“易层”贴敷抑制NLRP3炎症小体激活改善膝骨关节炎的实验研究

Experimental research on the inhibition of NLRP3 inflammasome activation and improvement of knee osteoarthritis by Layers Adjusting External Application

目的基于NLRP3/Capase-1轴研究易层贴敷膏药改善KOA滑膜炎症的治疗机制。方法将SD大鼠分为正常组,KOA组,"易层"组,抑制剂组,各组大鼠、滑膜HE染色,Western Blot检测大鼠滑膜NLRP3、ASC、Caspase-1蛋白和软骨MMP-1、MMP-13蛋白表达水平,PCR检测滑膜NLRP3、ASC、Caspase-1 mRNA表达。结果 "易层"贴敷显著改善KOA大鼠滑膜炎症Krenn评分,IL-1β、TNF-α蛋白表达下调,差异具有统计学意义(P<0.05);在应用"易层"贴敷膏药后,KOA大鼠滑膜组织中NLRP3、ASC、Caspase-1蛋白及mRNA表达水平均较KOA组下调,差异具有统计学意义(P<0.05);而应用Caspase-1抑制剂后,具有同样的下调作用,两组间无统计学意义(P>0.05);应用"易层"贴敷膏药干预后,KOA大鼠软骨中MMP-1及MMP-13水平下调,差异具有统计学意义(P<0.05)。结论 "易层"贴敷膏药可能通过下调NLRP3及Caspase-1的表达抑制KOA滑膜炎症,减少软骨组织MMPs水平,发挥保护软骨的作用。

Objective To study the therapeutic mechanism of " Layers Adjusting External Application " plaster based on NLRP3/Capase-1 axis to improve KOA synovial inflammation. Methods SD rats were divided into normal group, KOA group, " Layers Adjusting External Application " group, inhibitor group, HE staining of synovium in each group, and rat NLRP3, ASC, Caspase-1 in synovium and MMP-1, MMP-13 protein expression levels in cartilage were detected by Western Blot. PCR detection of synovial MRP3,ASC,Caspase-1 mRNA expression. Results The "Layers Adjusting External Application " application significantly improved Krenn score, IL-1β and TNF-α protein down-regulation in KOA rats, and the difference was statistically significant(P<0.05). After applying "Layers Adjusting External Application" plaster,the expression levels of NLRP3, ASC, caspase-1 protein and mRNA in the synovial tissue of KOA rats were significantly lower than those in the KOA group(P<0.05). However, the same down-regulation was observed after the application of caspase-1 inhibitor. There was no significant difference between the two groups(P>0.05). After intervention with "Layers Adjusting External Application " plaster, the levels of MMP-1 and MMP-13 in KOA rat cartilage were down-regulated, the difference was statistically significant(P<0.05).Conclusion "Layers Adjusting External Application " plaster may inhibit the inflammation of KOA synovial membrane by down-regulating the expression of NLRP3 and caspase-1, reduce the level of MMPs in cartilage tissue, and play a role in protecting cartilage.