低剂量辐射诱导小鼠睾丸细胞中IRE1α和XBP1 mRNA及其蛋白的表达

doi:10.13481/j.1671-587x.20180506

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (05): 919-923.doi: 10.13481/j.1671-587x.20180506

低剂量辐射诱导小鼠睾丸细胞中IRE1α和XBP1 mRNA及其蛋白的表达

于雷¹, 唐庚², 方芳², 李鑫², 龚守良², 王志成², 王剑锋³

1. 吉林大学第二医院放疗科, 吉林长春 130041;
2. 吉林大学公共卫生学院卫生部放射生物学重点实验室, 吉林长春 130021;
3. 吉林大学中日联谊医院放疗科, 吉林长春 130033

收稿日期:2017-10-20 出版日期:2018-09-28 发布日期:2018-11-20
通讯作者: 王剑锋,主治医师,医学博士(Tel:0431-84995413,E-mail:radiation1978@163.com);王志成,医学博士,副教授,硕士研究生导师(Tel:0431-85619343,E-mail:zhicheng@jlu.edu.cn) E-mail:radiation1978@163.com;zhicheng@jlu.edu.cn
作者简介:于雷(1977-),男,吉林省长春市人,副主任医师,医学博士,主要从事辐射肿瘤学方面的研究。
基金资助:
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）

Expressions of IRE1α and XBP1 mRNA and proteins in mouse testis cells induced by low dose ionizing radiation

YU Lei¹, TANG Geng², FANG Fang², LI Xin², GONG Shouliang², WANG Zhicheng², WANG Jianfeng³

1. Department of Radiotherapy, Second Hospital, Jilin University, Changchun 130041, China;
2. Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China;
3. Department of Radiotherapy, China-Japan Union Hospital, Jilin University, Changchun 130033, China

Received:2017-10-20 Online:2018-09-28 Published:2018-11-20

摘要/Abstract

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200 mGy）X射线照射后12 h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：75 mGy X射线照射后0~24 h，小鼠睾丸细胞中IRE1α、T-XBP1和S-XBP1 mRNA（除了3 h时T-XBP1和S-XBP1 mRNA）表达水平均随时间延长而升高，并在6 h时达到峰值，而后逐渐降低；与0 h组比较，6、12和24 h组IRE1α mRNA、6和12 h组T-XBP1 mRNA及S-XBP1 mRNA表达水平均明显升高（P<0.05或P<0.01）。与0 h组比较，不同时间组IRE1α和S-XBP1蛋白表达强度升高，6和12 h组IRE1α蛋白表达强度升高最明显，24 h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。0~200 mGy照射后12 h，小鼠睾丸细胞中IRE1α、T-XBP1和S-XBP1 mRNA表达水平升高，75 mGyX射线照射后升高达峰值后逐渐降低，甚至降至0 mGy以下；与0 mGy组比较，75和200 mGy组小鼠睾丸细胞中IRE1α、75 mGy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0 mGy组比较，75 mGy组IRE1α和S-XBP1蛋白表达强度升高，75和200 mGy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-XBP1 mRNA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。

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

Abstract: 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 IRE1α mRNA(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 IRE1α and 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 IRE1α mRNA in the testis cells of the mice in 75 and 200 mGy groups and the expression levels of T-XBP1 mRNA and S-XBP1 mRNA in 75 mGy group were significantly increased (P<0.05 or P<0.01); the expression intensities of IRE1α protein in 75 mGy group and S-XBP1 protein in 75 and 200 mGy groups were increased mostly,while the expression intensity of T-XBP1 protein had no obvious change. Conclusion:LDR can induce the increase of IRE1α and S-XBP1 mRNAs and proteins,and activate the IRE1-XBP1 pathway in endoplasmic reticulum stress.

Key words: low dose radiation, testis, endoplasmic reticulum stress, inositol-requiring enzyme 1α, X-box binding protein 1

中图分类号:

Q345

于雷, 唐庚, 方芳, 李鑫, 龚守良, 王志成, 王剑锋. 低剂量辐射诱导小鼠睾丸细胞中IRE1α和XBP1 mRNA及其蛋白的表达[J]. 吉林大学学报(医学版), 2018, 44(05): 919-923.

YU Lei, TANG Geng, FANG Fang, LI Xin, GONG Shouliang, WANG Zhicheng, WANG Jianfeng. Expressions of IRE1α and XBP1 mRNA and proteins in mouse testis cells induced by low dose ionizing radiation[J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 919-923.

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低剂量辐射诱导小鼠睾丸细胞中IRE1α和XBP1 mRNA及其蛋白的表达

Expressions of IRE1α and XBP1 mRNA and proteins in mouse testis cells induced by low dose ionizing radiation

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