草乌甲素对大鼠胶原诱导性关节炎的作用

Effect of Bulleyaconitine A on collagen-induced arthritis in rats

目的 探究草乌甲素对大鼠胶原诱导性关节炎的作用。方法 20只雄性SPF级SD大鼠随机选取5只作为对照组,剩下的15只SD大鼠用来建立胶原诱导性关节炎(CIA)模型,根据关节炎评分表评估CIA模型,建模成功后再随机分成模型组、低剂量草乌甲素治疗组(50μg/kg)、高剂量草乌甲素治疗组(100μg/kg),此后每组大鼠连续给药干预30 d,第31天后处死大鼠。酶联免疫吸附法(ELISA)测定大鼠TNF-α、IL-6、IL-17、MMP-3水平,逆转录聚合酶链式反应(RT-PCR)测定关节滑膜组织中MMP-3 mRNA表达水平,蛋白印迹法(Western Blot)检测关节滑膜组织中MMP-3蛋白水平,HE染色观察滑膜组织病理变化。结果 与对照组比较,模型组及草乌甲素治疗组TNF-α、IL-6、IL-17、MMP-3水平明显升高(P<0.05);同时,与模型组比较,草乌甲素治疗组TNF-α、IL-6、IL-17、MMP-3水平明显降低(P<0.05)。与对照组相比,模型组MMP-3 mRNA、MMP-3蛋白表达水平明显升高;同时,与模型组比较,草乌甲素治疗组MMP-3 mRNA、MMP-3蛋白表达水平降低(P<0.05)。组织病理结果显示草乌甲素治疗组的关节滑膜处的炎症细胞浸润及滑膜增生情况均减轻。结论 草乌甲素可以抑制炎症因子分泌和降低MMP-3的表达,对大鼠胶原诱导性关节炎具有一定的治疗作用。

Objective To investigate the effect of Bulleyacon-itine A on collagen-induced arthritis rats.Methods Five of the 20 male SPF SD rats were randomly selected as the control group,and the remaining 15 SD rats were used to establish the collagen-induced arthritis(CIA)model.The CIA model was evaluated against the arthritis rating scale.After successful modeling,they were randomly divided into model group,low-dose aconitine treatment group(50μg/kg)and high-dose aconitine treatment group(100μg/kg).The rats in each group were given continuous intervention for 30 days,and the rats were killed on the 31st day.The levels of TNF-α,IL-6,IL-17 and MMP-3 were determined by enzyme-linked immunosorbent assay(ELISA),and the mRNA expression of MMP-3 in synovium was determined by reverse transcription polymerase chain reaction(RT-PCR).The level of MMP-3 protein in synovial tissue was detected by Western Blot,and the pathological changes of synovial tissue were observed by HE staining.Results Compared with control group,the levels of TNF-α,IL-6,IL-17 and MMP-3 in model group and aconitine treatment group were significantly increased(P<0.05).Compared with model group,the contents of TNF-α,IL-6,IL-17 and MMP-3 in monoponitine treatment group were significantly decreased(P<0.05).Histopathological results showed that the inflammatory cell infiltration and synovial hyperplasia were reduced in the joint synovium treated group.Conclusion Bulleyaconitine A can inhibit the secretion of inflammatory factors and reduce the expression of MMP-3,which has a therapeutic effect on collagen-induced arthritis in rats.