促红细胞生成素后处理对大鼠再灌注损伤肺细胞凋亡的干预及机制

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (4): 371-376.

促红细胞生成素后处理对大鼠再灌注损伤肺细胞凋亡的干预及机制

金晓盛¹, 吴厉锋¹, 钱小英¹, 邵美琴¹, 王万铁²

¹温州市人民医院呼吸内科,温州 325000,浙江;
²温州医科大学缺血再灌注损伤研究所,温州 325035,浙江

收稿日期:2013-11-06 修回日期:2014-04-17 出版日期:2014-04-26 发布日期:2020-07-24
通讯作者: 王万铁,通信作者,男,教授,主任医师,博士生导师,研究方向:脏器缺血/再灌注损伤。Tel:13587688106 E-mail:wwt@wzmc.edu.cn
作者简介:金晓盛,女,在职研究生,主治医师,研究方向:肺缺血/再灌注损伤。Tel:13777783561 E-mail:jxs19791109@163.com
基金资助:
温州市科技计划项目(Y20120001);温州市医药卫生科研项目(2013A01)

The interventive effects and mechanisms of erythropoietin postconditioning on celluar apoptosis of lung ischemia/reperfusion injury in rats

JIN Xiao-sheng¹, WU Li-feng¹, QIAN Xiao-ying¹, SHAO Mei-qin¹, WANG Wan-tie²

¹Department of Respiratory Medicine, Wenzhou People's Hospital, Wenzhou 325000, Zhejiang, China;
²Institute of Ischemia/Reperfusion Injury of Wenzhou Medical College, Wenzhou 325035, Zhejiang, China

Received:2013-11-06 Revised:2014-04-17 Online:2014-04-26 Published:2020-07-24

摘要/Abstract

摘要： 目的: 探讨促红细胞生成素后处理对再灌注损伤肺细胞凋亡的干预作用及其机制。方法: 健康雄性成年SD大鼠40只,随机分为假手术对照组(C组)、缺血/再灌注组(I/R组)、促红细胞生成素+缺血/再灌注组(EPO组)、促红细胞生成素+溶剂对照组1(0.4%DMSO的PBS溶液)(D组)和促红细胞生成素+U0126(U组)。对比观察各组肺组织湿/干重比值(W/D)的变化;原位缺口末端标记法(TUNEL)检测肺组织细胞凋亡情况,计算凋亡指数(AI);免疫组织化学方法测定肺组织Bcl-2、Bax蛋白的相对含量,RT-PCR法检测肺组织Bcl-2、Bax mRNA表达;光镜下观察肺组织的病理变化及测定肺泡损伤数(IQA)。结果: 与C组比较,I/R组肺组织W/D显著升高,I/R组IQA显著升高,AI显著升高,Bcl-2蛋白和Bcl-2 mRNA表达明显下降,Bax蛋白和Bax mRNA表达明显上调,Bcl-2/Bax的比值降低(P均<0.01),肺组织形态学发生异常改变;与I/R组比较,EPO组、D组、U组的W/D显著降低,IQA显著降低,AI显著降低,Bcl-2蛋白和Bcl-2 mRNA表达增强,Bax蛋白和Bax mRNA表达减弱,Bcl-2/Bax的比值增高(P<0.05 或P<0.01),肺组织形态学结构异常改变有所减轻;D组的W/D、IQA、AI、Bcl-2蛋白和Bcl-2 mRNA、Bax蛋白和Bax mRNA及Bcl-2/Bax的比值与EPO组变化相近(P>0.05);与EPO组比较,U组的W/D升高,IQA升高,AI显著升高,Bcl-2蛋白和Bcl-2 mRNA表达下降,Bax蛋白和Bax mRNA表达上调,Bcl-2/Bax的比值降低(P <0.05 或P<0.01),U组的肺组织形态学结构损伤较EPO组加重。结论: 促红细胞生成素后处理能减轻肺缺血/再灌注损伤,其机制可能通过激活ERK1/2信号转导通路,上调凋亡抑制因子Bcl-2的表达,下调促凋亡基因Bax的表达,提高Bcl-2/Bax比值,使肺组织细胞凋亡减少。

关键词: 促红细胞生成素, 后处理, 再灌注损伤, 肺, 细胞凋亡

Abstract: AIM: To investigate the interventive effects and mechanisms of erythropoietin postconditioning on celluar apoptosis of lung ischemia/reperfusion injury in rats. METHODS: Adult male Sprague-Dawley rats were randomly divided into 5 groups based upon the intervention (n=8): control group (group C), lung ischemia reperfusion group (group I/R), I/R+EPO(group EPO), EPO+solvent control 1(0.4%DMSO of the PBS solution)(group D), EPO+U0126(group U). At the end of the experiment, the wet to dry wight ratio(W/D) of the lung tissue was tested; the pneumocyte apoptosis index(AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end abeling(TUNEL) ; the content of Bcl-2 and Bax in cytoplasm was determined with immunohistochemistry, Bcl-2 mRNA and Bax mRNA in lung tissues were determined by RT-PCR. The pathological changes of lung tissue were observed under light microscope and the index of quantitative assessment(IQA) of histologic lung injury was counted by HE staining. RESULTS: Compared with group C, W/D, IQA and AI of lung tissue were increased in group I/R, the expression of Bcl-2 protein and Bcl-2 mRNA were decreased, the expression of Bax protein and Bax mRNA were increased, the ratio of Bcl-2/Bax was decreased.There were abnormal morphological changes under the light microscope; compared with group I/R, W/D, IQA and AI of lung were decreased in EPO, D and U groups, the expression of Bcl-2 protein and Bcl-2 mRNA were increased but the Bax protein and Bax mRNA were decreased, the ratio of Bcl-2/Bax was increased. The morphological changes under the light microscope markedly reversed in EPO, D and U groups;there was no statistical difference between group D and group EPO; compared with group EPO, W/D, IQA and AI of lung were increased in group U, the expression of Bcl-2 protein and Bcl-2 mRNA were decreased, the expression of Bax protein and Bax mRNA were increased, the ratio of Bcl-2/Bax was decreased. There were abnormal morphological changes under the light microscope in group U than group EPO. CONCLUSION: Erythropoietin postconditioning may attenuate pneumocyte apoptosis in LIRI by activating ERK1/2 signal transduction pathway, and up-regulating expression of Bcl-2/ Bax ratio.

Key words: erythropoietin, postconditioning, reperfusion injury, lung, apoptosis

中图分类号:

R965.2

金晓盛, 吴厉锋, 钱小英, 邵美琴, 王万铁. 促红细胞生成素后处理对大鼠再灌注损伤肺细胞凋亡的干预及机制[J]. 中国临床药理学与治疗学, 2014, 19(4): 371-376.

JIN Xiao-sheng, WU Li-feng, QIAN Xiao-ying, SHAO Mei-qin, WANG Wan-tie. The interventive effects and mechanisms of erythropoietin postconditioning on celluar apoptosis of lung ischemia/reperfusion injury in rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(4): 371-376.

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