小檗碱对棕榈酸诱导大鼠心肌细胞H9c2脂毒性损伤的影响  

Effect of Berberine on Palmitic Acid-induced Hyperlipidemia Injury in H9c2 Cardiomyocytes of Rats

作　　者：杨茹[1] 覃金红[2] 刘丹霞[2] 陈清杰[2] 吴宁华[2] Yang Ru;Qin Jinhong;Liu Danxia;Chen Qingjie;Wu Ninghua(Department of Pharmacy,Xiantao First People’s Hospital,Hubei Xiantao 433000,China;School of Basic Medicine,Hubei University of Science and Technology)

机构地区：[1]仙桃市第一人民医院药学部,湖北仙桃433000 [2]湖北科技学院医学部基础医学院

出　　处：《中国药师》2022年第10期1703-1707,共5页China Pharmacist

基　　金：湖北科技学院校内培育项目(编号:2020-21X30);湖北省教育厅科学研究计划资助重点项目(编号:D20202803)。

摘　　要：目的:探讨小檗碱对棕榈酸(PA)诱导大鼠心肌细胞H9c2脂毒性损伤的影响。方法:在正常培养的H9c2细胞中,采用细胞计数试剂盒(CCK8)检测细胞存活率;采用ELISA试剂盒检测细胞裂解液中氧化还原指标:丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH);采用免疫印迹法Western blot检测细胞内能量感受器磷酸化AMP依赖的蛋白激酶(AMPK)/AMPK及肝激酶B1(LKB1)、沉默调节蛋白1(SIRT1)和成纤维细胞生长因子-21(FGF-21)的蛋白表达。结果:PA在100μmol·L^(-1)浓度时,处理H9c2细胞24 h以上产生明显细胞毒性,与正常对照组(不含PA)比较,差异有统计学意义(P<0.05或P<0.01)。使用5μmol·L^(-1)小檗碱预处理H9c2细胞4 h可以逆转PA对H9c2中MDA、SOD和GSH的影响,同时细胞存活率明显上升,差异有统计学意义(P<0.05)。同时,在给予小檗碱治疗后,p-AMPK/AMPK、LKB1、SIRT 1和FGF-21蛋白的表达水平亦显著增加(P<0.05)。但使用FGF21的干扰RNA(siRNA)后,小檗碱激活AMPK的现象几乎被逆转,差异有统计学意义(P<0.05)。结论:小檗碱可显著改善PA诱导的心肌细胞脂毒性,其机制可能与经典的FGF-21/AMPK信号通路有关。Objective:To explore the protective effect of berberine on palmitic acid(PA)-induced lipotoxicity in H9 c2 cells.Methods:In the normal cultured H9 c2 cells,CCK8 assay was used to assess the cell survival rate;the values of MDA,SOD and glutathione(GSH)were detected by commercial ELISA kits;Western blot was used to detect the protein expression of p-AMPK/AMPK,LKB1,SIRT 1 and FGF-21.Results:The cytotoxicity effects of PA at the concentration of 100μmol·L^(-1) on H9 c2 cardiomyocytes were obvious after 24-hour treatment,and compared with normal control group(without PA)the difference was statistically significant(P<0.05 or P<0.01).Pretreatment of H9 c2 cells with 5μmol·L^(-1) berberine for 4 h could reverse the effects of PA on MDA,SOD and GSH in H9 c2,and the cell survival rate increased significantly(P<0.05).Meanwhile,after berberine treatment,the expression of p-AMPK/AMPK,LKB1,SIRT 1 and FGF-21 increased significantly(P<0.05).However,after using FGF21 interfering RNA(siRNA),the effect of berberine on activating AMPK was almost reversed,and the difference was statistically significant(P<0.05).Conclusion:Berberine can alleviate PA-induced cardiomyocytes lipotoxicity,and its mechanism is related to the classical FGF-21/AMPK signaling pathway.

关 键 词：小檗碱 心肌细胞H9c2 棕榈酸 AMP依赖的蛋白激酶 成纤维细胞生长因子-21 

分 类 号：R965.1[医药卫生—药理学]