内皮祖细胞微囊泡通过p38丝裂原活化蛋白激酶/核因子-κB通路保护心肌缺血再灌注损伤实验研究OA
Experimental study of endothelial progenitor cell-derived microvesicles protect myocardial ischemia-reperfusion injury through the p38 MAPK/NF-κB pathway
目的:探究内皮祖细胞微囊泡(EPC-MVs)对心肌缺血再灌注损伤(MIRI)的保护作用.方法:采用密度梯度离心法以及差数贴壁法分离和培养内皮祖细胞(EPCs),收集EPCs并离心获取微囊泡(MVs).大鼠分为假手术组、MIRI组、EPC-MVs组,每组3只.采用超声心动图检测大鼠心功能;ELISA检测心肌乳酸脱氢酶(LDH)、肌酸激酶同工酶-MB(CK-MB)、心肌肌钙蛋白I(cTnI)水平及肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平;HE染色观察心肌组织病理情况;Western blot检测心肌组织p38丝裂原活化蛋白激酶(p38 MAPK)和核因子-κB(NF-κB)表达水平.结果:EPC-MVs组较MIRI组左心室收缩末期内径(LVESD)、左心室舒张末期内径(LVEDD)显著降低,左心室射血分数(EF)、短轴缩短分数(FS)显著升高(均P＜0.05);EPC-MVs组大鼠血清LDH、CK-MB、cTnI水平显著降低(均P＜0.05).EPC-MVs组大鼠心肌纤维排列较整齐,肌纤维断裂较MIRI组减少,细胞间有较少的炎性细胞浸润.MIRI组大鼠炎症因子水平(IL-1β、TNF-α)、p-p38 MAPK、p-NF-κB水平显著升高(均P＜0.01);与MIRI组比较,EPC-MVs组大鼠炎症因子水平、p-p38 MAPK、p-NF-κB水平显著降低(均P＜0.01).结论:EPC-MVs对MIRI大鼠具有保护作用,其机制可能通过影响p38 MAPK/NF-κB信号通路进而影响炎症反应.
Objective:To investigate the cardioprotective effects of endothelial progenitor cell-derived microvesi-cles(EPC-MVs)against myocardial ischemia-reperfusion injury(MIRI).Methods:Endothelial progenitor cells(EPCs)were isolated and cultured using density gradient centrifugation combined with differential adhesion meth-ods.EPC-MVs were collected via ultracentrifugation.Sprague-Dawley rats were randomized into three groups(n=3 per group):sham-operated,MIRI,and EPC-MVs-treated.Cardiac function was assessed by transthoracic echocardio-graphy to measure left ventricular end-systolic diameter(LVESD),end-diastolic diameter(LVEDD),ejection frac-tion(EF),and fractional shortening(FS).Serum levels of lactate dehydrogenase(LDH),creatine kinase-MB(CK-MB),cardiac troponin I(cTnI),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β)were quantified by enzyme-linked immunosorbent assay(ELISA).Histopathological alterations in myocardial tissue were evaluated via hema-toxylin-eosin(HE)staining.Protein expression levels of phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK)and nuclear factor-kappa B(p-NF-κB)were analyzed by Western blot.Results:Compared to the MIRI group,the EPC-MVs group exhibited significantly reduced LVESD and LVEDD(all P＜0.05),accompanied by in-creased EF and FS(all P＜0.05).Serum biomarkers of myocardial injury(LDH,CK-MB,cTnI)and pro-inflamma-tory cytokines(TNF-α,IL-1β)were markedly attenuated in the EPC-MVs group(all P＜0.05).Histopathological analysis revealed improved myocardial architecture in the EPC-MVs group,characterized by alleviated myofibril dis-array,reduced interstitial inflammatory cell infiltration,and diminished necrosis compared to the MIRI group.West-ern blot demonstrated that EPC-MVs treatment significantly suppressed MIRI-induced activation of p-p38 MAPK and p-NF-κB(all P＜0.01).Conclusion:EPC-MVs confer cardioprotection against MIRI,potentially through inhibi-tion of the p38 MAPK/NF-κB signaling pathway,thereby attenuating inflammatory cascades.
宋艳玲;陈明慧;刘皇军
长江大学附属第一医院,湖北荆州 434000海南医科大学第一附属医院,海南海口 570100长江大学附属第一医院,湖北荆州 434000
医药卫生
心肌缺血再灌注损伤内皮祖细胞微囊泡细胞通路炎症心肌保护信号转导
Myocardial ischemia-reperfusion injuryEndothelial progenitor cell microvesiclesCellular path-waysInflammationCardioprotectionSignal transduction
《陕西医学杂志》 2026 (1)
29-34,6
海南省自然科学基金资助项目(823RC577)
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