摘要
Background The maintenance of heart viability is important for heart transplantation. Currently, heart preservation is limited to 6 hours of cold ischemic storage. This study explored a new heart preservation method under a high-pressured mixed gas chamber. Methods C57BL/6 male mice were used to establish the model of mice cervical heterotopic heart transplantation. Adult donor mice were randomly divided into three groups subjected to naive operation （Group A）, standard control （Group B） and experimental control （Group C）. The recipient mice were randomly divided into two groups subjected to standard control and experimental control. Group A： hearts were isolated; Group B： hearts were isolated and preserved in HTK solution at 4 ℃ for 8 h and transplanted; Group C： hearts were isolated and preserved in high pressured gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） at 4 ℃ for 8h and transplanted. After transplantation, the state of re-beating and cardiac function were observed for Group B and C. At 24 h after transplantation, samples were collected for HE staining, cardiac cell apoptosis detection by Tunnel staining and analysis of tumor necrosis factor α （TNF-α）, interleukin-1β （IL-1β）, interleukin-6 （IL-6） and interleukin-10 （IL-10） by reverse transcriotion-polymerase chain reaction （RT-PCR）. Results In group C, 15 transplanted hearts were re-beat, while only 6 in Group B. The re-beating rate in Group C was significantly higher than Group B [75.0%（15/20） vs. 30.0%（6/20） ,P = 0.01]. The time of re-beating was significantly different between Group B, and C [（352.35 ± 61.07）s vs. （207.85 ± 71.24） s, P 〈 0.011. HE staining showed that pathologic changes such as ceil edema and inflammatory cell infiltration were more obvious in Group B and C than in Group A, but less obvious in Group C compared with Group B. Tunnel staining showed that Group B had more obvious apoptosis than Group A and C. RT-PCR results showed significant increase of TNF-α, IL-1β and IL-6 expression in Group B than Group C （P 〈 0.01, the expression of IL-10 was higher in Group C than that in Group B. Conclusion Highpressured mixed gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） preservation can reduce cold ischemia and reperfusion injury of donor heart, therefore to maintain myocardial viability and prolong preservation time of donor heart.
Background The maintenance of heart viability is important for heart transplantation. Currently, heart preservation is limited to 6 hours of cold ischemic storage. This study explored a new heart preservation method under a high-pressured mixed gas chamber. Methods C57BL/6 male mice were used to establish the model of mice cervical heterotopic heart transplantation. Adult donor mice were randomly divided into three groups subjected to naive operation （Group A）, standard control （Group B） and experimental control （Group C）. The recipient mice were randomly divided into two groups subjected to standard control and experimental control. Group A： hearts were isolated; Group B： hearts were isolated and preserved in HTK solution at 4 ℃ for 8 h and transplanted; Group C： hearts were isolated and preserved in high pressured gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） at 4 ℃ for 8h and transplanted. After transplantation, the state of re-beating and cardiac function were observed for Group B and C. At 24 h after transplantation, samples were collected for HE staining, cardiac cell apoptosis detection by Tunnel staining and analysis of tumor necrosis factor α （TNF-α）, interleukin-1β （IL-1β）, interleukin-6 （IL-6） and interleukin-10 （IL-10） by reverse transcriotion-polymerase chain reaction （RT-PCR）. Results In group C, 15 transplanted hearts were re-beat, while only 6 in Group B. The re-beating rate in Group C was significantly higher than Group B [75.0%（15/20） vs. 30.0%（6/20） ,P = 0.01]. The time of re-beating was significantly different between Group B, and C [（352.35 ± 61.07）s vs. （207.85 ± 71.24） s, P 〈 0.011. HE staining showed that pathologic changes such as ceil edema and inflammatory cell infiltration were more obvious in Group B and C than in Group A, but less obvious in Group C compared with Group B. Tunnel staining showed that Group B had more obvious apoptosis than Group A and C. RT-PCR results showed significant increase of TNF-α, IL-1β and IL-6 expression in Group B than Group C （P 〈 0.01, the expression of IL-10 was higher in Group C than that in Group B. Conclusion Highpressured mixed gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） preservation can reduce cold ischemia and reperfusion injury of donor heart, therefore to maintain myocardial viability and prolong preservation time of donor heart.
基金
supported by Major International(Regional)Joint Research Project of Ministry of Science and Technology of China(No.2010DFA32660)
