胡芦巴碱对H_(2)O_(2)诱导L02肝细胞氧化损伤的保护作用及机制  

The protective effect of trigonelline on H_(2)O_(2)-induced oxidative injury of L02 hepatocytes and its mechanism

作　　者：王瑞楠 马宏婷 铁芳芳 胡娜[2] 王洪伦[2] 何彦峰 WANG Rui-nan;MA Hong-ting;TIE Fang-fang;HU Na;WANG Hong-lun;HE Yan-feng(College of Pharmacy,Qinghai Minzu University,Xining 810007,China;Key Laboratory of Tibetan Medicine Research,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810001,China)

机构地区：[1]青海民族大学药学院,青海西宁810007 [2]中国科学院西北高原生物研究所藏药研究重点实验室,青海西宁810001

出　　处：《中国药理学通报》2023年第2期294-299,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81960785);青海省基础研究计划项目(No 2020-ZJ-717)。

摘　　要：目的研究胡芦巴碱(trigonelline,TRG)对H_(2)O_(2)诱导的人肝细胞系L02氧化损伤的保护作用及机制。方法MTT法检测细胞活力;试剂盒法检测LDH、SOD、GSH、MDA和CAT的变化;Western blot法检测MAPK/Nrf2/HO-1通路和凋亡相关蛋白表达情况。结果650μmol·L^(-1) H_(2)O_(2)作用12 h可导致L02细胞出现明显损伤。TRG作用后可以显著提高损伤细胞的存活率,并且能浓度依赖性地降低LDH泄漏量和MDA的含量,增强GSH、SOD和CAT等酶的活性。Western blot结果显示,与模型组相比,TRG可以促进Nrf2和HO-1蛋白表达,上调Caspase 3/cleaved Caspase 3和Bcl-2/Bax水平,抑制p-JNK/JNK和p-ERK1/2/ERK1/2表达,从而对肝细胞起到保护作用。结论TRG保护H_(2)O_(2)诱导的L02细胞氧化损伤的作用机制可能与激活Nrf2/HO-1信号通路,抑制MAPK通路的表达,并通过提高Caspase 3/cleaved Caspase 3和Bcl-2/Bax表达比值改善细胞凋亡有关。Aim To study the protective effect of trigonelline on H_(2)O_(2)-induced oxidative damage of human liver cell line L02 and the underlying mechanism.Methods The cell viability was detected by MTT method;the changes of LDH,SOD,GSH,MDA and CAT were detected by kit method;the expression of MAPK/Nrf2/HO-1 pathway and apoptosis-related proteins were detected by Western blot.Results The treatment of 650μmol·L^(-1) H_(2)O_(2)for 12 h resulted in significant damage to L02 cells.TRG could significantly improve the survival rate of injured cells and reduce the leakage of LDH and the content of MDA in a concentration-dependent manner,and enhance the activities of enzymes such as GSH,SOD and CAT.Western blot results showed that compared with the model group,TRG could promote the expression of Nrf2 and HO-1 proteins,up-regulate the levels of Caspase 3/cleaved Caspase 3 and Bcl-2/Bax,and inhibit p-JNK/JNK and p-ERK1/2/ERK1/2 expression,thus protecting hepatocytes.Conclusions The mechanism of TRG protection against H_(2)O_(2)-induced oxidative damage in L02 cells may be related to activating the Nrf2/HO-1 signaling pathway,inhibiting the expression of MAPK pathway,and improving cell apoptosis by increasing the expression ratio of Caspase 3/cleaved Caspase 3 and Bcl2/Bax.

关 键 词：胡芦巴碱 L02细胞 氧化损伤 Nrf2/HO-1 细胞凋亡 MAPKS BCL-2/BAX 

分 类 号：R284.1[医药卫生—中药学] R322.47[医药卫生—中医学] R329.25R345.57R349.1