增加受者体内一氧化碳含量在减轻小鼠移植心缺血再灌注损伤中的作用和机制  

The increase of carbon monoxide in recipients ameliorates ischemia/reperfusion injury in a murine heart transplantation model

作　　者：张松林[1] 孙宗全[1] 冯剑锷[1] 吴龙[1] 于利[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心血管外科,武汉430022

出　　处：《中华器官移植杂志》2010年第3期157-161,共5页Chinese Journal of Organ Transplantation

摘　　要：目的 观察心脏移植前用二氯甲烷（MC）灌胃增加受者体内一氧化碳含量在减轻小鼠移植心缺血再灌注损伤中的作用及其机制。方法以Balb／c小鼠作为供、受者，建立颈部异位心脏移植模型。实验共分为4组，分别为MC100mg组（n=10）、MC500mg组（n=12）、橄榄油组（n=10）和正常对照组（n=5）。前3组在麻醉前3h分别用经橄榄油稀释的MC100mg／kg、MC500mg／kg及单纯橄榄油0．15ml对受者进行灌胃处理，然后行颈部异位心脏移植；正常对照组小鼠仅作麻醉处理，不进行心脏移植。分别于灌胃后0、1、3、6、12和24h剪尾采血，测定血液中CO的含量[用碳氧血红蛋白（COHb）占总血红蛋白的百分比表示]，并于相应时间点取心肌组织，检测心肌组织中CO的含量；各组移植后3和24h分别处死半数受者，检测移植后3和24h血清中肌钙蛋白I（cT—nI）、心肌组织中肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）、细胞凋亡与相关基因Bcl-2和BaxmRNA以及核因子kB（NF-kB）的表达，并观察移植心心肌的超微结构。结果与橄榄油组比较，MC100mg和MC500mg组受者血液中COHb浓度与心肌组织中CO含量明显升高，均在灌胃后3h达到峰值；MC100mg组和MC500mg组受者心脏移植后3和24h，能显著降低血清中cTnI水平（P〈0．01），并以MC500mg组降低最为明显；可明显下调促炎症基因TNFamRNA水平（P〈0．01），而抗炎症基因IL-10mRNA上调不明显（P〉0．05），同时可以显著上调抗凋亡基因Bcl-2mRNA水平（P〈0．01），并抑制促凋亡基因BaxmRNA的转录（P〈0．01）；心肌超微结构损伤明显减轻。正常对照组可见少量NF-kB表达，而橄榄油组、MC100mg组和MC5（10mg组NF-kB活性均显著增强（P〈0．01），但后三组之间NFxB活性的差异无统计学意义（P〉0．05）。结论诱导受者体内增加CO含量能通过其抗炎症和抗凋亡功能而减轻移植心缺血Objective To examine whether the increase of carbon monoxide （CO） induced by oral methylene chloride （MC） administration in recipients before heart transplantation would protect heart grafts against ischemia/reperfusion （I/R） injury associated with transplantation and to explore the possible mechanism. Methods Inbred male Balb/c mice were used as donors and recipients to establish cervical heart transplantation model. Recipients were treated with either MC （100 mg/kg or 500mg/kg, per os） （group MC 100 mg, n = 10; group MC 500 mg, n = 12） or olive oil （0. 15 ml, per os, group olive, n = 10） 3 h prior to anesthesia. Age-matched normal mice served as controls （group N, n = 5）. The serum COHb and the CO content of myocardial tissue were measured at 0, 1, 3, 6, 12, 24 h after oral MC administration. Half of recipients were killed at 3 and 24 h after transplantation for serum or cardiac graft samples. The serum cTnI levels, the mRNA levels of TNF-α, IL-10, Bcl-2, Bax, the protein levels of NF-kB and the ultrastructures of myocardium were examined. Results As compared with group olive, the serum COHb and tissue CO were increased significantly and peaked within 3 h in group MC 100 mg and group MC 500 rag. The serum cTnI levels in group MC 100 mg and group MC 500 mg were significantly decreased （P〈0. 01 ）, especially in group MC 500mg. The increase of CO in recipients of group MC100 mg and group MC 500 mg significantly inhibited the proinflammatory gene expression of TNF-α mRNA and the pro-apoptotic gene expression of Bax mRNA （P〈0. 01）, and increased the anti-apoptotic gene expression of Bcl-2 mRNA （P〈0. 01 ）, but did not increase the anti-inflammatory gene expression of IL-10 mRNA （P〈0. 05） in the heart grafts. As compared with group N, the myocardial NF-kB activation was increased significantly in group olive, group MC 100 mg and group MC 500 mg （P〈0. 01）, but there was no significant difference among the later three groups （P〈0. 05）. The myocardial ultr

关 键 词：一氧化碳 核因子KB 炎症 凋亡 缺血再灌注损伤 心脏移植 

分 类 号：R285.5[医药卫生—中药学]