银杏叶片调控Nrf2-Keap1-ARE信号通路改善燃煤污染型地方性砷中毒大鼠肝损伤的作用  

Ginkgo biloba regulate Nrf2-Keap1-ARE signaling pathway to ameliorate liver injury in endemic arsenic poisoning rats induced by coal burning

作　　者：胡勇[1] 姚茂琳[1] 于春[1] 张爱华[1] Hu Yong;Yao Maolin;Yu Chun;Zhang Aihua(Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education,Department of Toxicology,School of Public Health,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学环境污染与疾病监控教育部重点实验室,公共卫生学院卫生毒理学系,贵阳550025

出　　处：《中华地方病学杂志》2021年第6期441-447,共7页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81430077、U1812403)。

摘　　要：目的探讨银杏叶片调控核因子E2相关因子2(Nrf2)-Kelch样ECH联合蛋白1(Keap1)-抗氧化反应元件(ARE)信号通路在燃煤污染型地方性砷中毒(简称燃煤型砷中毒)大鼠肝损伤中的作用。方法采用成组设计,30只Wistar大鼠按体重(80~100 g)采用随机数字表法分为5组,每组6只,雌雄各半。正常对照组常规喂饲4.5个月;银杏叶片对照组常规喂饲3个月后给予银杏叶片(25 mg/kg,6 d/周)1.5个月;饮水型砷中毒组和砷粮食污染组分别给予100 mg/L三氧化二砷(As_(2)O_(3))水溶液和含砷100 mg/kg饲料3个月后,常规喂饲1.5个月;银杏叶片治疗组喂饲含砷100 mg/kg饲料3个月后给予银杏叶片(25 mg/kg,6 d/周)1.5个月。采用硫代巴比妥酸比色法检测5组大鼠血清丙二醛(MDA)含量,黄嘌呤氧化酶法检测血清铜锌超氧化物歧化酶(SOD1)活力,二巯双硝基苯甲酸还原法检测血清谷胱甘肽过氧化物酶(GPx)活力;荧光定量PCR、免疫组织化学及免疫印迹法检测5组大鼠肝组织Nrf2-Keap1-ARE信号通路指标基因的mRNA和蛋白表达;并采用Pearson相关分析指标间的相关性。结果饮水型砷中毒组、砷粮食污染组和银杏叶片治疗组血清MDA含量[(3.54±0.51)、(3.83±0.87)、(2.93±0.84)μmol/L]均高于正常对照组[(1.85±0.36)μmol/L,P均<0.05];SOD1[(68.21±4.37)、(64.53±9.96)、(73.09±5.43)U/ml]和GPx活力[(486.41±40.45)、(458.24±42.25)、(539.79±79.43)U/L]均低于正常对照组[(81.47±5.73)U/ml、(747.86±80.33)U/L,P均<0.05];与砷粮食污染组比较,银杏叶片治疗组MDA含量较低,SOD1和GPx活力均较高(P均<0.05)。饮水型砷中毒组、砷粮食污染组和银杏叶片治疗组肝组织SOD1和GPx1 mRNA表达均高于正常对照组(P均<0.05),银杏叶片治疗组均高于砷粮食污染组(P均<0.05)。砷粮食污染组肝组织SOD1蛋白表达低于正常对照组(P<0.05),饮水型砷中毒组、砷粮食污染组和银杏叶片治疗组肝组织GPx1蛋白表达均低于正常对照组(P均<0.05)Objective To explore the effects of Ginkgo biloba on regulating NF-E2-related factor 2(Nrf2)-Kelch-like ECH-associated protein 1(Keap1)-antioxidant response element(ARE)signaling pathway in liver injury induced by coal-burning-borne endemic arsenic poisoning in rats.Methods Group design method was adopted,according to body weight(80-100 g),a total of 30 Wistar rats were divided into 5 groups(6 rats in each group,half males and half females)by random number table method.The normal control group was fed with normal diet ad libitum for 4.5 months;the Ginkgo biloba control group was fed with Ginkgo biloba(25 mg/kg,6 d/week)for 1.5 months after normal feeding for 3 months;the drinking water arsenic poisoning group and the arsenic contaminated grain group were fed respectively with 100 mg/L arsenic trioxide(As_(2)O_(3))solution and 100 mg/kg arsenic-containing feed for 3 months,and then fed with normal diet for 1.5 months;the Ginkgo biloba treatment group was fed with 100 mg/kg arsenic-containing feed for 3 months,and then was given Ginkgo biloba(25 mg/kg,6 d/week)for 1.5 months.After sacrificing the animals,the content of malondialdehyde(MDA),the activity of copper zinc superoxide dismutase(SOD1)and the activity of glutathione peroxidase(GPx)in serum were detected by thiobarbituric acid colorimetry,xanthine oxidase method and dimercaptodinitrobenzoic acid reduction method,respectively.The mRNA and protein expressions of indicator genes of Nrf2-Keap1-ARE signaling pathway in liver tissues were detected by quantitative real-time PCR,immunohistochemistry and Western blotting.Correlation between the indexes was analyzed by Pearson.Results In drinking water arsenic poisoning group,arsenic contaminated grain group and Ginkgo biloba treatment group,the contents of MDA in serum were(3.54±0.51),(3.83±0.87)and(2.93±0.84)μmol/L,respectively,which were higher than that in normal control group[(1.85±0.36)μmol/L,P<0.05];and SOD1 activities[(68.21±4.37),(64.53±9.96),(73.09±5.43)U/ml]and GPx activities[(486.41±40.45),(458.2

关 键 词：砷中毒 银杏叶片 氧化应激 肝损伤 核因子E2相关因子2 Kelch样ECH联合蛋白1 

分 类 号：R285.5[医药卫生—中药学]