摘要
目的:检测小鼠睾丸细胞中肌醇需求酶1α(IRE1α)和X盒结合蛋白1(XBP1)mRNA和蛋白的表达,探讨IRE1-XBP1通路激活在低剂量辐射(LDR)诱导小鼠睾丸细胞内质网应激中的作用。方法:50只健康雄性ICR小鼠,随机分为10组,经75 mGy X射线全身照射后不同时间(0、3、6、12和24h)及不同剂量(0、50、75、100和200mGy)X射线照射后12h处死,分别采用实时定量PCR法和Western blotting法检测小鼠睾丸细胞中IRE1α、Total-XBP1(T-XBP1)和Spliced-XBP1(S-XBP1)mRNA表达水平及蛋白表达强度。结果:75mGy X射线照射后0~24h,小鼠睾丸细胞中IRE1α、T-XBP1和S-XBP1mRNA(除了3h时T-XBP1和S-XBP1mRNA)表达水平均随时间延长而升高,并在6h时达到峰值,而后逐渐降低;与0h组比较,6、12和24h组IRE1αmRNA、6和12h组T-XBP1mRNA及S-XBP1mRNA表达水平均明显升高(P<0.05或P<0.01)。与0h组比较,不同时间组IRE1α和S-XBP1蛋白表达强度升高,6和12h组IRE1α蛋白表达强度升高最明显,24h组S-XBP1蛋白表达强度升高最明显,而T-XBP1蛋白表达强度稍有降低。0~200mGy照射后12h,小鼠睾丸细胞中IRE1α、T-XBP1和S-XBP1mRNA表达水平升高,75mGy X射线照射后升高达峰值后逐渐降低,甚至降至0mGy以下;与0mGy组比较,75和200mGy组小鼠睾丸细胞中IRE1α、75mGy组小鼠睾丸细胞中T-XBP1和S-XBP1mRNA表达水平明显升高(P<0.05或P<0.01)。与0mGy组比较,75mGy组IRE1α和S-XBP1蛋白表达强度升高,75和200mGy组小鼠睾丸细胞中S-XBP1蛋白表达强度升高最明显,而T-XBP1蛋白表达强度无明显变化。结论:LDR可诱导小鼠睾丸细胞中IRE1α和S-XBP1mRNA表达水平和蛋白表达强度增加,从而激活内质网应激中的IRE1-XBP1信号通路。
Objective:To detect the expressions of inositol-requiring enzyme-1α(IRE1α)and X box-binding protein-1(XBP1)mRNA and proteins in mouse testis cells,and to explore the role of IRE1-XBP1 pathway activation in the endoplasmic reticulum stress induced by low dose radiation(LDR)in the mouse testis cells.Methods:A total of 50 heatly male ICR mice were randomly divided into 10 groups.After the mice were irradiated with 75 mGy X-ray in whole body at different time(0,3,6,12,and 24 h)or with different doses(0,50,75,100,and 200 mGy)of X-ray for12 h,the expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA and proteins were measured by real-time quantitative PCR and Western blotting method,respectively.Results:The expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA(except T-XBP1 and S-XBP1 mRNA at 3 h after irradiation)in the testis cells of the mice after irradiated with 75 mGy X-ray for 0-24 h were increased with the time prolongation,and reached to the maximum value at 6 h after irradiation,then were gradually decreased.Compared with 0 h group,the expression levels of IRE1αmRNA(6,12,and 24 h after irradiation),T-XBP1 mRNA and S-XBP1 mRNA(6 and 12 h after irradiation)were significantly increased(P<0.05 or P<0.01);the expression intensities of IRE1αand S-XBP1 proteins were increased;the expression intensities of IRE1α(6 and 12 h after irradiation)and S-XBP1(24 h after irradiation)proteins were increased most significantly,but the expression intensity of T-XBP1 protein was slightly decreased.The expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA in the testis cells of the mice after irradiated with 0-200 mGy for 12 h were increased,and reached to the maximum values after 75 mGy irradiation,then they were gradually decreased,even below 0 mGy.Compared with 0 mGy group,the expression levels of IRE1αmRNA in the testis cells of the mice in 75 and 200 mGy groups and the expression levels of T-XBP1 mRNA a nd S-XBP1 mRNA in 75 mGy group were significantly increased(P<0.05 or P<0.01);the expression intensities of IRE1αprotein
作者
于雷
唐庚
方芳
李鑫
龚守良
王志成
王剑锋
YU Lei;TANG Geng;FANG Fang;LI Xin;GONG Shouliang;WANG Zhicheng;WANG Jianfeng(Department of Radiotherapy,Second Hospital,Jilin University,Changchun 130041,China;Key Laboratory of Radiobiology,Ministry of Health,School of Public Health,Jilin University,Changchun 130021,China;Department of Radiotherapy,China-Japan Union Hospital,Jilin University,Changchun 130033,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2018年第5期919-923,共5页
Journal of Jilin University：Medicine Edition
基金
吉林省卫计委基金项目资助课题(20165069
2015Z009)
吉林省教育厅科学技术项目资助课题(JJKH20170828KJ)
吉林省科技厅科技发展计划项目资助课题(20180101305JC)
关键词
低剂量辐射
睾丸
内质网应激
肌醇需求酶1α
X盒结合蛋白1
low dose radiation
testis
endoplasmic reticulum stress
inositol-requiring enzyme 1α
X-box binding protein 1
