牛膝总皂苷对实验兔膝骨关节炎滑膜组织的影响

Effect of Total Saponins of Achyranthes on Osteoarthritis Synovial Tissue in Experimental Rabbits

目的:观察牛膝总皂苷(TSA)对兔膝骨关节炎(OA)模型滑膜组织的影响。方法:将40只实验兔分为正常组8只、OA模型组32只,OA造模成功后将OA模型组分为OA模型阳性对照组、OA模型空白组、OA模型TSA低剂量组、OA模型TSA中剂量组、OA模型TSA高剂量组,各6只,分别予硫酸氨基葡萄糖、蒸馏水、TSA低剂量、TSA中剂量、TSA高剂量灌胃2个月后进行关节粘连改善度测定、滑膜组织病理HE染色、滑膜炎KRENN评分、β-catenin蛋白免疫组化检测,及关节液TNF-1,IL-1β,MMP-3含量检测。结果:实验兔OA模型造模成功后灌胃2个月,与OA模型空白组关节粘改善度比较,OA模型阳性对照组和OA模型TSA低、中、高剂量组差异有统计学意义(P<0.05),OA模型阳性对照组与OA模型TSA高剂量组比较差异有统计学意义(P<0.01);滑膜HE染色和滑膜炎评分比较,OA模型空白组见大量滑膜组织炎细胞浸润、纤维组织增生以及滑膜细胞增生、排列紊乱,滑膜组织周边可见较多毛细血管增生,呈重度滑膜炎表现,OA模型阳性对照组见中度滑膜组织炎细胞浸润、纤维组织增生以及滑膜细胞增生、排列不规则,滑膜组织周边可见少量毛细血管增生,呈中度滑膜炎表现,OA模型TSA高剂量组见轻度滑膜组织炎细胞浸润、纤维组织增生以及滑膜细胞增生、排列尚规则,滑膜组织周边可见微量毛细血管增生,呈轻度滑膜炎表现;β-catenin蛋白免疫组化比较,OA模型阳性对照组和OA模型TSA高剂量组与OA模型空白组比较,差异有统计学意义(P<0.01),OA模型阳性对照组与OA模型TSA高剂量组比较差异有统计学意义(P<0.05);各组与正常组比较TNF-1,IL-1β,MMP-3指标差异有统计学意义(P<0.05),OA模型TSA高剂量组与OA模型阳性对照组比较差异无统计学意义(P>0.05),两者与OA模型空白组比较差异有统计学意义(P<0.01)。结论:牛膝总皂苷能有效抑制OA滑膜炎症,改善关节粘连度,降低滑膜组织β-catenin蛋白表达以及减少关节液TNF-1,IL-1β,MMP-3因子含量,其作用机理有待进一步探索。

Objective： To observe the effect of total saponins of achyranthes（TSA）on synovial tissue in osteoarthritis（OA）model of rabbit. Methods： Forty experimental rabbits were divided into normal group（n=8）and OA model group（n=32）. The OA model group was divided into OA model positive control group, OA model blank group, and OA model TSA low dose group, medium dose group, and high dose group（6 cases in each group）after successful modeling in OA, which were treated with glucosamine sulfate, distilled water and low dose, medium dose and high dose of TSA by gavage respectively. After 2 months, the improvement degree of joint adhesion was measured. The synovial tissue was staining by pathology of hematoxylin-eosin（HE）. The KRENN score of synovitis was recorded. The expression of β-catenin was detected by immunohistochemistry method. The levels of TNF-1, IL-1β and MMP-3 in synovial fluid were detected by Elisa method. Results： After successful modeling in OA and intragastrical administration for 2 months, the improvement degree of joint adhesion in the OA model positive control group and the OA model TSA low, medium and high dose group were statistical different than the OA model blank group（P〈0.05）, and there was significantly different between the OA model positive control group and the OA model TSA high dose group（P〈0.01）. Comparison of synovial HE staining and synovitis score： OA model blank grup showed a large number of synovial tissue inflammatory cell infiltration, fibrous tissue hyperplasia and synovial cell hyperplasia, derangement, synovial tissue surrounding more visible capillary hyperplasia, which showed severe manifestations of synovitis. In OA model positive control group, there was moderate synovial tissue inflammatory cell infiltration, fibrous tissue hyperplasia and synovial cell hyperplasia, irregular arrangement, synovial tissue surrounding a small amount of blood capillary hyperplasia, synovitis showed moderate expression. OA model TSA high dose group showed mild synovial inflammatory cell infiltration, fibrous tissue hyperplasia and synovial cell hyperplasia, row column still rules, synovial tissue could be seen around the micro capillary hyperplasia, mild manifestations of synovitis. Compared with OA model blank group on β-catenin protein immunohistochemistry, OA positive control group and OA model TSA high dose group had significantly statistical different（P〈0.01）, and there was significant difference between OA model positive control group and OA model TSA high dose group（P〈0.05）. Compared with the normal group on TNF-1, IL-1β, MMP-3 index, the other groups had significant different（P〈0.05）, and the OA model positive control group and OA model TSA high dose group had no significant difference（P〉0.05）, both of them were significant difference when compared with OA model blank group（P〈0.01）. Conclusion： Total saponins of achyranthes can effectively inhibit OA synovial inflammation, improve the degree of articular adhesion, reduce the expression of β-catenin protein in synovial tissue and decrease the levels of IL-1β, TNF-1, MMP-3 factor in joint fluid, and its mechanism should be further explored.