苗药金乌健骨方对类风湿关节炎滑膜血管新生的影响

Effects of Miao medicine Jinwu Jiangu Formula on synovial angiogenesis in rheumatoid arthritis

目的:探讨金乌健骨方总提物对类风湿关节炎成纤维样滑膜细胞(RA-FLS)滑膜血管新生因子的影响。方法:制备金乌健骨方总提物,将培养的RA滑膜细胞分为金乌健骨方低、中、高剂量组,来氟米特组,雷公藤多苷组和空白对照组,给药干预24h后,采用MTT法检测金乌健骨方对RA-FLS生长抑制率,ELISA法检测致炎因子IL-1β、IL-6、IL-8、TGF-β的分泌,PCR法检测血管生成相关基因VEGF、Ang、HIF-1α、HIF-1βmRNA的表达量。结果:给予不同浓度金乌健骨方总提物干预24h后,随着金乌健骨方浓度的升高,OD值逐渐降低,细胞生长抑制率逐渐增加。ELISA检测结果显示各给药组IL-1β、IL-6、IL-8、TGF-β分泌量均有下降,与空白对照组比较差异具有统计学意义(P<0.01),以金乌健骨方高剂量组最明显,与雷公藤多苷组和来氟米特组比较差异具有统计学意义(P<0.01)。PCR检测结果显示,给药干预后VEGF、Ang、HIF-1α、HIF-1βmRNA的表达水平降低,与空白对照组比较差异有统计学意义(P<0.01),金乌健骨方高剂量组下调VEGF、Ang、HIF-1α、HIF-1βmRNA作用优于雷公藤多苷组(P<0.01)。结论:金乌健骨方总提物能够抑制RA-FLS的增殖,下调致炎因子IL-1β、IL-6、IL-8、TGF-β的分泌,降低血管新生因子VEGF、Ang、HIF-1α、HIF-1β的表达。

Objective:To investigate the effect of Jinwu Jiangu Formula total extract on angiogenesis factor of human rheumatoid arthritis synovial fibroblasts(RA-FLS).Methods:The total extract of Jinwu Jiangu Formula was prepared.The cultured RA synovial cells were divided into Jinwu Jiangu Formula high,medium and low dose group,Tripterygium group,leflunomide group,and serum control group.After 24 hours of intervention,the inhibition rate of Jinwu Jiangu Formula on the growth of RA-FLS was detected by MTT method.The secretion of IL-1β,IL-6,IL-8 and TGF-βwas detected by ELISA.The expressions of VEGF,Ang,HIF-1 detected by mRNA were detected by PCR.Results:After 24 hours of intervention with different concentrations of total extracts of Jinwu Jiangu Formula,with the increase of Jinwu Jiangu Formula concentration,OD value decreased gradually and the cell growth inhibition rate also increased gradually.The results of ELISA showed that each group secretion of IL-1β,IL-6,IL-8,TGF-βwere reduced to varying degrees,its had significant difference compared with the control group(P<0.01).The high dose group of Jinwu Jiangu Formula was the most obvious,and had statistical difference compared with Tripterygium wilfordii polyglycoside group and leflunomide group(P<0.01).The results of PCR showed that the expression of VEGF,Ang,HIF-1α,HIF-1βmRNA were decreased after the intervention,which had statistical significance compared with the control group(P<0.01).Compared with Tripterygium wilfordii polyglycoside group,the expression of VEGF,Ang,HIF-1α,HIF-1βmRNA of the lyco high dose group of Jinwu Jiangu Formula were significantly reduced(P<0.01).Conclusion:The total extract of Jinwu Jiangu Formula can inhibit the proliferation of RA-FLS,decrease the secretion of pro-inflammatory cytokines by IL-1β,IL-6,IL-8,TGF-β,and reduce the expression of angiogenic factors VEGF,Ang,HIF-1α,HIF-1β.