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目的观察骨关节炎(OA)成纤维样滑膜细胞(FLS)体外培养时生长增殖特性,以及白芍总苷(TGP)对其增殖能力的影响,探讨白芍总苷的抗炎机制。方法四甲基偶氮唑蓝(MTF)法和反转录-聚合酶链反应(RT-PCR)方法观察OA和正常对照(NS)膝关节滑膜组织体外培养时FLS增殖能力和c-fos表达情况,酶联免疫吸附试验(ELISA)法检测培养上清中的肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ和碱性成纤维细胞生长因子(bFGF),用流式细胞仪测定FLS细胞周期的变化。结果OA和NS的FLS在体外培养时的细胞倍增时间差异无统计学意义。加入高剂量TGP对OA-FLS细胞存活分数(SF)的抑制作用较NS-FLS组更显著(P<0.05),2000 mg/L和400 mg/L TGP降低培养液上清中TNF-α和bFGF的水平,升高IFN-γ含量(P<0.05),并抑制FLS的c-los mRNA表达(P<0.05)。OA-FLS细胞周期测定表明加入2000 mg/L的TGP可延长G_1期而缩短S期,但与不加药对照组相比差异无统计学意义(P>0.05)。结论高浓度TGP对OA-FLS的增殖能力、细胞因子分泌及原癌基因表达的抑制程度大于对正常滑膜FLS的作用,表明TGP调节FLS增殖特性,有抑制OA滑膜炎的作用。
Objective To observe the growth and proliferation characteristics of osteosarcoma (OA) fibroblast-like synoviocytes (FLS) cultured in vitro, and the effects of total glucosides of paeony (TGP) on its proliferative capacity, and to explore the anti-inflammatory mechanism of total glucosides of paeony. Methods MTT assay and reverse transcription-polymerase chain reaction (RT-PCR) were used to observe the proliferation of FLS and c-fos in OA and normal control (NS) knee synovial tissues in vitro. Expression, TNF-α, interferon (IFN)-γ, and basic fibroblast growth factor (bFGF) in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA) using flow cytometry The instrument measures changes in FLS cell cycle. Results There was no significant difference in doubling time of OA and NS FLS cultured in vitro. The addition of high-dose TGP inhibited the survival fraction (SF) of OA-FLS cells more significantly than that of NS-FLS group (P<0.05). 2000 mg/L and 400 mg/L TGP decreased the TNF-α in the culture supernatant. The level of bFGF increased IFN-γ content (P<0.05) and inhibited the expression of c-los mRNA in FLS (P<0.05). The OA-FLS cell cycle assay showed that the addition of 2000 mg/L TGP could prolong the G 1 phase and shorten the S phase, but the difference was not statistically significant (P>0.05). Conclusion The high concentration of TGP inhibited the proliferation of OA-FLS, the secretion of cytokines and the expression of proto-oncogenes more than the normal synovial FLS, indicating that TGP regulates the proliferation of FLS and inhibits the synthesis of OA synovitis.