人参皂苷Rg1对IL-1β诱导的人膝软骨细胞降解保护作用机制研究
The Protective Effect of Ginsenoside Rg1 on the Degradation of Knee Cartilage Induced by IL-1β and Its Related Mechanism
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摘要: 目的 研究人参皂苷Rg1对IL-1β诱导的人膝软骨降解的保护作用及其分子机制。方法 收集OA病人滑膜及软骨组织,培养成纤维滑膜细胞(FLS),采用IL-1β激发FLS细胞并与软骨骨片共培养,设空白对照组,IL-1β组(20 ng/mL)为阳性对照组,不同浓度Rg1(0、5、50 μmol/L)+IL-1β(20 ng/mL)为实验组。用番红O染色观察软骨降解情况,qPCR分析MMPs相关基因表达情况,ELISA法检测上清液中MMP-3表达情况,Western blot分析Rg1对OA-FLS细胞中NF-κB通路中相关蛋白的调控。结果 Rg1可以显著抑制IL-1β诱导的软骨降解,发挥软骨保护作用。Rg1对软骨保护作用主要通过抑制成纤维滑膜细胞中MMPs表达,其中主要抑制MMP-3基因和蛋白表达,且该作用可能与Rg1调控NF-κB通路有关。结论 人参皂苷Rg1可以抑制IL-1β诱导的软骨降解,发挥软骨保护作用,主要通过抑制FLS中MMP-3基因和蛋白表达。Abstract: OBJECTIVE To investigate the inhibition effects and underlying mechanism of Ginsenoside Rg1 on IL-1β induced cartilage degradation. METHODS The synovial membrane and cartilage tissue of Osteoarthritis patients were collected and cultured into fibroblast synoviocytes. The FLS cells were stimulated by IL-1β, and co-cultured with chondrocytes. We divided the experiment into different groups, including the blank group, the IL-1β group and the different concentration Rg1 groups. We observed the degradation of cartilage tissue was observed by staining with red O, and real-time fluorescence quantitative PCR was used to analyze the gene expression of MMPs, and the expression of MMP-3 in the supernatant was detected by ELISA, also the Western blot analysis was employed to determine the related proteins in NF-κB pathway. RESULTS Rg1 significantly inhibited the degradation of cartilage induced by IL-1β to protect the cartilage. The protective effect of Rg1 on cartilage was mainly contributed to its inhibition on MMPs, especially by inhibiting the MMP-3 gene and protein expression in the synovial fibroblasts. Rg1 inhibited the expression of MMP-3 mainly through NF-κB pathway. CONCLUSION Rg1 can inhibit the cartilage degradation induced by IL-1β and play the role of cartilage protection, mainly by inhibiting the expression of MMP-3 gene and protein in FLS.
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Key words:
- Ginsenoside Rg1 /
- IL-1β /
- cartilage protection /
- fibroblast synoviocytes
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