益气活血方诱导破裂型椎间盘突出髓核细胞焦亡的机制研究  

Yiqi Huoxue Prescription Promotes Pyroptosis of Nucleus Pulposus Cells in Ruptured Disc Herniation

作　　者：王志强[1] 戴宇祥 林顺 朱宇[1] 许苏梁 俞鹏飞[1] 姜宏[1] 刘锦涛[1] 李晓春 WANG Zhiqiang;DAI Yuxiang;LIN Shun;ZHU Yu;XU Suliang;YU Pengfei;JIANG Hong;LIU Jintao;LI Xiaochun(Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Traditional Chinese Medicine,Suzhou 215000,Jiangsu China)

机构地区：[1]南京中医药大学附属苏州市中医医院(江苏苏州,215000)

出　　处：《中国中医骨伤科杂志》2025年第3期1-6,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家自然科学基金项目(82374220,82074467);江苏省中医药科技发展项目(MS2022080);第三批江苏省中医药领军人才项目(苏中医科教[2023]9号);江苏省科教能力提升工程:江苏省医学重点学科/实验室建设单位(苏卫科教[2022]17号);中医骨伤重点实验室建设项目(SZS2022019);苏州市科技发展计划项目(SKYD2023151,SKYD2023152);苏州市“科教兴卫”青年科技项目(KJXW2021047)。

摘　　要：目的:探究益气活血方对破裂型椎间盘突出髓核细胞的影响。方法:用益气活血方灌胃3个月龄SD大鼠制备含药血清。手术采集并分离破裂型腰椎间盘突出症患者的突出椎间盘组织,提取原代髓核细胞,分别采用正常血清和益气活血方含药血清干预。用ELISA法检测细胞上清中炎症因子IL-1β、IL-6和TNF-α含量变化;用CCK-8法检测细胞活力变化;用TUNEL染色检测细胞凋亡;用Western Blot法检测GSDMD-N、NLRP3、Cleaved-caspase1、p-NF-κB p65、IκBα表达情况。此外,进一步将益气活血方含药血清与NF-κB信号通路抑制剂联用,观察上述指标的变化情况。结果:益气活血方含药血清处理显著抑制髓核细胞增殖能力,差异有统计学意义(P<0.05)、细胞凋亡显著增加,差异有统计学意义(P<0.05)。益气活血方含药血清处理组IL-1β、IL-6和TNF-α含量显著升高,差异有统计学意义(P<0.05)。同时,益气活血方含药血清组细胞中GSDMD-N、NLRP3、Cleaved-caspase1、p-NF-κB p65的表达显著升高,差异有统计学意义(P<0.05),IκBα的表达水平显著降低,差异有统计学意义(P<0.01)。联用NF-κB信号通路抑制剂后这些指标的变化被抑制。结论:益气活血方能够通过调节NF-κB/NRLP3信号通路促进破裂型椎间盘突出髓核细胞焦亡。Objective:The purpose of this study is to explore the effects of Yiqi Huoxue prescription on nucleus pulposus cells of ruptured lumbar disc herniation.Methods:The Yiqi Huoxue prescription was orally administered to 3-month-old SD rats to prepare medicated serum.Surgical samples of protruding intervertebral disc tissue from patients with ruptured lumbar disc herniation were collected and nucleus pulposus cells were extracted for intervention with normal serum and Yiqi Huoxue prescription medicated serum.ELISA was used to detect changes in the levels of inflammatory factors IL-1β,IL-6,and TNF-αin the cell supernatant;CCK-8 assay was used to measure changes in cell viability;TUNEL staining was performed to detect cell apoptosis;Western Blot was conducted to analyze the expression of GSDMD-N,NLRP3,Cleaved-caspase1,p-NF-κB p65,and IκBα.Additionally,Yiqi Huoxue prescription medicated serum was combined with NF-κB signaling pathway inhibitors to observe the changes in the above indicators.Results:Treatment with Yiqi Huoxue prescription medicated serum significantly inhibited the proliferation capacity of nucleus pulposus cells(P<0.05)and significantly increased cell apoptosis(P<0.05).The levels of IL-1β,IL-6,and TNF-αwere significantly elevated in the Yiqi Huoxue prescription medicated serum treatment group(P<0.05).Moreover,the expression of GSDMD-N,NLRP3,Cleaved-caspase1,p-NF-κB p65 in cells treated with Yiqi Huoxue prescription medicated serum significantly increased(all P<0.05),while the expression level of IκBαsignificantly decreased(P<0.01).The changes in these indicators were suppressed when the NF-κB signaling pathway inhibitor was used in combination.Conclusion:Yiqi Huoxue prescription can promote apoptosis of nucleus pulposus cells in ruptured intervertebral disc herniation by regulating the NF-κB/NRLP3 signal.

关 键 词：益气活血方 腰椎间盘突出 髓核细胞 焦亡 

分 类 号：R-33[医药卫生]