Authors
Chang-Long Fu, Dan-Ling Chen, Jing-Yi Guo, Shu-Jie Lan, Jia-Nan Chen, Xiao-Qian Ye
Published in
Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica. Volume 51. Issue 12. Pages 3532-3539.
Abstract
This study explored the mechanism by which Tougu Xiaotong Capsules(TGXTC) improve osteoarthritis(OA) degeneration, focusing on the regulation of cholesterol metabolism in OA chondrocytes via microRNA-16-5p(miR-16-5p). In vivo experiments were performed. A total of fifty 8-week-old C57BL/6 mice were acclimated for one week and then randomly divided into a blank group(10 mice) and a modeling group(40 mice). The OA model was established in the modeling group using the modified Hulth method. These mice were then randomly subdivided into four groups(n=10 per group): model, TGXTC, miR-16-5p antagomir, and TGXTC+miR-16-5p antagomir. Mice in the antagomir groups received intra-articular injections of miR-16-5p antagomir. After four weeks of intervention, cartilage structure was observed using Masson's trichrome, hematoxylin-eosin(HE), safranin O-fast green, and toluidine blue staining. The mRNA levels of miR-16-5p and cholesterol metabolism-related factors-ATP-binding cassette transporter A1(ABCA1), apolipoprotein A1(ApoA1), liver X receptor beta(LXRβ), sterol regulatory element-binding protein(SREBP), cholesterol-25-hydroxylase(CH25H), 25-hydroxycholesterol-7-alpha-hydroxylase(CYP7B1), and endoplasmic reticulum stress-associated factors C/EBP homologous protein(CHOP) and caspase-3, were detected by quantitative real-time PCR(RT-qPCR). The protein expression levels of ABCA1, ApoA1, LXRβ, SREBP, CH25H, CYP7B1, CHOP, and caspase-3 were analyzed by Western blot. In vitro experiments were also conducted. A chondrocyte degeneration model was established by stimulating mouse chondrocytes with thapsigargin(TG). RT-PCR was used to examine the levels of ABCA1, ApoA1, LXRβ, SREBP, CH25H, CYP7B1, CHOP and caspase-3 under miR-16-5p inhibition conditions. Flow cytometry was employed to assess the effect of TGXTC on the apoptosis rate of TG-induced chondrocytes following miR-16-5p inhibition. Histological staining results showed that, compared to the model group, the TGXTC group exhibited relatively clear and intact cartilage layer structure. Furthermore, compared to the miR-16-5p antagomir group, the TGXTC+miR-16-5p antagomir group showed improved cartilage surface damage. RT-PCR results indicated that, compared to the model group, the TGXTC group had significantly increased mRNA levels of miR-16-5p and ABCA1, ApoA1, and LXRβ, while the mRNA levels of SREBP, CH25H, CYP7B1, CHOP, and caspase-3 decreased. Western blot results showed that, compared to the model group, the TGXTC group showed upregulated protein expression of ABCA1, ApoA1, and LXRβ, and downregulated protein expression of SREBP, CH25H, CYP7B1, CHOP, and caspase-3. The RT-qPCR results for the in vitro experiments showed that in miR-16-5p-inhibited chondrocytes, TGXTC intervention increased the mRNA levels of ABCA1, ApoA1, and LXRβ, and decreased those of SREBP, CH25H, CYP7B1, CHOP, and caspase-3. Flow cytometry results confirmed that TGXTC mitigated the apoptosis of TG-induced chondrocytes. In conclusion, TGXTC can alleviate cholesterol metabolism disorders in osteoarthritic chondrocytes by regulating miR-16-5p.
PMID:
42543312
Bibliographic data and abstract were imported from PubMed on 03 Aug 2026.
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