OSI-027抑制mTOR信号通路减轻高氧诱导的幼鼠肺损伤  

OSI-027 alleviates hyperoxia-induced lung injury in juvenile rats by inhibiting the mTOR signaling pathway

作　　者：梁木林 刘成军[1] 党红星[1] Liang Mulin;Liu Chengjun;Dang Hongxing(Department of PICU,Children’s Hospital of Chongqing Medical University,Ministry of Education Key Laboratory of Child Development and Disorders,China International Science and Technology Cooperation Base of Child Development and Critical Disorders,Chongqing Key Laboratory Pediatrics)

机构地区：[1]重庆医科大学附属儿童医院重症医学科儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室

出　　处：《重庆医科大学学报》2019年第5期592-600,共9页Journal of Chongqing Medical University

基　　金：重庆市基础与前沿研究计划资助项目(编号:渝中科[2015]16号-13)

摘　　要：目的:探讨哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)1/2 双重抑制剂 OSI-027 对高体积分数氧(高氧)致 sprague-dawley(SD)幼鼠肺损伤及纤维化的抑制作用。方法:72 只 3 周龄 SD 幼鼠随机分为空气+生理盐水、高氧+生理盐水、高氧+雷帕霉素和高氧+OSI-027 组,分别建立动物模型(各组 n=18)。高氧干预采用 90%氧气持续处理,生理盐水、雷帕霉素、OSI-027 干预分别于观察期第 1、3、6、8、10、13 天经腹腔注射给药,在造模第 3、7、14 天取各组幼鼠测量体质量变化、肺湿干重比(wet/dry ratio,W/D)、肺组织病理学检查、肺损伤评分、肺泡间隔厚度测定、肺组织免疫组化和蛋白印迹检测 mTOR 及磷酸化核糖体 S6 蛋白激酶(pS6K1)蛋白在肺组织的分布和表达。结果:从时间因素看,各组幼鼠体质量(F 时间=297.098,P=0.000)、mTOR 免疫组化(F 时间=379.978,P=0.000)、mTOR(F 时间=166.991,P=0.000)和 pS6K1(F 时间=122.676,P=0.000)蛋白水平都随时间延长而增加。除空气组外,其余各组肺损伤评分(F 时间=1410.362,P=0.000)、肺泡间隔厚度(F 时间=356.312,P=0.000)、pS6K1 免疫组化(F 时间=57.992,P=0.000)都随时间延长而升高,肺 W/D(F 时间=28.915,P=0.000)第 3、7 天时升高,第 14 天时下降。从分组因素看,体质量(F 分组=176.597,P=0.000)空气组明显高于其他组,肺 W/D(F 分组=28.484,P=0.000)和肺泡间隔厚度(F 分组=296.223, P=0.000)空气组明显低于其他组,除第 3 天外,mTOR 免疫组化(F 分组=134.100,P=0.000)高氧组明显高于其他组,PS6K1 免疫组化(F 分组=234.697,P=0.000)、mTOR(F 分组=59.377,P=0.000)和 PS6K1(F 分组=101.837,P=0.000)蛋白印迹高氧组明显高于其他组,肺损伤评分(F 分组=2 420.076,P=0.000)高氧雷帕组明显高于其他组,高氧 OSI 组明显低于高氧组和高氧雷帕组。结论:高浓度氧可激活肺组织 mTOR 信号途径;mTOR 可能促进了高氧肺损伤纤维化的发生发展,其调控机制可能�Objective:To investigate the inhibitory effect of the mammalian target of rapamycin(mTOR) 1/2 dual inhibitor OSI-027 on hyperoxia-induced lung injury and fibrosis in juvenile Sprague-Dawley (SD) rats. Methods:A total of 72 juvenile SD rats aged 3 weeks were randomly divided into air+normal saline group,hyperoxia+normal saline group,hyperoxia+rapamycin group,and hyperoxia+ OSI-027 group,with 18 rats in each group. An animal model was established. Hyperoxia intervention was performed with 90% oxy gen,and normal saline,rapamycin,and OSI-027 interventions were performed via intraperitoneal injection on days 1,3,6,8,10,and 13 of observation,respectively. On days 3,7,and 14,the change in body weight,lung wet/dry(W/D) ratio,lung injury scores,and alve olar septal thickness were measured;lung histopathological examination was performed;immunohistochemistry and Western blot were used to evaluate the distribution and expression of mTOR and phosphorylated ribosomal S6 kinase(pS6K1) in lung tissue. Results:As for the factor of time,there were significant increases over time in body weight (Ftime=297.098,P=0.000), immunohistochemistry of mTOR(Ftime=379.978,P=0.000),mTOR(Ftime=166.991,P=0.000),and pS6K1(Ftime=122.676,P=0.000). All groups except the air+normal saline group had significant increases in lung injury scores(Ftime=1 410.362,P=0.000),alveolar septum thickness(Ftime=356.312,P=0.000),and pS6K1 immunohistochemistry(Ftime=57.992,P=0.000) over time,as well as an increase in lung W/D ratio on days 3 and 7(Ftime=28.915,P=0.000) and a reduction in lung W/D ratio on day 14. As for the factor of grouping,the air+normal saline group had a significantly higher body weight(Fgroup= 176.597,P=0.000) and significantly lower lung W/D ratio(Fgroup=28.484,P=0.000) and alveolar septum thickness(Fgroup=296.223,P= 0.000) than the other groups. At all time points except day 3,the hyperoxia+normal saline group had significantly higher mTOR immunohistochemistry(Fgroup=134.100,P=0.000),pS6K1 immunohistochemistry(Fgroup=234.697,P=0.000),mTOR(F

关 键 词：高体积分数氧 肺损伤纤维化 哺乳动物雷帕霉素靶蛋白 雷帕霉素 OSI-027 

分 类 号：R725.6[医药卫生—儿科]