摘要
目的探讨心肌缺血再灌注致肠损伤的机制并评价缺氧诱导因子-1α(HIF-1α)对心肌缺血再灌注诱导肠损伤的影响。方法将40只18~20月龄健康清洁级C57BL/6雄性小鼠(体重24~30 g)按照随机数字表法分为假手术组(S组)、心肌缺血再灌注组(IR组)、心肌缺血再灌注+HIF-1α激动剂DMOG组(IRD组)、心肌缺血再灌注+HIF-1α抑制剂2ME2组(IRM组),每组10只。除S组外,其余三组结扎冠状动脉左前降支,缺血30 min再灌注2 h,建立心肌缺血再灌注模型;以术中小鼠左心室变白、心电图ST段抬高为结扎成功标志。S组仅穿线不结扎。IRD组术前24 h腹腔注射40 mg/kg的DMOG;IRM组术前30 min腹腔注射15 mg/kg的2ME2;其余组注射同等体积生理盐水。检测小鼠左室射血分数(LVEF)、左室短轴缩短率(LVFS);观察肠组织病理学改变并采用Chiu’s评分定量;比较各组肠型脂肪酸结合蛋白(I-FABP)、二胺氧化酶(DAO)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量;比较各组肠组织HIF-1α、NOD样受体热蛋白结构域相关蛋白3(NLRP3)及Occludin表达。结果与S组比较,IR组LVEF、LVFS降低(P<0.05)。S组肠上皮细胞完整,层次清晰;IR组肠道组织完整性被破坏,小肠不规则排列的微绒毛部分脱落,间质内较多炎症细胞浸润;IRD组肠道组织较完整,部分肠绒毛变钝;IRM组肠道完整性被破坏,出现细胞脱落现象,炎症浸润明显。与S组比较,IR组Chiu’s评分升高(P<0.05)。与IR组比较,IRD组Chiu’s评分降低,IRM组Chiu’s评分升高(P<0.05)。与S组比较,IR组HIF-1α、NLRP3表达及I-FABP、DAO、TNF-α和IL-1β含量升高,Occludin表达降低(P<0.05)。与IR组比较,IRD组HIF-1α、Occludin表达升高,NLRP3表达及I-FABP、DAO、TNF-α和IL-1β含量降低(P<0.05);IRM组HIF-1α、Occludin表达降低,NLRP3表达及I-FABP、DAO和IL-1β含量升高(P<0.05)。结论心肌缺血再灌注可诱导肠道炎症反应发生;肠组织稳定HIF-1α表达可减轻心肌缺血再灌注致肠损伤,其机制可能与促进肠道屏障结构完整性、抑制NLRP3介导的炎症反应有关。
Objective To explore the mechanism of intestinal injury caused by myocardial ischemia reperfusion and to evaluate the effect of hypoxia-induced factor-1α(HIF-1α)on intestinal injury induced by myocardial ischemia reperfusion.Methods A total of 40 healthy 18-20 months old C57BL/6 males(weight 24-30 g)were divided into sham group(S group),myocardial ischemia reperfusion group(IR group),myocardial ischemia reperfusion+HIF-1αagonist DMOG group(IRD group),myocardial ischemia reperfusion+HIF-1αinhibitor 2ME2 group(IRM group),with 10 mice in each group.Except for S group,the other three groups underwent ligation of left anterior descending coronary artery ischemia for 2 h to establish a myocardial ischemia reperfusion model.During the operation,the left ventricular whitening and ST segment elevation of electrocardiogram were markers of successful ligation.S group was only threaded without ligature.In IRD group,40 mg/kg DMOG was injected 24 h before surgery;in IRM group,15 mg/kg 2ME2 was injected 30 min before surgery;the other groups were injected with equivalent volume of saline.Left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)were detected;intestinal histopathological changes were observed and quantified by Chiu’s score;the contents of intestinal fatty acid-bounding protein(I-FABP),diamine oxidase(DAO),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)of all groups were compared;and the expression of intestinal HIF-1α,NOD-like receptor thermal protein domain associated protein 3(NLRP3),and Occludin of all groups were compared.Results Compared with S group,LVEF and LVFS in IR group were decreased(P<0.05).The intestinal epithelial cells in S group were intact and the layers were clear;in IR group,the integrity of the intestinal tissue was compromised,with some irregularly arranged microvilli in the small intestine being partially shed,and a significant infiltration of inflammatory cells in the stroma;in IRD group,the intestinal tissue was relatively intact,with some villi becoming blunted;in IRM group,the intestinal integrity was compromised,with cells shedding and a marked infiltration of inflammation.Compared with S group,the Chiu’s scores of IR group were increased(P<0.05).Compared with IR group,the Chiu’s scores of of IRD group were decreased(P<0.05);the Chiu’s scores of IRM group were increased(P<0.05).Compared with S group,the expressions of HIF-1αand NLRP3 and the contents of I-FABP,DAO,TNF-α,and IL-1βin IR group were increased,while the expression of Occludin were decreased(P<0.05).Compared with IR group,the expressions of HIF-1αand Occludin expression were increased,while the expression of NLRP3 and contents of I-FABP,DAO,TNF-α,and IL-1βwere decreased in IRD group(P<0.05);the expressions of HIF-1αand Occludin were decreased,while the expression of NLRP3 and the contents of I-FABP,DAO,and IL-1βwere increased in IRM group(P<0.05).Conclusion Myocardial ischemia reperfusion can induce intestinal inflammatory response;stable HIF-1αexpression can reduce myocardial ischemia reperfusion,the mechanism is related to promote the structural integrity of intestinal barrier and inhibit NLRP3-mediated inflammatory response.
作者
朱阔
程虎
于文彬
杨雪
坎安·吐尔逊
王江
ZHU Kuo;CHENG Hu;YU Wenbin;YANG Xue;Kanan Tuerxun;WANG Jiang(Department of Anesthesiology,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi830000,China)
出处
《中国医药导报》
2025年第5期8-12,共5页
China Medical Herald
基金
国家自然科学基金地区科学基金项目(81960053)。
关键词
缺氧诱导因子-1Α
心肌缺血再灌注
肠损伤
紧密连接蛋白
炎症
Hypoxia-inducible factor-1α
Myocardial ischemia reperfusion
Intestinal injury
Tight junction proteins
Inflammation
作者简介
朱阔(1998-),男,新疆医科大学第一临床医学院2022级麻醉学专业在读硕士研究生,研究方向:心肌缺血再灌注致肠损伤;通讯作者:王江(1967-),女,博士,主任医师,研究方向:体外循环与心肌保护。
