桂枝调节RIPK1/RIPK3信号通路干预KOA小鼠软骨坏死性凋亡  

Cassia twig regulates the RIPK1/RIPK3 signaling pathway and intervenes in necrotic apoptosis of cartilage in KOA mice

作　　者：苏子珊 陈拓 廖太阳 于利凯 田地 王培民[1] 张农山[1] SU Zishan;CHEN Tuo;LIAO Taiyang;YU Likai;TIAN Di;WANG Peimin;ZHANG Nongshan(Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,China;Key Laboratory of Traditional Chinese Medicine Research on Metabolic Diseases at Nanjing University of Traditional Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学附属医院/江苏省中医院,江苏南京210029 [2]南京中医药大学代谢病中医研究重点实验室,江苏南京210023

出　　处：《中国骨质疏松杂志》2024年第10期1420-1425,1465,共7页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金面上项目(82274545);江苏省医学重点学科/实验室建设单位项目(JSDW202252);江苏省中医院科主任学术提升专项(Y2022ZR26)。

摘　　要：目的 基于RIPK1/RIPK3信号通路探讨桂枝治疗膝骨关节炎(knee osteoarthritis, KOA)小鼠软骨细胞坏死性凋亡的作用机制。方法 选取48只雄性C57小鼠,随机分为空白组、造模组、Nec-1组[5 mg/(kg·d),腹腔注射给药]、桂枝低剂量组[0.43 g/(kg·d),灌胃给药]、桂枝中剂量组[0.65 g/(kg·d),灌胃给药]、桂枝高剂量组[2.6 g/(kg·d),灌胃给药]。除空白组外均采用内侧半月板失稳法构建KOA小鼠模型,经药物干预28 d后,取小鼠血清及软骨组织,采用HE染色及番红O-固绿染色观察小鼠膝关节组织病理形态;免疫荧光分析小鼠软骨RIPK3表现情况;使用qRT-PCR及Western Blot检测RIPK1、RIPK3、p-RIPK1、p-RIPK3等相关mRNA及蛋白表达;使用ELISA检测TNF-α、IL-1β、IL-6含量。结果 与正常组相比,造模组小鼠关节软骨边缘严重破坏,RIPK1、RIPK3、p-RIPK1、p-RIPK3的mRNA及蛋白表达明显升高(P<0.05),TNF-α、IL-1β、IL-6含量明显升高(P<0.05);桂枝低剂量、中剂量、高剂量组及Nec-1组软骨结构趋于正常,RIPK1、RIPK3、p-RIPK1、p-RIPK3的mRNA及蛋白表达明显降低(P<0.05),IL-1β、IL-6、TNF-α含量明显降低(P<0.05)。结论 桂枝可以通过调节RIPK1/RIPK3信号通路,抑制膝关节软骨细胞坏死性凋亡,从而改善小鼠膝关节炎症环境,延缓KOA病理进程。Objective To investigate the mechanism of cassia twig in treating the necroptosis of KOA mouse chondrocytes based on RIPK1/RIPK3 signaling pathway.Methods Forty-eight male C57 mice were randomly divided into normal group,model group,Nec-1 group(5 mg/(kg·d)),cassia twig low-dose group(0.43 g/(kg·d)),cassia twig medium-dose group(0.65 g/(kg·d))and cassia twig high-dose group(2.6 g/(kg·d)).The mouse model of KOA was established by medial meniscus instability method,and the corresponding intragastric administration was performed after the modeling.After 28 days of drug intervention,the serum and cartilage tissues of mice were taken,and the histopathologic morphology of the knee joint of mice was observed by HE staining and saffranine O-solid green staining.The expression of RIPK3 in mouse cartilage was analyzed by immunofluorescence.The expression of RIPK1,RIPK3,p-RIPK1,p-RIPK3,TNF-α,IL-1β,IL-6 and other relatedmRNA and proteins were detected by RT-qPCR,ELISA and Western Blot.Results Compared with the normal group,the articular cartilage edge of the model group mice was severely damaged,and the mRNA and protein expressions of RIPK1,RIPK3,p-RIPK1,and p-RIPK3 were significantly increased(P<0.05).The content of TNF-α,IL-1β,and IL-6 significantly increased(P<0.05).The cartilage structure of the low-dose,medium dose,high-dose,and Nec-1 groups of cassia twig tended to be normal,and the mRNA and protein expressions of RIPK1,RIPK3,p-RIPK1,and p-RIPK3 were significantly reduced(P<0.05).The content of IL-1β,IL-6 and TNF-αsignificantly decreased(P<0.05).Conclusion By regulating RIPK1/RIPK3 signaling pathway,cassia twig can inhibit the necroptosis of knee cartilage cells,thereby improving the inflammatory environment of the knee joint in mice and delaying the pathological process of KOA.

关 键 词：桂枝 软骨细胞 坏死性凋亡 膝骨关节炎 信号通路 

分 类 号：R-332[医药卫生] R684.3