RXR介导的自噬通路在大鼠肺缺血/再灌注损伤中的调控作用  

Regulation of retinoid X receptor-mediated-autophagy pathway in rat pulmonary ischemia/reperfusion injury

作　　者：项冰倩 高慧[1] CHEN Jun-hao 楼国强[1] 周卓琳 武垣伶 张晶晶[1] 许益笑[1] 王万铁[1] XIANG Bing-qian;GAO Hui;CHEN Jun-hao;LOU Guo-qiang;ZHOU Zhuo-lin;WU Yuan-ling;ZHANG Jing-jing;XU Yi-xiao;WANG Wan-tie(Institute of Ischemia/Reperfusion Injury,Wenzhou Medical University,Wenzhou 325035,China;Department of Pathology,Taizhou Hospital,Linhai 317000,China;School of Biomedical Sciences,University of Western Australia,Perth 6000,Australia)

机构地区：[1]温州医科大学缺血/再灌注损伤研究所,浙江温州325035 [2]浙江省台州医院病理科,浙江临海317000 [3]School of Biomedical Sciences,University of Western Australia

出　　处：《中国病理生理杂志》2018年第11期2054-2061,共8页Chinese Journal of Pathophysiology

基　　金：温州市公益性科技计划项目(No.2017Y0223)

摘　　要：目的:探讨维甲酸X受体(RXR)介导的细胞自噬通路对大鼠肺缺血/再灌注(I/R)损伤的作用及机制。方法:以雄性SPF级SD大鼠为研究对象,随机分成7组(n=10):正常对照(C)组、假手术(S)组、S+RXR激动剂9-顺式维甲酸(SRA)组、S+RXR抑制剂HX531(SH)组、I/R组、I/R+9-顺式维甲酸(RA)组和I/R+HX531(HX531)组。采用大鼠在体左侧肺门夹闭30 min、再灌注180 min的方法制备肺缺血/再灌注模型。C组不做任何处理;S组只开胸,不夹闭肺门,机械通气210 min;I/R组行左侧肺门夹闭30 min,再恢复灌注180 min;SRA组、SH组、RA组与HX531组在术前90 min腹腔注射9-顺式维甲酸(5 mg/kg);SH组和HX531组在术前30 min腹腔注射HX531(5 mg/kg)。再灌注结束后取左肺组织,行肺组织损伤评估(IQA);采用HE染色法观察肺组织病理学形态;免疫荧光标记法观察各组肺组织RXRα的表达情况;采用RT-PCR和Western blot分别检测各组大鼠自噬相关分子LC3、beclin 1和m TOR的mRNA和蛋白水平。结果:与C组、S组、SRA组和SH组相比,I/R组、RA组和HX531组肺IQA、LC3和beclin 1 mRNA及LC3-II和beclin 1蛋白均有明显升高,m TOR的mRNA及p-mTOR的蛋白水平明显降低,组织形态学结构亦有损伤性变化;与I/R组相比,RA组肺IQA、LC3和beclin 1 mRNA水平及LC3-II和beclin1蛋白水平均有明显下降,RXRα和m TOR的mRNA及p-mTOR的蛋白水平明显增高,肺组织结构损伤性的变化亦有明显减轻,而HX531组与I/R组比较无显著差异;与RA组相比,HX531组的肺IQA、LC3和beclin 1 mRNA水平及LC3-II和beclin 1蛋白水平均有明显升高,RXRα和m TOR的mRNA及p-mTOR蛋白水平明显降低,肺组织形态学结构的损伤性变化加重。结论:激活RXR可有效减轻大鼠肺缺血/再灌注损伤,对肺组织有一定的保护作用,具体机制可能与其抑制细胞自噬通路有关。AIM:To investigate the regulation of retinoid X receptor(RXR)-mediated autophagy pathway in rat pulmonary ischemia/reperfusion(I/R)injury.METHODS:The male SPF-grade SD rats were randomly divided into 7 groups(n=10):normal control(C)group,sham(S)group,sham plus 9-cis-retinoic acid(SRA)group,sham plus HX531(SH)group,I/R group,I/R plus 9-cis-retinoic acid(RA)group and I/R plus HX531(HX531)group.The model of pulmonary I/R injury was established by clamping the left hilum of lung for 30 min followed by 180 min of reperfusion.The animals in C group didn’t receive any treatment.Only sternotomy was performed for the rats in S group,the hilum of lung was not clamped,and the rats were mechanically ventilated for 210 min.The clamping of the left hilum of lung for 30 min followed by 180 min of reperfusion was performed in I/R group.In SRA,SH,RA and HX531 groups,9-cis-re-tinoic acid(5 mg/kg)was intraperitoneally injected at 90 min before establishment of the model.In SH group and HX531 group,HX531 at 5 mg/kg was intraperitoneally injected at 30 min before establishment of the model.Left lung tissues were removed after 180 min of reperfusion for determing the index of quantitative assessment(IQA)of alveolar damage.The pathological changes of the lung were observed by HE staining,and immunofluorescence staining was used to observe the changes of RXRαin various lung tissues.The mRNA expression of autophagy-associated molecules LC3,beclin 1 and mTOR was detected by RT-PCR.The protein levels LC3-II,beclin 1 and p-mTOR in each group were determined by Western blot.RESULTS:Compared with C group,the lung IQA,the mRNA expression of LC3 and beclin 1,and the protein levels of LC3-II and beclin 1 in I/R,RA and HX531 groups were increased significantly,the mTOR mRNA and p-mTOR protein levels were decreased(P<0.05),and the morphological structure of the lung was also impaired.Compared with I/R group,the lung IQA and the expression of LC3 and beclin 1 at mRNA and protein levels were significantly decreased,the mRNA expression of RXRαan

关 键 词：维甲酸X受体 缺血/再灌注损伤 肺 自噬 大鼠 

分 类 号：R563[医药卫生—呼吸系统] R363[医药卫生—内科学]