基于HMGB1/RAGE通路探讨银杏叶片对砷致大鼠肺损伤的改善作用  

Effects of Ginkgo biloba on arsenic-induced lung injury in rats based on HMGB1/RAGE pathway

作　　者：郑凡艳 王文娟[1] 张玉琼[2] 张爱华[1] Zheng Fanyan;Wang Wenjuan;Zhang Yuqiong;Zhang Aihua(Department of Toxicology,Key Laboratory of Environmental Pollution and Disease Control,Ministry of Education,School of Public Health,Guizhou Medical University,Guiyang 550025;Guizhou Center for Disease Control and Prevention,Guiyang 550004,China)

机构地区：[1]贵州医科大学公共卫生与健康学院毒理学系环境污染与疾病监控教育部重点实验室,贵阳550025 [2]贵州省疾病预防控制中心,贵阳550004

出　　处：《中华地方病学杂志》2022年第7期524-531,共8页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81430077、U1812403、81872569)。

摘　　要：目的:探讨银杏叶片对砷中毒大鼠肺损伤的拮抗与治疗作用及其机制。方法:共选择42只健康清洁级Wistar大鼠,雌雄各半,体重为120~130 g,按体重采用随机数字表法分为7组,每组6只。建立银杏叶片拮抗和银杏叶片治疗两种干预模型,具体处理如下:(1)银杏叶片拮抗实验研究(4组):对照A组给予去离子水;银杏叶片对照组(GBE组)给予银杏叶片溶液(50 mg·kg^(-1)·bw);砷中毒组(As组)给予亚砷酸钠(NaAsO_(2))溶液(10 mg·kg^(-1)·bw);银杏叶片拮抗组(As+GBE组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)处理的同时给予银杏叶片溶液(50 mg·kg^(-1)·bw),以上给药方式均为灌胃,每周6天,连续4个月。(2)银杏叶片治疗实验研究(3组):对照B组给予去离子水5.5个月;砷中毒自然恢复组(恢复组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)灌胃4.0个月后,用去离子水灌胃1.5个月;银杏叶片治疗组(治疗组)给予NaAsO_(2)溶液(10 mg·kg^(-1)·bw)灌胃4.0个月后,给予银杏叶片溶液(50 mg·kg^(-1)·bw)灌胃1.5个月,以上给药均为每周6天。采用Masson染色评价肺组织胶原纤维沉积情况;蛋白免疫印迹法(Western blot)检测肺组织匀浆中相关蛋白[炎性细胞因子基质金属蛋白酶(MMP)-9、白细胞介素(IL)-1β、IL-18,高迁移率族蛋白B1(HMGB1)/晚期糖基化终末产物受体(RAGE)通路的HMGB1、RAGE,磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)通路的PI3K、AKT、磷酸化AKT(p-AKT),转化生长因子(TGF)-β1/SMAD通路的TGF-β1、SMAD2、p-SMAD2、SMAD3、p-SMAD3、SMAD4]的表达水平。结果:(1)银杏叶片拮抗作用:与对照A组比较,GBE组大鼠肺组织各蛋白表达水平和胶原纤维沉积均未见明显变化(P均>0.05),As组大鼠肺组织MMP-9、IL-1β、IL-18蛋白表达水平和胶原纤维沉积均明显增加(P均<0.05),HMGB1、RAGE、PI3K、p-AKT、TGF-β1、p-SMAD2、p-SMAD3、SMAD4蛋白表达水平均明显升高(P均<0.05);与As组比较,As+GBE组大鼠肺组织MMP-9、IL-1β、IL-18蛋白Objective To investigate the antagonistic and therapeutic effects of Ginkgo biloba on arsenic-induced lung injury in rats and its mechanism.Methods A total of 42 healthy clean grade Wistar rats,half male and half female,weighing 120-130 g,were randomly divided into 7 groups with 6 rats in each group.Two intervention models of Ginkgo biloba antagonism and treatment were established,respectively.The specific treatments were as follows:(1)Experimental study on the antagonism of Ginkgo biloba(4 groups):the control A group was given deionized water;the Ginkgo biloba control(GBE)group was given Ginkgo biloba solution(50 mg·kg^(-1)·bw);the arsenic-treated(As)group was given sodium arsenite solution(10 mg·kg^(-1)·bw);the Ginkgo biloba antagonistic(As+GBE)group was treated with sodium arsenite solution(10 mg·kg^(-1)·bw)and Ginkgo biloba solution(50 mg·kg^(-1)·bw),and the above administration was by gavage for 6 days/week,for 4 months.(2)Experimental study on the treatment of Ginkgo biloba(3 groups):the control B group was given deionized water for 5.5 months;in the arsenism natural recovery(recovery)group,sodium arsenite solution(10 mg·kg^(-1)·bw)was administered by gavage for 4.0 months and deionized water for 1.5 months;the Ginkgo biloba treatment(treatment)group was given sodium arsenite solution(10 mg·kg^(-1)·bw)by gavage for 4.0 months and Ginkgo biloba solution(50 mg·kg^(-1)·bw)for 1.5 months,and the above administration was for 6 days/week.Masson staining was used to evaluate collagen fiber deposition in lung tissue.Western blotting was used to detect the expression level of related proteins in lung tissue homogenates,including inflammatory cytokines matrix metalloproteinase-9(MMP-9),interleukin(IL)-1β,IL-18;high mobility group box 1(HMGB1)and receptor for advanced glycation end-products(RAGE)of the HMGB1/RAGE pathway;phosphatidylinositol-4,5-bisphosphate 3-kinase(PI3K),protein kinase B(AKT),phosphorylated AKT(p-AKT)of the PI3K/AKT pathway;transforming growth factor(TGF)-β1,SMAD2,p-SMAD2,SMAD3,p-SMA

关 键 词：砷中毒 银杏叶片 肺 炎症 胶原纤维沉积 

分 类 号：R285.5[医药卫生—中药学]