基于JNK/p38 MAPK/NF-κB信号通路探究八桂止痛散对脂多糖诱导脊髓星形胶质细胞凋亡的影响  

The impact of Bagui Zhitong powder on LPS-induced apoptosis of spinal cord astrocytes based on the JNK/p38 MAPK/NF-κB signaling pathway

作　　者：廖承成 尹瑞文 张洪卫 张爽 李霞 彭国瑶 聂欣怡 LIAO Chengcheng;YIN Ruiwen;ZHANG Hongwei;ZHANG Shuang;LI Xia;PENG Guoyao;NIE Xinyi(Department of Dermatology,the First Clinical Medical College of Yunnan University of Chinese Medicine,Kunming 650021,China)

机构地区：[1]云南中医药大学第一临床医学院皮肤科,云南昆明650021 [2]云南中医药大学第一临床医学院脑病科,云南昆明650021

出　　处：《陕西中医》2024年第4期457-462,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：云南省科技厅科技计划项目(202001AZ070001-053);云南省教育厅科学研究基金资助项目(2023Y0488);云南省冯素英专家工作站建设项目(202105AF150035);云南省中医医院卓越人才项目[云中附党字(2021)5号]。

摘　　要：目的:基于JNK/p38 MAPK/NF-κB信号通路探究八桂止痛散对脂多糖(LPS)诱导的脊髓星形胶质细胞凋亡的影响。方法:从SD乳鼠脊髓中分离星形胶质细胞并鉴定。CCK-8法检测不同浓度八桂止痛散(2.5、5、10、20、40、80 mg/ml)对细胞存活的影响;随后分别设置对照组、LPS组、LPS+药物-L组、LPS+药物-M组、LPS+药物-H组、LPS+药物-H+Anisomycin组。CCK-8法、流式细胞术检测细胞存活、凋亡;ELISA法检测细胞中白介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)水平;qRT-PCR检测细胞中JNK、p38 MAPK、NF-κB p65 mRNA表达水平;Western blot检测JNK/p38 MAPK/NF-κB通路相关蛋白表达水平。结果:与对照组相比,LPS组细胞存活率显著降低,细胞凋亡率、IL-1β、IL-6、TNF-α水平、JNK mRNA、p38 MAPK mRNA、NF-κB p65 mRNA表达、p-JNK/JNK、p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65表达显著增加(均P<0.05);与LPS组相比,LPS+药物-L组、LPS+药物-M组、LPS+药物-H组细胞存活率显著增加,细胞凋亡率、IL-1β、IL-6、TNF-α水平、JNK mRNA、p38 MAPK mRNA、NF-κB p65 mRNA表达、p-JNK/JNK、p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65表达显著降低,具有剂量依赖性(均P<0.05);与LPS+药物-H组相比,LPS+药物-H+Anisomycin组细胞存活率显著降低,细胞凋亡率、IL-1β、IL-6、TNF-α水平、JNK mRNA、p38 MAPK mRNA、NF-κB p65 mRNA表达、p-JNK/JNK、p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65表达显著增加(均P<0.05)。结论:八桂止痛散可抑制LPS诱导的脊髓星形胶质细胞凋亡,其机制可能与抑制JNK/p38 MAPK/NF-κB信号通路有关。Objective:To explore the impact of Bagui Zhitong powder on LPS-induced apoptosis of spinal cord astrocytes based on the JNK/p38 MAPK/NF-κB signaling pathway.Methods:Astrocytes were isolated and identified from the spinal cord of SD suckling rats.The effects of different concentrations of Bagui Zhitong powder(2.5,5,10,20,40,80 mg/ml) on cell survival were detected by CCK-8 method.Subsequently,the control group,LPS group,LPS+drug low dose group,LPS+drug medium dose group,LPS+drug high dose group and LPS+drug high dose+Anisomycin group were set,respectively.CCK-8 and flow cytometry were applied to detect cell survival and apoptosis.The levels of interleukin-1β,IL-6 and tumor necrosis factor-α(TNF-α) were detected by ELISA.qRT-PCR was applied to detect the expression levels of JNK mRNA,p38 MAPK mRNA and NF-κB p65 mRNA in cells.Western blot was applied to detect the expression levels of JNK/p38 MAPK/NF-κB pathway related proteins.Results:Compared with the control group,the cell survival rate in the LPS group was obviously reduced,the apoptosis rate,IL-1β,IL-6,TNF-α levels,JNK mRNA,p38 MAPK mRNA,NF-κB p65 mRNA expression,p-JNK/JNK,p-p38 MAPK/p38 MAPK,p-NF-κB p65/NF-κB p65 expression were obviously increased(all P<0.05).Compared with the LPS group,the cell survival rate in the the LPS+drug L group,LPS+drug M group,and LPS+drug H group was obviously increased,the apoptosis rate,IL-1β,IL-6,TNF-α levels,JNK mRNA,p38 MAPK mRNA,NF-κB p65 mRNA expression,p-JNK/JNK,p-p38 MAPK/p38 MAPK,p-NF-κB p65/NF-κB p65 expression were obviously reduced,in a dose dependent manner(all P<0.05).Compared with the LPS+drug H group,the cell survival rate in the LPS+drug H+Anisomycin group was obviously reduced,the apoptosis rate,IL-1β,IL-6,TNF-α levels,JNK mRNA,p38 MAPK mRNA,NF-κB p65 mRNA expression,p-JNK/JNK,p-p38 MAPK/p38 MAPK,p-NF-κB p65/NF-κB p65 expression were obviously increased(all P<0.05).Conclusion:Bagui Zhitong powder can inhibit LPS-induced apoptosis of spinal astrocytes,which may be related to the inhibition of

关 键 词：神经病理性疼痛 八桂止痛散 脂多糖 脊髓星形胶质细胞 凋亡 神经炎症 

分 类 号：R285.5[医药卫生—中药学]