牡荆素对Ⅱ型胶原蛋白诱导的大鼠类风湿关节炎的缓解作用及对血浆Th17、Th23和MMPs蛋白水平的影响

Effect of vitexin on type Ⅱ collagen-induced rheumatoid arthritis in rovt and plasma Th17,Th23 and MMPs protein levels

目的:研究牡荆素对Ⅱ型胶原蛋白诱导的大鼠类风湿关节炎(RA)的作用及对血浆Th17、Th23和MMPs蛋白水平的影响。方法:大鼠尾根部分皮内注射牛Ⅱ型胶原乳液造模,造模52 d后,给药组每天分别腹腔注射牡荆素1.5、3、6 mg/kg,连续3周;HE染色观察软骨组织病理损伤,Western blot检测Ki67、PCNA、Caspase-3、Caspase-9、MMP-1、MMP-3、MMP-13、TGF-β1和Smad2蛋白表达水平,ELISA检测外周血IL-17和IL-23的表达,流式分选CD4^+IL-17^+和CD4^+IL-23^+,RT-PCR检测MMP-1、MMP-3、MMP-13表达水平。结果:与Model组相比,给药组病理症状明显减轻,Ki67和PCNA蛋白表达显著上调,Caspase-3和Caspase-9蛋白表达显著下调,IL-17和IL-23的含量显著降低,CD4^+IL-17^+和CD4^+IL-23^+百分比显著降低,MMP-1、MMP-3和MMP-13 RNA表达显著下调,MMP-1、MMP-3和MMP-13蛋白表达显著下调,TGF-β1和p-Smad2蛋白表达显著下调,且呈剂量依赖性增加。结论:牡荆素对Ⅱ型胶原蛋白诱导的大鼠RA具有缓解作用,且下调RA大鼠血浆中Th17、Th23和MMPs蛋白表达。

Objective:To investigate the effects of vitexin on type Ⅱ collagen-induced rheumatoid arthritis(RA) in rats and the effects of plasma levels of Th17,Th23 and MMPs.Methods: The rheumatoid arthritis model was established by intradermal injection of bovine type Ⅱ collagen emulsion through the tail of rats.Drug-adminitered group was intraperitoneally injected with vitexin 1.5,3,6 mg/kg daily after 52 days of modeling,continuous for 3 weeks.Pathological damage of cartilage was observed by HE staining,the expression levels of Ki67,PCNA,Caspase-3,Caspase-9,MMP-1,MMP-3,MMP-13,TGF-β1 and Smad2 were detected by Western blot,the expressions of IL-17 and IL-23 in peripheral blood were detected by ELISA.CD4^+IL-17^+ and CD4^+IL-23^+were sorted by flow cytometry,the expression levels of MMP-1,MMP-3 and MMP-13 were detected by RT-PCR.Results: Compared with model group,the pathological symptoms of drug-administered group were significantly reduced,the expression of Ki67 and PCNA protein was significantly increased,the expression of Caspase-3 and Caspase-9 protein was significantly decreased,and the levels of IL-17 and IL-23 were significantly decreased.The percentages of CD4^+IL-17^+and CD4^+IL-23^+were significantly decreased,MMP-1,MMP-3 and MMP-13 RNA expression were significantly decreased,MMP-1,MMP-3 and MMP-13 protein expression,TGF-β1 and p-Smad2 protein expression were significantly decreased,and these effects increased as the amount administered.Conclusion: Vitexin has a remission effect on type Ⅱ collagen-induced RA arthritis in rats,and down-regulates the expression of Th17,Th23 and MMPs in plasma of RA rats.