三味干姜散对慢性肝损伤大鼠Nrf2、Bach1表达的影响

Effects of Sanwei Ganjiang Powder on Expression of Nrf2/Bach1 Factors in Rats with Chronic Liver Injury

目的研究三味干姜散对慢性肝损伤大鼠核因子NF-E2相关因子(Nrf2)、BTB-CNC同源体1(Bach1)表达的影响,初步探讨三味干姜散调节机体氧化应激抗肝损伤的作用机制。方法以四氯化碳(CCl4)诱导慢性肝损伤大鼠模型,灌胃给予三味干姜散(660 mg·kg-1)6周,测定大鼠肝脏、脾脏和胸腺指数;苏木精-伊红(HE)染色法观察肝脏病理变化;检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST),肝组织匀浆中丙二醛(MDA)含量和谷胱甘肽过氧化物酶(GSH-Px)活性;Western-Blot法测检Nrf2、Bach1、细胞核内与胞浆蛋白、血红素氧合酶1(HO-1)总蛋白;实时RT-PCR法检测Nrf2、Bach1和HO-1 mRNA水平。结果三味干姜散能改善CCL4所致肝损伤的病理表现,降低血清ALT、AST水平和肝组织匀浆MDA水平,提升GSH-PX活性(P<0.01,P<0.05);提高Nrf2核转位率与核Nrf2/Bach1比值,提高Nrf2的蛋白及mRNA的表达,降低Bach1蛋白及mRNA的表达,并使HO-1 mRNA及总蛋白水平提高(P<0.01)。结论三味干姜散可通过改变Nrf2核转位率与核Nrf2/Bach1比值调控Nrf2与Bach1在肝细胞中的表达,促进机体氧化应激的调节与抗肝损伤。

Objective To investigate the effects of Sanwei Ganjiang Powder on the expression of Nrf2/Bachl factors in rats with chronic liver injury, and to explore the mechanism of Sanwei Ganjiang Powder in regulating oxidative stress for anti-hepatic injury. Methods The rat model with chronic liver injury was induced by carbon tetrachloride（CC14） and given Sanwei Ganjiang Powder. Liver, spleen and thymus gland indices of rats were measured. Heptatic pathology was observed after hematoxylin-eosin staining （HE staining）. Serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase（AST）, malondialdehyde（MDA） content in hepatic homogenate and glutathione peroxidase （GSH-Px） activity were measured. Nrf2, Bachl, intranuclear and intraeytoplasm protein, and heine oxygenase 1 （HO-1） protein were measured by Western blotting method. Nrf2, Bachland HO-1 mRNA expression levels were measured by RT-PCR. Results After treatment with Sanwei Ganjiang Powder, CC14-indnced liver injury was improved. The serum ALT and AST levels and MDA content were reduced, and GSH-Px activity was increased （P〈 0.01）. Furthermore, nuclear translocation rate of Nrf2 and Nrf2/Bachl ratio were increased, proteins and mRNA levels of Nrf2 and HO-1 were increased, but roteins and mRNA levels of Bachl were reduced （P 〈 0.01）. Conclusion By increasing nuclear translocation rate of Nrf2 and Nrf2/Bachl ratio, Sanwei Ganjiang Powder has an effect on regulating the expression of Nrf2 and Bachl proteins in the liver cells and on improving oxidative stress and counteracting hepatic fibrosis.