摘要
目的:探究赶黄草配伍葛花制备的含药血清对乙醇诱导的L-02肝细胞的影响及其可能机制。方法:分别制备不同配比含药血清后,将L-02肝细胞分为6组进行实验:空白对照组,模型组,赶黄草配伍葛花1∶1组、2∶1组和1∶2组,凯希莱组。MTT法检测含药血清对乙醇诱导的L-02细胞存活率的影响;酶标法检测培养液中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)及受损细胞中超氧化物歧化酶(SOD)和丙二醛(MDA)的水平;realtime PCR法和Western blot检测赶黄草配伍葛花对L-02肝细胞中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、核因子E2相关因子2(Nrf2)和血红素加氧酶1(HO-1)mRNA及蛋白表达的影响。结果:与空白对照组相比,模型组ALT、AST及MDA水平明显升高,SOD活性明显降低(P<0.01);与模型组相比,各药物组ALT、AST及MDA水平明显降低,SOD活性明显升高(P<0.01)。机理研究中,与空白对照组相比,模型组TNF-α和IL-6的mRNA和蛋白表达显著升高,Nrf2和HO-1的mRNA和蛋白表达显著降低(P<0.01);与模型组相比,各配伍组TNF-α和IL-6的mRNA和蛋白表达显著降低,Nrf2和HO-1的mRNA和蛋白表达显著升高(P<0.01)。结论:赶黄草配伍葛花含药血清可能通过下调TNF-α和IL-6的表达,上调Nrf2和HO-1的表达,从而降低L-02肝细胞的炎症反应并减轻氧化应激,进而发挥治疗酒精性肝损伤的作用。
AIM： To explore the effect of Penthorum chinense Pursh and Puerariae flos-containing serum on L-02 liver cell injury induced by alcohol and its possible mechanism. METHODS： After preparing drug-containing serum,the L-02 cells cultured in vitro were divided into 6 groups： blank control group,model group,1∶ 1 group,2∶ 1 group and1∶ 2 group of combination of Penthorum chinense Pursh and Puerariae flos,and tiopronin group. The viability of the L-02 cells was measured by MTT assay. The activity of alanine aminotransferase（ ALT）,aspartate aminotransferase（ AST） and superoxide dismutase（ SOD）,and the content of malondialdehyde（ MDA） were detected by enzyme label methods. The expression of tumor necrosis factor-α（ TNF-α）,interleukin-6（ IL-6）,nuclear factor E2-related factor 2（ Nrf2） and heme oxygenase-1（ HO-1） at mRNA and protein levels was determined by real-time PCR and Western blot,respectively. RESULTS： Compared with control group,the levels of ALT,AST and MDA were increased significantly,and SOD was decreased in model group（ P〈0. 01）. Compared with model group,these indexes in all treatment groups were opposite（ P〈0. 01）. Compared with control group,the expression of TNF-α and IL-6 at mRNA and protein levels was significantly increased,the mRNA and protein expression of Nrf2 and HO-1 was decreased（ P〈0. 01）. Compared with model group,these indexes in combination groups were opposite（ P〈0. 01）. CONCLUSION： The therapeutic effects of Pentehorum chinensa Pursh and Puerariae flos-containing serum may affect the expression levels of TNF-α,IL-6,Nrf2 and HO-1,and reduce the inflammatory reaction and oxidative stress in alcohol-induced L-02 liver cells,which plays a role in attenuating alcoholic liver injury.
作者
舒承倩
吴晶魁
周滢
SHU Cheng-qian;WU Jing-kui;ZHOU Ying(Traditional Chinese Meidicine Research Laboratory,College of Traditional Chinese Medicine,Chongqing Medical Universi ty,Chongqing 400016,China;Tianjin Meilical University,Tianjin 300070,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第6期1095-1100,共6页
Chinese Journal of Pathophysiology
基金
重庆市渝中区科技计划项目(渝中科2017-23号)
关键词
赶黄草
葛花
酒精性肝病
炎症
氧化应激
Penthorum chinense Pursh
Puerariae flos
Alcoholic liver disease
hfflammation
Oxidative
