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 u...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-展开更多
目的应用大鼠原位肝脏移植模型,研究自体吞噬(自噬)如何参与缺血再灌注损伤后肝细胞死亡过程。方法在肝脏UW保存液中4°C保存24h后行大鼠同种原位肝脏移植;渥曼青霉素治疗组(WM-G)为UW保存液中添加渥曼青霉素(100 n M),对照组(UW-G)...目的应用大鼠原位肝脏移植模型,研究自体吞噬(自噬)如何参与缺血再灌注损伤后肝细胞死亡过程。方法在肝脏UW保存液中4°C保存24h后行大鼠同种原位肝脏移植;渥曼青霉素治疗组(WM-G)为UW保存液中添加渥曼青霉素(100 n M),对照组(UW-G)为UW治疗组:大鼠原位肝移植后观察受体生存率,分别于肝移植后15、30、60及120min检测血液血清谷草转氨酶(AST)及谷丙转氨酶(ALT),肝脏标本行光学显微镜及透射电子显微镜组织形态学分析,免疫印迹方法进行自噬标记分子LC3定量。结果移植15min后,UW-G细胞质中含有大量吞噬小体和吞噬溶酶体的小团肝细胞自肝细胞索分离,分离的肝细胞堵塞肝窦内皮间隙从而引起移植2h后大面积的肝细胞坏死,坏死的肝细胞中包含非活化半胱氨酸天冬氨酸酶-3及非活化半胱氨酸天冬氨酸酶-7的调亡染色阳性核;自体吞噬标志性蛋白LC3-II的表达各组都逐渐增加,移植30min后达到高峰,WM-G可明显提高肝移植受体存活率,明显降低AST及ALT,WM-G移植15min后肝细胞中LC3-II的表达低于UW-G;超微结构显示移植15min后WM-G肝细胞中早期自体吞噬性液泡(AVi)、晚期自体吞噬性液泡(AVd)、自噬溶酶体/溶酶体(包括致密体)的比例明显低于UW-G(P<0.01)。结论自噬参与了肝脏冷缺血再灌注损伤过程,抑制自噬可降低肝移植术后冷缺血再灌注损伤。展开更多
基金supported by Major International(Regional)Joint Research Project of Ministry of Science and Technology of China(No.2010DFA32660)
文摘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-
文摘目的应用大鼠原位肝脏移植模型,研究自体吞噬(自噬)如何参与缺血再灌注损伤后肝细胞死亡过程。方法在肝脏UW保存液中4°C保存24h后行大鼠同种原位肝脏移植;渥曼青霉素治疗组(WM-G)为UW保存液中添加渥曼青霉素(100 n M),对照组(UW-G)为UW治疗组:大鼠原位肝移植后观察受体生存率,分别于肝移植后15、30、60及120min检测血液血清谷草转氨酶(AST)及谷丙转氨酶(ALT),肝脏标本行光学显微镜及透射电子显微镜组织形态学分析,免疫印迹方法进行自噬标记分子LC3定量。结果移植15min后,UW-G细胞质中含有大量吞噬小体和吞噬溶酶体的小团肝细胞自肝细胞索分离,分离的肝细胞堵塞肝窦内皮间隙从而引起移植2h后大面积的肝细胞坏死,坏死的肝细胞中包含非活化半胱氨酸天冬氨酸酶-3及非活化半胱氨酸天冬氨酸酶-7的调亡染色阳性核;自体吞噬标志性蛋白LC3-II的表达各组都逐渐增加,移植30min后达到高峰,WM-G可明显提高肝移植受体存活率,明显降低AST及ALT,WM-G移植15min后肝细胞中LC3-II的表达低于UW-G;超微结构显示移植15min后WM-G肝细胞中早期自体吞噬性液泡(AVi)、晚期自体吞噬性液泡(AVd)、自噬溶酶体/溶酶体(包括致密体)的比例明显低于UW-G(P<0.01)。结论自噬参与了肝脏冷缺血再灌注损伤过程,抑制自噬可降低肝移植术后冷缺血再灌注损伤。
