摘要
目的 基于NOD样受体热蛋白结构域相关蛋白3(NLRP3)/胱天蛋白酶-1(Caspase-1)/消皮素D(GSDMD)通路探讨重骨颗粒对膝骨关节炎大鼠细胞焦亡的作用机制.方法 随机将32只SPF级SD大鼠分为空白组、模型组、阳性对照组和重骨颗粒组,每组8只.空白组不作处理,其他组大鼠麻醉后用木瓜蛋白酶溶液注入关节腔进行膝骨关节炎造模1周,造模成功后各组干预治疗4周.电镜下观察膝关节软骨细胞焦亡情况,ELISA法检测血清中IL-18、IL-17、IL-6、IL-1β水平;免疫荧光测各组大鼠软骨组织中 NLRP3、Caspase-1、GSDMD、ASC 蛋白表达;qRT-PCR 法检测 NLRP3、Caspase-1、GSDMD mRNA 表达水平.结果 与模型组比较,重骨颗粒能抑制软骨组织的病理损伤,减少大鼠血清中IL-1β、IL-18、IL-6、IL-17表达(P＜0.05),降低软骨细胞焦亡相关因子NLRP3、Caspase-1、GSDMD mRNA和蛋白表达(P＜0.05).结论 重骨颗粒能够改善膝骨关节炎大鼠关节软骨组织损伤,降低炎症因子表达,改善细胞焦亡程度,其机制可能与调控NLRP3/Caspase-1/GSDMD信号通路减轻细胞焦亡有关.
Abstract
Objective To explore the mechanism of the effect of Chonggu granules on pyroptosis in rats with the knee osteoarthritis(KOA)through the NLRP3/Caspase-1/GSDMD pathway.Methods Thirty-two SPF SD rats were randomly assigned to four groups:Chonggu granule,model,blank,and positive control.Each group consisted of eight rats.Except for rats in the blank group,rats in other groups received injection of papain solution into the joint cavity under anesthesia for one week to achieve knee osteoarthritis model.Rats in each group received treatment for four weeks following the successful modeling.Under an electron microscope,the burned-out knee cartilage cells were examined.ELISA was used to measure the serum levels of IL-18,IL-17,IL-6,and IL-1β.Using immunofluorescence,the expressions of NLRP3,caspase-1,GSDMD,and ASC were quantified in the cartilage tissues of rats.Using qRT-PCR,the mRNA expression levels of GSDMD,caspase-1,and NLRP3 were examined.Results Compared to the model group,Chonggu granules were able to prevent pathological cartilage tissue damage and to decrease the serum expressions of IL-1β,IL-18,IL-6,and IL-17 in rats(P＜0.05).They also reduced the mRNA and protein expressions of GSDMD,Caspase-1,NLRP3(P＜0.05).Conclusion In KOA rats,articular cartilage damage can be improved with Chonggu granules,which also decrease the degree of apoptosis and the production of inflammatory markers.The mechanism behind these effects may be connected to the modulation of the NLRP3/Caspase-1/GSDMD signaling pathway,which lowers cell death.
基金项目
安徽省临床医学研究转化专项项目(202304295107020114)
安徽省临床医学研究转化专项项目(202304295107020115)
大健康研究院新安医学与中医药现代化研究所专项(2023CXMMTCM004)
大健康研究院新安医学与中医药现代化研究所专项(2023CXMMTCM015)
维普
万方数据