人参皂苷Rb3通过上调Klf16/Nrf2表达改善小鼠脂性肝细胞模型的脂质沉积与氧化应激  

Ginsenoside Rb3 Improves Lipidosis and Oxidative Stress in a Mouse Model of Lipidic Hepatocytes through Upregulation of Klf16/Nrf2 Expression

作　　者：吴肖婷 吴肖婕 崔晓丽 李湘 张东 WU Xiaoting;WU Xiaojie;CUI Xiaoli;LI Xiang;ZHANG Dong(The Third Operating Room of Binzhou People's Hospital,Binzhou 256600 Shandong,China;The Central Laboratory of Binzhou People's Hospital,Binzhou 256600 Shandong,China;Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006 Guangdong,China;Dept,of Gastroenterology,Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033 Guangdong,China)

机构地区：[1]滨州市人民医院住院三部手术室,山东滨州256600 [2]滨州市人民医院中心实验室,山东滨州256600 [3]广州中医药大学,广东广州510006 [4]深圳市中医院脾胃科,广东深圳518033

出　　处：《中药新药与临床药理》2022年第4期477-483,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：山东省重点研发计划项目(2019GSF108270)。

摘　　要：目的探讨人参皂苷Rb3对小鼠脂性肝细胞模型脂质沉积和氧化应激的影响。方法采用原位二步灌流法提取小鼠肝原代细胞,以油酸(OA)和棕榈酸(PA)混合培养基制备脂性肝细胞模型,并以人参皂苷Rb3低、中、高剂量(5、10、20 nmol·L^(-1))分别干预24 h。测定各组细胞中甘油三酯(TG)的含量;分别采用荧光探针BODIPY 493/503、BODIPY 581/591、DCFH-DA检测各组细胞脂质蓄积、脂质过氧化情况及活性氧(ROS)水平;采用qPCR法检测细胞Klf16、Nrf2 mRNA表达水平;Western Blot法检测细胞Klf16、Nrf2、Sod2蛋白表达水平。结果与正常组比较,模型组细胞中的TG含量显著升高(P<0.01),脂质蓄积和过氧化水平显著增强(P<0.05),ROS水平显著升高(P<0.01),Klf16、Nrf2 mRNA表达水平显著降低(P<0.01),Klf16、Nrf2及Sod2蛋白表达水平明显降低(P<0.05,P<0.01)。与模型组比较,人参皂苷Rb3高剂量组细胞中的TG含量明显降低(P<0.01),细胞脂质蓄积、过氧化水平明显降低(P<0.05),ROS水平显著降低(P<0.01),Klf16、Nrf2 mRNA表达水平明显升高(P<0.05,P<0.01),Klf16、Nrf2及Sod2蛋白表达水平明显升高(P<0.05,P<0.01)。结论人参皂苷Rb3可一定程度改善小鼠脂性肝细胞模型的脂质蓄积和氧化应激反应,其作用机制可能与上调Klf16/Nrf2表达有关。Objective To investigate the effects of Ginsenoside Rb3 on lipiddosis and oxidative stress in a mouse model of lipidic hepatocytes. Methods The mouse liver primary cells were extracted by in Situ two-step irrigation method,and the lipid liver primary cell model was prepared with mixed medium of oleic acid(OA)and palmitic acid(PA),and intervened with Ginsenoside Rb3 at low-,medium-and high-doses(5,10 and 20 nmol·L^(-1)) for24-hour. The triglyceride(TG) content of each group of cells was measured;the lipid accumulation, lipid peroxidation and ROS levels of each group of cells were detected by fluorescent probes BODIPY 493/503,BODIPY 581/591 and DCFH-DA,respectively. The expression levels of Klf16 and Nrf2 mRNA were detected by qPCR and the expression levels of Klf16,Nrf2 and Sod2 were detected by Western Blot method. Results Compared with the normal group,the TG content in model group was significantly increased(P<0.01),the lipid accumulation and lipid peroxidation level in the model group were significantly strengthened(P<0.05),and the fluorescence intensity of ROS(oxidative stress response)level was significantly increased;The mRNA expression levels of Klf16 and Nrf2 were significantly decreased(P<0.01),and the protein expression levels of Klf16,Nrf2 and Sod2 were significantly decreased(P<0.05,P<0.01). Compared with model group,the content of TG,lipid accumulation,peroxide and ROS in Ginsenoside Rb3 high-dose group were significantly decreased(P<0.05),and the mRNA expression levels of Klf16 and Nrf2 were significantly increased(P<0.05,P<0.01),the protein expression levels of Klf16,Nrf2 and Sod2 were significantly increased(P<0.05,P<0.01). Conclusion Ginsenoside Rb3 is able to improve lipid accumulation and oxidative stress in a mouse model of lipidic hepatocytes to some extent, and its mechanism of action may be related to the upregulation of Klf16 and Nrf2 mRNA and protein expression.

关 键 词：人参皂苷RB3 代谢相关脂肪性肝病 脂性肝细胞模型 脂质沉积 氧化应激 小鼠 

分 类 号：R285.5[医药卫生—中药学]