一氧化碳气体干燥保存法对小鼠心脏保存效果的实验研究  

Effect of carbon monoxide preservation solution on heterotopic transplantation of isolated heart in mice

作　　者：黄晶晶[1,2] 赵明一[3] 郑少忆[1] 庄建[1] 陈寄梅[1] 黄帅[1,4] 富旗 朱小兰[1] 朱平[1] 

机构地区：[1]广东省心血管病研究所广东省人民医院广东省医学科学院,广州510010 [2]汕头大学医学院,广东汕头515041 [3]中南大学湘雅三医院,长沙410006 [4]南方医科大学,广州510282

出　　处：《岭南心血管病杂志》2014年第3期372-376,382,共6页South China Journal of Cardiovascular Diseases

基　　金：中国科技部国际合作项目(项目编号:2008DFA31140;2010DFA32660);中国科技支撑计划项目(项目编号:2011BAI11B22);国家自然科学基金项目(项目编号:81370230);人力资源与社会保障部2013年度留学人员重点项目(朱平)

摘　　要：目的观察一氧化碳（carbon monoxide，CO）气体干燥保存法对同种小鼠心脏异位移植的影响。方法采用C57BL／6雄性小鼠，建立同系小鼠心脏腹部异位移植模型。4—6周龄小鼠60只，分为3组（n=20），即空白对照组，HTK供体心组及CO供体心组；8～10周龄小鼠40只，分为HTK受体组及CO受体组（n=20）。取HTK供体心组小鼠心脏浸泡于HTK液中；取CO组小鼠心脏悬挂并置于高压气体罐中（PO2：3．2×101．3kPa＋PCO：0．8×101．3kPa=4×101．3kPa）。两组供体心均置于4℃冰箱中保存8h后，分别移植于两受体组小鼠腹部。空白对照组仅取供体心，不做移植术。移植完成后，比较HTK组及CO组复跳时间。移植2h后，对比观察两组移植心脏的颜色及复跳情况。取3组供体心心肌组织，分别采用苏木素伊红染色和TUNEL染色法对比供体心病理学改变及心肌细胞的凋亡情况。结果HTK供体心组6只复跳，CO供体心组13只复跳，复跳率差异有统计学意义[86．7％（13／20）vs.53.3％（7／20），P=0．027]；与HTK供体心组相比，CO供体心组供体心复跳时间显著缩短，差异有统计学意义[（203．92±73．52）s vs．（352．67±61．19）s，P=0．001]；病理方面，与空白对照组比较，CO供体心组供体心病理损伤明显，但与HTK组比较明显减轻；Tunel检测显示，CO组凋亡率较空白对照组升高，差异有统计学意义（P〈0．05）；但比HTK组明显减少，差异有统计学意义（P〈0．05）。结论CO气体保存法可以缩短供体小鼠心脏复跳时间及减少心肌细胞的凋亡，对离体心脏具有显著的保护作用。Objectives To evaluate the effects of heterotopic cardiac transplantation in mice with carbon monoxide （CO） preservation solution. Methods Cardiac heterotopic transplantation model was established with C57BL/6 male mice. Sixty 4 to 6-week-old age-matched male mice were divided into three donor groups （n=20） ： naive group, HTK donor group and CO donor group. Forty 8 to 10-week-old age-matched male mice were divided into two receptor groups （n= 20） ： HKT receptor group and CO receptor group. In HTK donor group, the donor hearts were extracted and preserved in 4℃ HTK solution for 8 h; in CO donor group, the donor hearts were extracted and exposed to a gas mixture （PO2： 3.2× 101.3 kPa ＋ PCO： 0.8×101.3 kPa =4×101.3 kPa） befor being preserved in a refrigerator at 4℃ for 8 h. Heterotopicheart transplantation was later performed in the abdomens of recipient mice. The donor hearts of naive group were extracted without transplantation. Durations of cardiac rebeating in the two groupos were compared. Histopathological changes of heart tissues of the 3 groups were evaluated by routine pathological examinations and myocardial apoptosis was detected by Tunnel method 2 h after transplantation. Results Six of the HTK donor cases showed resuscitation, while 13 in CO donor group. The resuscitation rates between the 2 groups were significantly different [86.7% （13/20） vs. 53.3% （7/20）, P=0.027]. Compared with HTK donor group, the resuscitation time was significantly shorter [ （203.92±73.52） s vs. （352.67±61.19） s, P=0.001 ], myocardial pathological changes were obviously less and apoptotic cell number significantly decreased in CO donor group （P〈0.05）. While being compared with naive group, the results were opposite （P〈0.05）. Conclusions CO preservation can decrease resuscitation time and reduce apoptotic cell number, which indicates that it is an effective way for myocardial preservation.

关 键 词：心脏异位移植 一氧化碳 HTK缓冲液 复跳时间 

分 类 号：R654.2[医药卫生—外科学]