伸筋草对类风湿性关节炎大鼠踝关节滑膜组织ICAM-1、MMP-3及OPG/RANKL/RANK信号通路的影响

Effect of Lycopodium clavatum on ICAM-1,MMP-3 and OPG/RANKL/RANK Signal Pathway in Synovium Tissue of Ankle Joint in Rats with Rheumatoid Arthritis

目的:探讨伸筋草对类风湿性关节炎大鼠踝关节滑膜组织细胞间黏附分子-1、基质金属蛋白酶-3及OPG/RANKL/RANK信号通路的影响。方法:弗氏完全佐剂皮下注射复制大鼠类风湿性关节炎模型。根据模型复制情况将Wistar雄性大鼠按照体质量区组化随机分为6组,即模型组、伸筋草高、中、低给药组、阳性对照组和正常对照组,每组10只。各给药组每日灌胃给药相应剂量的伸筋草提取物浓缩液(1.6 g/kg、0.8 g/kg、0.4 g/kg),阳性对照组给予雷公藤多苷溶液(5.4 mg/kg),正常对照组、模型组大鼠灌胃等体积的生理盐水,连续3周。光镜下观察大鼠踝关节滑膜组织病理变化,采用免疫组化法测定各组大鼠滑膜组织ICAM-1和MMP-3的表达,Western Blot法测定滑膜组织中OPG、RANKL蛋白的表达,并推算RANKL/OPG的比值。结果:与空白对照组相比,模型组大鼠滑膜组织有明显的淋巴细胞及单核细胞等炎性浸润,滑膜组织细胞排列不规则,增生肿胀明显,ICAM-1、MMP-3水平和OPG、RANKL蛋白表达以及RANKL/OPG比值明显增加(P<0.05);与模型相比,伸筋草高、中、低剂量组大鼠滑膜组织增生和炎性细胞浸润均有改善的趋势,ICAM-1、MMP-3水平、OPG、RANKL蛋白表达以及RANKL/OPG比值也随剂量的增加而降低。高剂量组改善效果最为明显,中剂量组次之,低剂量组虽有改善的趋势,但无统计学差异(P>0.05)。结论:伸筋草可以降低大鼠踝关节滑膜组织炎性因子的水平,并通过调控OPG/RANKL/RANK信号通路来发挥治疗RA的作用。

Objective:To investigate the effect of Lycopodium clavatum on the expressions of ICAM-1,MMP-3 and OPG/RANKL/RANK signal pathway in synovium tissue of ankle joint in rats with rheumatoid arthritis(RA).Methods:RA model was duplicated by the injection of Freund’s complete adjuvant subcutaneously.After RA model was established,Wistar male rats were randomly divided into 6 groups according to body weight,i,e.the model group,the Lycopodium clavatum groups of high-dose,medium-dose and low-dose,the positive control group and the normal control group,with 10 rats in each group.Rats in administrated group received corresponding dose of Lycopodium clavatum(1.6 g/kg,0.8 g/kg,0.4 g/kg)and Tripterygium glycoside(5.4 mg/kg)once a day for 3 weeks.Rats in the normal group and the model group received the same volume of saline during treatment period.Histopathological changes of synovium in ankle joint were observed under light microscope.The expressions of ICAM-1 and MMP-3 in synovium of rats were evaluated via immunohistochemistry in each group.Western blot was used to determine the expressions of OPG and RANKL in synovium.The ratio of RANKL/OPG was calculated as well.Results:Compared to the normal control group,the synovium tissue in the model group had obvious inflammatory infiltration of lymphocytes and monocytes,irregular arrangement and marked proliferation and swelling of synovium tissue cells could be seen.The levels of ICAM-1 and MMP-3,as well as the protein expressions of OPG and RANKL and ratio of RANKL/OPG increased significantly(P<0.05).Compared to the model group,the proliferation and inflammatory cell infiltration of synovium in the groups of high-dose,medium-dose and low-dose were ameliorated.The levels of ICAM-1 and MMP-3,the expressions of OPG,RANKL and the ratio of RANKL/OPG also decreased with the increasing dose of Lycopodium clavatum.High-dose group showed the best efficacy,followed by the medium-dose group.Although there was a trend performed by low-dose group,no statistical difference showed by ANVOA(P>0.05).Conclusions:Lycopodium clavatum is effective in the treatment of RA.The mechanism is related to reducing the level of inflammatory factors and regulating OPG/RANKL/RANK signal pathway in synovium tissue.