摘要
目的 探讨贝母素乙调节Ras同源基因家族成员A(RhoA)/Rho相关的卷曲螺旋激酶(ROCK)信号通路对膝骨关节炎(knee osteoarthritis,KOA)大鼠软骨损伤的影响.方法 以碘乙酸钠(MIA)法制备KOA大鼠模型,随机分为模型组、贝母素乙(3.5 mg/kg)组、贝母素乙(3.5 mg/kg)+LPA(RhoA激活剂,1 mg/kg)组,每组 10 只,另选 10 只大鼠为对照组.机械刺激法和热辐射法检测大鼠膝关节疼痛症状;检测大鼠关节肿胀度、膝关节宽度并以步态评分评测其关节功能;透射电镜检测大鼠膝关节软骨细胞超微结构;TUNEL染色检测大鼠关节软骨细胞凋亡比;酶联免疫吸附(ELISA)法检测大鼠血清和关节液促炎因子白细胞介素(IL)-18、单核细胞趋化蛋白 1(MCP-1)水平;免疫印迹检测各组大鼠关节软骨组织凋亡(Cleaved caspase-3、Bax)和RhoA/ROCK通路相关蛋白表达.结果 贝母素乙可降低KOA大鼠关节肿胀度、膝关节宽度、步态评分、软骨细胞凋亡比、血清及关节液IL-18、MCP-1水平、软骨组织Cleaved caspase-3、Bax、RhoA、ROCK1、ROCK1 蛋白表达,升高机械痛阈值、热痛阈值(P<0.05);LPA可减弱贝母素乙对KOA大鼠关节软骨损伤的改善作用.结论 贝母素乙可通过抑制RhoA/ROCK信号激活而抑制KOA大鼠关节炎症反应和关节软骨细胞凋亡,减轻软骨损伤.
Abstract
Objective To investigate the effect of Peiminine on cartilage damage in knee osteoarthritis(KOA)rats by regulating the Ras homologous gene family,member A(RhoA)/Rho associated coiled coil-forming kinase(ROCK)signaling pathway.Methods A KOA rat model was prepared using the sodium iodoacetate(MIA)method and randomly grouped into a model group,a Peiminine(3.5 mg/kg)group,and a Peiminine(3.5 mg/kg)+LPA(RhoA activator,1 mg/kg)group,with 10 rats in each group,another 10 rats were selected as control group.Mechanical stimulation and thermal radiation method were applied to detect knee joint pain symptoms in rats;the joint swelling degree and knee joint width of rats were measured,and their joint function was evaluated using gait scores;transmission electron microscopy was applied to detect the ultrastructure of knee joint chondrocytes in rats;TUNEL staining was applied to detect the apoptosis ratio of articular chondrocytes;enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of pro-inflammatory factors such as interleukin-18(IL-18)and monocyte chemotactic protein-1(MCP-1)in rat serum and joint fluid;immunoblotting was applied to detect apoptosis(Cleaved caspase-3,Bax)and the expression of RhoA/ROCK pathway related proteins in the articular cartilage tissue of rats.Results Peiminine decreased joint swelling,knee joint width,gait score,chondrocyte apoptosis ratio,serum and joint fluid IL-18,MCP-1 levels,and cartilage Cleaved caspase-3,Bax,RhoA,ROCK1,ROCK1 protein expression in KOA rats,and increased mechanical pain threshold and heat pain threshold(P<0.05);LPA could weaken the amelioration effect of Peiminine on articular cartilage injury in KOA rats.Conclusion Peiminine can inhibit joint inflammation and apoptosis of articular chondrocytes in KOA rats by inhibiting RhoA/ROCK signal activation,and alleviate cartilage injury.
基金项目
湖北省武汉市卫生健康委员会科研项目(WZ22C63)
NETL
NSTL
万方数据