基于TLR4/NF-κB/iNOS通路的截断逆挽方药物血清对L02细胞氧化应激损伤的影响  

Effects of Jieduan Niwan Prescription Drug Containing Serum on Oxidative Stress Injury of L02 Cells Based on TLR4/NF-κB/iNOS Pathway

作　　者：窦博 马重阳 侯伟欣 魏晓艺 姜雪娇 叶倩伶 胡文涵 张海天 张秋云[1] DOU Bo;MA Chongyang;HOU Weixin;WEI Xiaoyi;JIANG Xuejiao;YE Qianling;HU Wenhan;ZHANG Haitian;ZHANG Qiuyun(School of Traditional Chinese Medicine,Capital Medical University,Beijing Key Laboratory of Liver Diseases of Traditional Chinese Medicine,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院,中医肝病研究北京市重点实验室,北京100069

出　　处：《中国中医药信息杂志》2022年第4期87-91,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：北京市自然科学基金(7192024)。

摘　　要：目的观察截断逆挽方药物血清对D-氨基半乳糖(D-GaLN)诱导的L02细胞氧化应激损伤的影响,从TLR4/NF-κB/iNOS通路探讨其作用机制。方法体外培养L02细胞,D-GaLN诱导L02细胞损伤模型,将细胞分为正常组、模型组、截断逆挽方药物血清组、N-乙酰半胱氨酸组。CCK-8法检测细胞存活率,丙二醛(MDA)和谷胱甘肽(GSH)检测试剂盒检测细胞MDA和GSH含量,流式细胞术检测细胞活性氧(ROS)含量,Western blot检测Toll样受体4(TLR4)、核因子-κB p65(NF-κB p65)、一氧化氮合酶(iNOS)蛋白表达。结果与正常组比较,模型组L02细胞存活率明显降低(P<0.01),ROS、MDA含量明显增加,GSH含量明显减少,TLR4、NF-κB p65、iNOS蛋白表达明显升高,差异均有统计学意义(P<0.01);与模型组比较,截断逆挽方药物血清组L02细胞存活率明显升高(P<0.01),ROS、MDA含量明显减少,GSH含量明显增加,TLR4、NF-κB p65、iNOS蛋白表达明显降低,差异均有统计学意义(P<0.01)。结论截断逆挽方药物血清能抑制TLR4/NF-κB/iNOS通路介导的氧化应激,减轻D-GaLN诱导的L02细胞损伤。Objective To observe the effects of Jieduan Niwan Prescription drug containing serum on oxidative stress injury of L02 cells induced by D-galactosamine(D-GaLN);To explore the mechanism through TLR4/NF-κB/iNOS pathway.Methods L02 cells were cultured in vitro and modeled by D-GaLN.The cells were divided into normal group,model group,Jieduan Niwan Prescription drug containing serum group and N-acetylcysteine group.CCK-8 method was used to detect survival rate of L02 cells.The contents of MDA and GSH were detected by MDA and GSH test kits.The content of ROS was detected by flow cytometry.The expressions of TLR4,NF-κB p65,and iNOS protein were detected by Western blot.Results Compared with normal group,the survival rate of L02 cells in the model group significantly reduced(P<0.01),the contents of ROS and MDA significantly increased,the content of GSH significantly reduced,and the expressions of TLR4,NF-κB p65,and iNOS protein significantly increased,with statistical significance(P<0.01).Compared with the model group,the survival rate of L02 cells in the Jieduan Niwan Prescription serum containing drug group significantly increased(P<0.01),the contents of ROS and MDA significantly reduced,the content of GSH significantly increased,and the expressions of TLR4,NF-κB p65,and iNOS protein significantly decreased,with statistical significance(P<0.01).Conclusion Jieduan Niwan Prescription drug containing serum can inhibit TLR4/NF-κB/iNOS pathway mediated oxidative stress and reduce D-GaLN-induced L02 cell damage.

关 键 词：截断逆挽方 药物血清 氧化应激 慢加急性肝衰竭 TLR4/NF-κB/iNOS通路 L02细胞 

分 类 号：R285.5[医药卫生—中药学]