青春期丙烯酰胺暴露对小鼠肺组织氧化损伤的影响  

Effects of adolescent exposure to acrylamide on oxidative injury of lung tissue in mice

作　　者：卜文婕[1] 林爱琴[1] 高继光[1] 潘涛 任悦鸣 孙敏[3] BU Wenjie;LIN Aiqin;GAO Jiguang;PAN Tao;REN Yueming;SUN Min(Department of Medical Biology of Wannan Medical College,Wuhu 241002,China;School of Public Health,Wannan Medical College,Wuhu 241002;School of Life Science,Anhui University,Hefei 230601)

机构地区：[1]皖南医学院医学生物学教研室,安徽芜湖241002 [2]皖南医学院公共卫生学院,安徽芜湖241002 [3]安徽大学生命科学学院,合肥230601

出　　处：《中国实验动物学报》2023年第1期43-50,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：安徽省属本科医学院校公共卫生协同创新项目(GXXT-2020-025);2021年度安徽高校自然科学研究重点项目(KJ2021A0823);2021年安徽省大学生创新创业训练计划(S202110368012)。

摘　　要：目的探究丙烯酰胺(acrylamide,ACR)对小鼠肺组织的损伤及其可能机制。方法以4周龄SPF级雄性小鼠为实验动物,随机分为对照组和ACR组(10 mg/(kg·d)组和20 mg/(kg·d)组),每组8只。对照组给予常规饮用水;ACR组按小鼠每日饮水5 mL来配制ACR的染毒浓度,每3 d更换1次,连续处理4周。实验结束后取肺组织制作切片,进行组织病理学分析;利用试剂盒检测其氧化损伤的相关指标;进一步利用免疫荧光与Western Blot技术检测Nrf2-ARE通路相关蛋白表达。结果与对照组相比,随ACR摄入浓度的增加,小鼠体重逐渐下降(P<0.01);组织病理学分析发现,ACR组小鼠肺组织出现细支气管上皮肥大,肺泡上皮增生及肺泡腔面积明显减少,20 mg/(kg·d)组病理学改变更为明显;氧化应激指标检测发现,与对照组相比,随着ACR摄入浓度的增加,肺组织中GSH-PX活性逐渐下降(P<0.05)、SOD活力逐渐下降(P<0.05)、CAT活力逐渐下降(P<0.01),而MDA的含量明显上升(P<0.01);免疫荧光结果显示,与对照组相比,ACR组肺组织中Nrf2蛋白表达显著上升,在10 mg/(kg·d)组表达最高(P<0.001),且出现核转位现象,其伴侣蛋白Keap1蛋白表达显著下降,在10 mg/(kg·d)组表达最低(P<0.001);Western Blot结果显示,与正常对照组相比,ACR组肺组织中Nrf2基因及下游抗氧化基因HO-1蛋白表达升高,在10 mg/(kg·d)组最高(P<0.01),伴侣蛋白Keap1表达显著下降,在10 mg/(kg·d)组表达最低(P<0.01),而基因NQO-1蛋白表达随着ACR摄入浓度增加逐渐升高(P<0.01)。结论青春期小鼠ACR暴露可导致肺组织损伤,该过程可能与肺组织氧化还原失衡有关。Objective To investigate the toxic effects of acrylamide(ACR)on the lung tissue of mice and its possible mechanism of action.Methods 4⁃week⁃old SPF male mice were randomly divided into a control and two ACR groups(10 mg/(kg·d)group and 20 mg/(kg·d)group),with eight mice in each group.The control group was given conventional drinking water.In the ACR groups,the exposure concentration of ACR was prepared according to 5 mL of sectioned for histopathological analysis.Related indexes of oxidative damage were detected.Immunofluorescence and Western Blot were used to detect the expression of Nrf2⁃ARE⁃pathway⁃related proteins.Results Compared with the control group,the body weight of mice decreased gradually with an increase in ACR concentration(P<0.01).Histopathological analysis showed that bronchiolar epithelial hypertrophy,alveolar epithelial hyperplasia,and the alveolar cavity area were significantly reduced in the ACR group,and the pathological changes were more obvious in the 20 mg/(kg·d)group.Compared with the control group,the activity of GSH⁃Px(P<0.05),SOD(P<0.05),and CAT(P<0.01)in lung tissue decreased with an increase in ACR concentration,while the content of MDA increased significantly(P<0.01).Immunofluorescence result showed that,compared with the control group,the ACR group’s expression of Nrf2 protein in the lung tissue was significantly increased,and the highest expression was found in the 10 mg/(kg·d)group(P<0.001).The nuclear translocation phenomenon occurred,and expression of the chaperone protein Keap1 was significantly decreased,with the lowest expression found in the 10 mg/(kg·d)group(P<0.001).Western Blot result showed that,compared with the normal control group,expression of the Nrf2 gene and its downstream antioxidant protein HO⁃1 was increased in the lung tissue of the ACR group,and the highest level was found in the 10 mg/(kg·d)(P<0.01).Expression of the chaperone protein Keap1 was significantly decreased,and the lowest expression was found in 10 mg/(kg·d)group(P<0.01),bu

关 键 词：丙烯酰胺 核因子红系2相关因子2 氧化损伤 

分 类 号：Q95-33[生物学—动物学]