刺山柑对类风湿关节炎大鼠滑膜组织自噬相关蛋白表达的影响

Effect of Capparis spinosa on the Expression of Autophagy-related Proteins in Synovial Tissues of Rats with Rheumatoid Arthritis

目的:观察刺山柑果风湿止痛贴对类风湿关节炎大鼠滑膜组织自噬相关蛋白表达的影响,并探究刺山柑治疗类风湿关节炎(RA)的可能作用机制。方法:将40只无特定病原体(SPF)级SD大鼠采用随机数字法分为正常组,模型组,阳性药组和刺山柑组,每组10只,对造模的大鼠在足跖底部注射0.1 mL完全弗氏佐剂。模型组、阳性药组和刺山柑组分别于大鼠左后足跖部外敷空白凝胶、伤湿止痛膏以及刺山柑果风湿止痛贴,每组持续干预21 d。观测关节炎(AI)指数和足爪容积的指标,酶联免疫吸附测定(ELISA)法检测大鼠血清肿瘤坏死因子-α(TNF-α)水平,蛋白质印迹法检测滑膜组织中微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)、Beclin蛋白复合物-1(Beclin-1)蛋白表达。结果:与正常组比较,模型组和阳性药组LC3-Ⅱ、Beclin-1蛋白表达均上升(P<0.05,P<0.01),刺山柑组LC3-Ⅱ蛋白表达降低(P<0.01),Beclin-1蛋白表达升高(P<0.01);与模型组比较,阳性药组Beclin-1、LC3-Ⅱ蛋白表达差异无统计学意义,刺山柑组LC3-Ⅱ蛋白表达降低(P<0.01),Beclin-1蛋白表达升高(P<0.01)。结论:刺山柑果风湿止痛贴治疗AA大鼠比伤湿止痛膏疗效更加显著,从而为刺山柑治疗RA提供一定的科学依据,其相关机制可能与蛋白LC3-Ⅱ、Beclin-1表达有关。

Objective:To observe the effect of Capparis spinosa fruit paste on the expression of autophagy-related proteins in synovial tissues of rats with rheumatoid arthritis,and to explore the possible mechanism of C.spinose in RA.Methods:Forty SPF rats were randomly divided into a normal group,a model group,a positive drug group,and a C.spinose group,with 10 rats in each group.The rats to be modeled were injected with 0.1 mL complete Freund′s adjuvant at the foot bottom.The model group,positive drug group,and C.spinose group externally applied blank gel,Shangshi Zhitong Ointment,and C.spinosa fruit paste,respectively,on the left posterior plantar of rats for 21 d.Arthritis index(AI)and paw volume index were measured.The level of serum tumor necrosis factor-α(TNF-α)was detected by enzyme-linked immunosorbent assay(ELISA),and the protein expression of microtubule-associated protein light chain 3-Ⅱ(LC3-Ⅱ)and Beclin-1 in synovial tissues was detected by Western blot.Results:As compared with the normal group,the protein expression of LC3-Ⅱand Beclin-1 in the model group and positive drug group was increased(P<0.05,P<0.01),and that of LC3-Ⅱwas decreased(P<0.01)while Beclin-1 was increased(P<0.01)in the C.spinose group.As compared with the model group,the protein expression of Beclin-1 and LC3-Ⅱin the positive drug group had no significant difference,while that of LC3-Ⅱwas decreased(P<0.01)and Beclin-1 protein was increased(P<0.01)in the C.spinose group.Conclusion:C.spinosa fruit paste is more effective in the treatment of rats with RA than Shangshi Zhitong Ointment,thus providing a scientific basis for the treatment of RA by C.spinosa,and the related mechanism may be related to the expression of LC3-Ⅱand Beclin-1.