摘要
目的:探讨细胞自噬在大鼠缺血/再灌注肺损伤中的作用。方法:随机将40只SD大鼠分为5组(n=8),分别为①假手术组(Sham组):只开胸3.5 h;②缺血/再灌注组(I/R组):开胸夹闭肺门缺血0.5 h后再灌注3 h;③溶剂组(DMSO组):术前1 h腹腔注射DMSO溶液;④自噬激动剂组(Rap组):术前腹腔注射雷帕霉素溶液;⑤自噬抑制剂组(3-MA组):术前1 h腹腔注射3-MA溶液;后三组的其余操作同I/R组。实验结束后处死大鼠,取肺组织,记录并计算肺组织湿/干重比(W/D)、总肺含水量变化(TLW),光镜和电镜观察肺组织及细胞形态,计算肺泡损伤率(IAR),Western blot检测自噬相关蛋白的表达情况。结果:相对于sham组,其余四组肺W/D、TLW、IAR均明显升高,自噬相关蛋白表达明显上升,p-AMPK、Beclin 1、LC3 II蛋白明显增多,p-mTOR、p62蛋白明显减少(P<0.05或P<0.01),光镜下其余各组肺组织有不同程度的水肿渗出,肺泡结构紊乱,电镜下细胞超微结构损伤加重,部分可见自噬小体;与DMSO组相比,3-MA组肺W/D、TLW、IAR明显下降(P<0.05或P<0.01),自噬相关蛋白表达明显下降,肺间质水肿较轻,细胞渗出较少,细胞超微结构损伤减轻,未见自噬小体。而I/R、DMSO、Rap组的各项指标变化无统计学差异(P>0.05)。结论:肺缺血/再灌注可诱发细胞自噬增强,从而引起大鼠肺损伤。
Objective:To investigate the role of cell autophagy in lung ischemia/reperfusion injury in rats.Methods:Forty SD rats were randomly divided into 5 groups(n=8):①Sham operated group(sham group):just open rat chest for 3.5 h;②Ischemia/reperfusion group(I/R group):after open chest,clamp pulmonary hilus for 0.5h then reperfusion for 3 h;③Solvent group(DMSO group):intraperitoneal injection of DMSO solution for 1h before operation;④Autophagic inhibitor group(3-MA group);⑤Autophagic agonist group(Rap group):intraperitoneal injection of autophagic agonist rapamycin before operation;the rest operations of DMSO,3-MA and Rap groups are the same as that of I/R group.At the end of the experiment,the rats were killed by euthanasia-killing.The lung tissues were collected and the wet/dry weight ratio(W/D)and total lung water content(TLW)of the lung tissues were detected.The lung tissue structure and cell ultramicro morphology were observed by light microscopy and electron microscopy and the injuried alveolar rate(IAR)was calculated.The autophagy-related protein expressions were detected by Western blot.Results:Compared with sham group,the levels of W/D,TLW and IAR were increased,the expressions of autophagy related protein and p-AMPK,Beclin 1,LC3 II were also increased in other four groups,while the protein expressions of p-mTOR and p62 were decreased significantly(P<0.05 or P<0.01).Under the light microscope,the other groups of lung tissue had edema and exudation in varying degrees,the structure of alveoli was disordered,the ultrastructural damage of cells was aggravated under the electron microscope,and autophagosome could be observed.Compared with DMSO group,the expressions of autophagy related protein,the levels of W/D,TLW and IAR in 3-MA group were decreased(P<0.05 or P<0.01),the edema of lung interstitial was lighter,and less cells were found in alveolar cavity.Ultrastructural damage was also lighter and with less autophagosome.Besides,there was no significant difference among I/R,DMSO and Rap groups(P>0.05).Concl
作者
郝卯林
楼国强
刘秀洁
钱巍
王嘉
周卓琳
王万铁
HAO Mao-lin;LOU Guo-qiang;LIU Xiu-jie;QIAN Wei;WANG Jia;ZHOU Zhuo-lin;WANG Wan-tie(Institute of Ischemia-Reperfusion Injury,Wenzhou Medical University,Wenzhou 325035;General Medical Department,the First People's Hospital of Hangzhou Lin'an District,Hangzhou 311300;Library of Hangzhou Medical College,Hangzhou 310052,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2021年第4期385-389,共5页
Chinese Journal of Applied Physiology
基金
杭州市2018年社会发展科研入库立项补助项目(201811)
温州市基础性科研项目(Y20180093)。
