Asiaticoside promotes osteogenic differentiation in osteoporotic rats. Chinese Journal of Tissue Engineering Research. https://doi.org/10.12307/2026.21560
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Frequently Asked Questions
What is the main finding of this study?
The study demonstrates that asiaticoside promotes osteogenic differentiation in osteoporotic rats by upregulating TRIM24 expression and inhibiting NLRP3 expression, thereby delaying osteoporosis progression.
How was the osteoporosis model established in this study?
Osteoporosis was induced in female Sprague-Dawley rats by bilateral ovariectomy, and a sham operation group was created by removing equivalent volumes of adipose tissue near the ovaries.
What doses of asiaticoside were used in the animal experiments?
Low-dose asiaticoside group received 16 mg/(kg·d) and high-dose group received 32 mg/(kg·d) via oral gavage for 6 weeks.
What are the key molecular markers assessed in this study?
The study assessed serum levels of IL-1β and IL-18, protein expression of TRIM24, NLRP3, and cleaved caspase-1, as well as mRNA expression of osteopontin, osteocalcin, TRIM24, and NLRP3.
What is the potential mechanism of asiaticoside in treating osteoporosis?
Asiaticoside likely promotes osteogenic differentiation by upregulating TRIM24, which in turn inhibits NLRP3 inflammasome activation, reducing inflammation and enhancing bone formation.
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