基于Nrf2-HO-1/CYP2E1通路探讨五指毛桃醇提取物对酒精性肝损伤小鼠的抗氧化保护机制  

Antioxidant Protective Mechanism of Alcohol Extract of Fici Hirtae Radix on Mice with Alcoholic Liver Injury Through Nrf2-HO-1/CYP2E1 Pathway

作　　者：张茹[1,2] 曲中原 杜娟[1] ZHANG Ru;QU Zhongyuan;DU Juan(College of Pharmacy,Harbin University of Commerce,Harbin 150076 Heilongjiang,China;Heilongjiang Provincial Key Laboratory of Drug Prevention and Treatment for Senile Diseases,Harbin 150076 Heilongjiang,China)

机构地区：[1]哈尔滨商业大学药学院,黑龙江哈尔滨150076 [2]黑龙江省预防与治疗老年病药物研究重点实验室,黑龙江哈尔滨150076

出　　处：《中药新药与临床药理》2021年第12期1769-1775,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：黑龙江省博士后资助项目(LBH-Z13128);哈尔滨商业大学青年创新人才培养计划项目(17XN034)。

摘　　要：目的探讨五指毛桃(Fici Hirtae Radix,FHR)醇提取物对小鼠酒精性肝损伤的保护作用及其机制。方法将60只小鼠随机分为对照组、模型组、阳性对照组及五指毛桃醇提取物高、中、低剂量组。除对照组给予生理盐水灌胃外,其余各组小鼠以梯度酒精灌胃法复制酒精性肝损伤模型。复制模型同时,阳性对照组灌胃给予50 mg·kg^(-1)水飞蓟素,五指毛桃醇提取物组分别灌胃给予2、1、0.5 g·kg^(-1)五指毛桃醇提取物进行干预,对照组、模型组灌胃给予等体积生理盐水,连续6周。复制模型成功后取材,HE染色法观察肝组织病理变化;比色法测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,肝组织内丙二醛(MDA)和还原型谷胱甘肽(GSH)含量,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性;并采用蛋白质印迹法测定核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、超氧化物歧化酶1(SOD-1)和细胞色素P450 2E1(CYP 2E1)的蛋白表达水平。结果与对照组比较,模型组小鼠肝细胞肿胀,排列散乱,中央静脉周围小变性明显,呈现肝细胞灶状坏死;血清中ALT和AST水平明显升高(P<0.01);肝组织中MDA含量明显升高(P<0.01),SOD活性明显降低(P<0.01),GSH含量和GSH-Px活性明显降低(P<0.01);Nrf2、HO-1及SOD-1蛋白水平明显降低(P<0.05),CYP2E1蛋白水平明显升高(P<0.05)。与模型组比较,五指毛桃醇提取物各剂量组肝脏炎性细胞浸润减少,肝脏组织坏死减轻;五指毛桃醇提取物高、中剂量组明显降低ALT、AST水平(P<0.05);五指毛桃醇提取物高、中剂量组明显降低MDA水平(P<0.05),五指毛桃醇提取物低剂量组明显升高GSH含量(P<0.05),五指毛桃醇提取物各剂量组均明显升高SOD水平(P<0.05),五指毛桃醇提取物高、低剂量组明显升高GSH-Px活性(P<0.05);五指毛桃醇提取物高剂量组明显上调Nrf2蛋白表达水平(P<0.05),五指毛桃醇提取物高、低剂量组明显上调HO-1Objective To investigate the protective effect and mechanism of alcohol extract of Fici Hirtae Radix(FHR)on alcoholic liver injury in mice.Methods Sixty mice were randomly divided into control group,model group,positive control group and FHR high-,medium-and low-dose groups.The control group was given normal saline by gavage,while the mice in other groups were given gradient alcohol by gavage to replicate the alcoholic liver injury model.And then,the positive control group was given 50 mg·kg^(-1) silymarin by gavage,the drug group was given 2 g·kg^(-1)、1 g·kg^(-1) and 0.5 g·kg^(-1) FHR by gavage,the control group and the model group were treated with equal volume of normal salin by gavage for 6 weeks,respectively.After the model was successfully made,the pathological changes of liver tissue were observed by HE staining,and the levels of ALT,AST,MDA,GSH,SOD,GSH-Px in liver tissue were measured.The protein expression levels of Nrf2,HO-1,SOD-1 and CYP2 E1 in liver tissue were detected by Western blot.Results Compared with the control group,the hepatocytes in the model group were swollen and scattered,the small degeneration around the central vein was significant.It was found that focal necrosis of hepatocytes,the levels of ALT and AST increased significantly(P<0.01),the level of MDA in liver tissue increased significantly(P<0.01),the activity of SOD decreased significantly(P<0.01),the content of GSH and activity of GSH-Px decreased significantly(P<0.05),the protein expression levels of Nrf2,HO-1 and SOD-1 decreased significantly(P<0.05),and the protein level of CYP2 E1 increased significantly(P<0.05).Compared with the model group,the infiltration of inflammatory cells and necrosis of liver tissue were reduced in FHR groups,the levels of ALT and AST in FHR high-,medium-dose groups were decreased(P<0.05),while the level of MDA was decreased obviously(P<0.05).The content of GSH in FHR low-dose group was significantly increased(P<0.05),the activities of SOD in FHR groups and GSH-Px in high-,low-dose groups were in

关 键 词：五指毛桃 醇提取物 酒精性肝损伤 氧化应激 Nrf2-HO-1/CYP2E1通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]