一氧化碳干燥保存方法对离体小鼠心脏的保护作用

The preservation of isolated mouse hearts by desiccation using carbon monoxide

目的：探讨一氧化碳（CO）干燥环境对离体小鼠心脏的保护作用，为心脏移植提供一种新的保存方式。方法采用C57BL／6雄性健康小鼠，建立同系小鼠心脏颈部异位移植模型。4～6周龄C57BL／6小鼠60只作为供体，随机分为3组（n＝20），即空白对照组（不做心脏移植）、HTK浸泡供体组及CO干燥供体组；8～10周龄C57BL／6小鼠40只作为受体，随机分为两组（n＝20），分别为HTK浸泡受体组、CO干燥受体组。HTK浸泡供体组及CO干燥供体组小鼠心脏分别用两种方法（HTK浸泡保存和CO高压干燥保存）在4℃环境下保存24h，通过建立同系小鼠颈部异位移植模型，监测模型复灌2h后供心复跳率，并在24h后检测供体心肌LDH、CPK、OH－1等酶学指标，观察心肌结构的改变。结果（1）复灌2h后，CO干燥受体组复跳15只，HTK浸泡受体组复跳7只，CO干燥受体组复跳率明显高于HTK浸泡受体组（P＝0.011）。（2）复灌24h后，CO干燥受体组LDH、CPK均明显低于HTK浸泡受体组（P＜0.05）；CO干燥受体组的LDH与空白对照组之间差异无统计学意义（P＝0.946），而CO干燥受体组CPK明显高于空白对照组（P＜0.001）；HTK浸泡受体组的LDH和CPK均明显高于空白对照组；而CO干燥受体组HO－1明显高于HTK浸泡受体组（P＜0.05），而HTK浸泡受体组与空白对照组之间差异无统计学意义（P＝0.449）。（3）心肌结构上，CO干燥受体组和HTK浸泡受体组较空白对照组均有明显改变，两组均可见心肌水肿及炎细胞浸润，而CO干燥受体组与HTK浸泡受体组相比，病理损伤较轻。结论与传统HTK浸泡保存相比，CO高压干燥保存方式能够减少心肌损伤，减轻炎症反应，在供心较长时间保存方面更有优势。

Objective To investigate the protective effect of desiccation using carbon monoxide on isolated mouse hearts, and to provide a new preservation method for heart transplantation .Methods The modified Heron＇s technique for heterotopic cervical cardiotransplantation was used .C57BL/6 male mice （n=100） were used in this research.The donor mice （n=60, 4-6 weeks age） were randomly divided into 3 groups, HTK group （n=20）, CO group （n=20） and Na-ive group （n=20）.The receptor mice （n=40, 8-10 weeks age） were randomly divided into 2 groups, HTK group and CO group.The donor hearts from HTK group were preserved in HTK cardioplegic solution at 4℃ for 24 hours, and then transplanted to receptors .The donor hearts from CO group were hung in a high -pressure tank （ PO2 =3 200 hPa, PCO=800 hPa） at 4℃for 24 hours, and then transplanted to receptors .Meanwhile, the hearts from Nave group received no preservation.The re beat number of the donor heart was tested .The HO-1, LDH and CPK in myocardial cell 24 hours after reperfusion of the donor heart were assessed .Results The re-beat heart counts in HTK and CO groups were 7 （35%） and 15 （75%）, with significant difference between them （P〈0.05）.The HO-1 in CO group was significantly higher than that in Naive group and HTK group .The LDH and CPK in CO group were significantly lower than those in HTK group （ P〈0.05 ） .Comparing with Naive group , pathological changes were obviously observed in CO and HTK groups, including edema and inflammatory cell invasion;however, the injury was more prominent in HTK group .Conclusion Comparing with the traditional HTK preservation , the method of preservation by desiccation of using carbon monox-ide has a better effect for mouse isolated hearts for a long time preservation .