基于cGAS-STING信号通路探讨川芎嗪抑制Ca2+浓度保护膝骨关节炎大鼠软骨的作用机制研究
Investigation of the mechanism of action of Tetramethylpyrazine in inhibiting concentration of Ca2+to protect cartilage in knee osteoarthritis rats based on the cGAS-STING pathway
摘要
Abstract
Objective:To investigate whether Tetramethylpyrazine(TMP)can inhibit Ca2+concentration through the cGAS-STING pathway and thus protect cartilage in KOA rats.Methods:The KOA model was replicated using the anterior cruci-ate ligament disarticulation method,and male SD rats were randomly divided into the control group,the model group,and the TMP group(20,40,and 80 mg∙kg-1∙d-1),with 8 rats in each group.Chondrocytes from the knee joints of male SD rats were ex-tracted and divided into the control group,the IL-1β group,the TMP group(100 and 200 μmol/L)and the H-151 group(STING pathway inhibitor).The KOA model was established by inducing chondrocytes with IL-1β for 24 h,and TMP and H-151 inter-vened for 24h.The effect of TMP on cell viability was detected by CCK-8 assay.Pathological changes in cartilage tissues and chondrocytes were observed using Senna-Solid Green staining.Detection of cGAS,STING protein expression in tissues was mea-sured by immunohistochemical methods.RT-qPCR was used to detect cGAS,STING,Calmodulin,Calpain、NF-κB mRNA levels in tissues.Cellular Ca2+concentration was measured by flow cytometry.Western blot was used to detect intracellular and tis-sue cGAS,STING,Calmodulin,Calpain protein expression and relative cellular p-IRF3 protein expression.Results:CCK-8 as-say showed that TMP had no significant effect on cell viability in the concentration range of 0-400 μmol/L.The results of Senna-Solid green staining after TMP intervention showed improved cartilage tissue damage in all groups compared to the model group.The proportion of cGAS and STING protein-positive areas of cartilage tissue detected by immunohistochemistry was signif-icantly reduced.The relative expression levels of cGAS,STING,Calmodulin,Calpain and NF-κB mRNA also showed a de-creasing trend.In addition,in vitro experiments also re-verified that the chondrocyte Ca2+concentration was reduced in the TMP group compared to the IL-1β group.The relative expression of cGAS,STING,Calmodulin,Calpain and p-IRF3 proteins was re-duced.Conclusion:TMP protects cartilage in KOA rats via the cGAS-STING pathway and thus inhibits Ca2+concentration.关键词
膝骨关节炎/软骨/cGAS-STING信号通路/Ca2+Key words
Knee osteoarthritis/Cartilage/cGAS-STING signalling pathway/Ca2+分类
医药卫生引用本文复制引用
施蕾,廖太阳,刘德仁,刘江宇,王培民,吴鹏,茆军..基于cGAS-STING信号通路探讨川芎嗪抑制Ca2+浓度保护膝骨关节炎大鼠软骨的作用机制研究[J].海南医科大学学报,2025,31(16):1201-1210,10.基金项目
This study was supported by the National Natural Science Foundation of China(82205143),the Excellent Young Doctor Program of Jiangsu Hospital of Chinese Medicine(2023QB0122),the Third Batch of Peak Academic Talents of Jiangsu Provincial Hospital of Traditional Chinese Medicine(y2021rc20),Clinical Medical Innovation Center for Knee Osteoarthritis,Jiangsu Province Hospital of Traditional Chinese Medicine(Y2023zx05),Jiangsu Provincial Medical Key Discipline(Laboratory)Cultivation Unit(JSDW202252),Provincial Department of Education Postgraduate Innovation Project(KYCX24_2227) 国家自然科学基金青年项目(82205143) （82205143）
江苏省中医院优秀青年博士培养专项(2023QB0122) （2023QB0122）
江苏省中医院第三批高峰学术人才(y2021rc20) （y2021rc20）
江苏省中医院中医膝骨关节炎临床医学创新中心(Y2023zx05) （Y2023zx05）
江苏省医学重点学科/实验室建设单位(JSDW202252) （JSDW202252）
江苏省教育厅研究生创新项目(KYCX24_2227) （KYCX24_2227）
