高压干燥保存对小鼠供心冷缺血再灌注损伤中自噬及凋亡作用的影响  

Autophagy and cardiocyte apoptosis after heterotopic transplantation of the mouse heart preserved in highpressured mixed gas

作　　者：张瑞[1] 黄帅[2] 富旗 郑少忆[2] 郭惠明[2] 陈寄梅[2] 庄建[2] 朱平[1] 

机构地区：[1]南方医科大学研究生学院,广州医学硕士研究生510515 [2]广东省人民医院心外科,广州510080

出　　处：《医学研究生学报》2015年第12期1236-1241,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81370230);国家科技部国际科技合作项目(2010DFA32660)

摘　　要：目的心脏移植是治疗终末期心脏病的有效手段,延长供心保存时间有助于解决供心紧缺的问题,文中旨在通过观察一氧化碳和氧气的高压干燥保存是否能够延长供心保存时间及其作用机制。方法选取SPF级C57BL/6雄性小鼠共84只,将4～6周龄48只供鼠随机分成4组（n=12）：对照组（仅取供体心,不做移植）、即刻移植供体组（供心不经保存取下后即刻移植）、组氨酸-色氨酸-酮戊二酸盐液（histidine-tryptophan-ketoglutarate solution,HTK）保存供体组（供心浸泡于HTK液中24 h）及高压干燥保存供体组[供心悬挂并置于高压气体罐中（氧分压：3200 h Pa,一氧化碳分压：800 h Pa）,保存24 h]。将6～8周龄36只受鼠随机分为3组（n=12）：即刻移植受体组、HTK保存受体组及高压干燥保存受体组。将各供体组的供心分别对应移植于各受体组,移植后2 h,对比观察各受体组复跳情况及供心功能。取对照组及各移植受体组供心组织,HE染色观察供心病理学改变,Tunnel染色观察心肌细胞凋亡情况,Western blot检测自噬标志性蛋白LC3-Ⅱ及Bcl-2蛋白表达情况。结果即刻移植受体组与高压干燥保存受体组复跳率高于HTK保存受体组（P〈0.05）。高压干燥保存受体组、HTK保存受体组、即刻移植受体组移植后2 h供心功能评分分别为[2.5（2.0～2.9）、0.8（0.5～1.0）、4.5（4.0～4.5）分],组间两两比较差异均有统计学意义（P〈0.05）。高压干燥保存受体组LC3-Ⅱ蛋白及Bcl-2蛋白表达量为（2.06±0.29、0.87±0.18）、即刻移植受体组为（1.24±0.20、2.07±0.32）,均高于对照组（0.13±0.03、0.19±0.02）与HTK保存受体组（0.16±0.06、0.26±0.08）,差异有统计学意义（P〈0.05）。HTK保存受体组Bcl-2蛋白表达量高于对照组（P〈0.05）。高压干燥保存受体组LC3-Ⅱ蛋白低于即刻移植受体组（P〈0.05）。HE染色结果显示：高压干燥保存受体组供心心�Objective Heart transplantation is an effective treatment of end-stage heart diseases and extending the time of donor heart preservation helps to make up for the shortage of donor hearts.This study was to investigate whether high-pressured mixed gas（ HPMG） of carbon monoxide and oxygen could prolong the time of donor heart preservation and its mechanisms. Methods Forty-eight C57 BL /6 male mice aged 4- 6 weeks were randomly divided into four groups of equal number： control（ the donor heart isolated but not transplanted）,immediate transplantation（ the donor heart transplanted right after isolated）,HTK-preservation（ the donor heart preserved in histidine-tryptophan-ketoglutarate solution for 24 hours after isolated,and HPMG preservation（ the donor heart preserved in an HPMG chamber with the oxygen partial pressure of 3200 h Pa and carbon monoxide partial pressure of 800 h Pa for 24 hours after isolated）. Another 36 recipient mice aged 6- 8 weeks were randomly assigned to receive the donor heart immediately after harvested（ n = 12）,preserved in HTK solution（ n = 12）,or preserved in HPMG（ n = 12）. At 2 hours after transplantation,the status of heart re-beating and cardiac function were compared among different groups of recipient mice. At 24 hours,tissues were taken from the transplanted hearts for examination of pathologic changes by HE staining,detection of the apoptosis of cardiac cells by TUNEL,and determination of the expressions of microtubule-associated protein 1light chain 3-Ⅱ（ LC3-Ⅱ） and B cell lymphoma/leukemia-2（ Bcl-2） by Western blot. Results The re-beating rates of the immediately transplanted and HPMG-preserved hearts were significantly higher than that of the HTK-preserved ones（ P〈0. 05）. At 2 hours after transplantation,the cardiac function scores were 2. 5（ 2. 0- 2. 9）,0. 8（ 0. 5- 1. 0）,and 4. 5（ 4. 0- 4. 5） in the immediate implantation,HPMG-preservation and HTK-preservation groups respectively,with statistically significant differences

关 键 词：高压干燥 一氧化碳 离体供心保存 冷缺血再灌注损伤 心肌微管相关蛋白1-轻链3-Ⅱ B细胞白血病/淋巴瘤-2 自噬 凋亡 

分 类 号：R654.2[医药卫生—外科学]