肝移植,自体%低氧诱导因子α%c-Jun氨基末端激酶%Caspase-3%缺血再灌注损伤
肝移植,自體%低氧誘導因子α%c-Jun氨基末耑激酶%Caspase-3%缺血再灌註損傷
간이식,자체%저양유도인자α%c-Jun안기말단격매%Caspase-3%결혈재관주손상
Liver transplantation,autologous%Hypoxia inducible factor-1α%c-Jun N-terminal kinase%Caspase-3%Ischemia reperfusion injury
目的 检测低氧诱导因子α(HIF-1α)在自体原位肝移植大鼠肝脏的表达情况,探讨HIF-1α在JNK信号转导通路中的作用机制.方法 采用单纯随机抽样法将96只SD大鼠分为正常对照组、自体肝移植组、低氧预适应+自体肝移植组,每组32只.正常对照组:仅开腹分离血管,不作其他处理;自体肝移植组:采用经门静脉灌注法建立稳定的70%自体原位肝移植模型;低氧预适应+自体肝移植组:术前用8%氮氧混合气体低氧预处理90min,其他处理同自体肝移植组.分别于术后1、2、12、24h检测大鼠血清ALT、AST水平,免疫组织化学染色法测定大鼠肝脏组织HIF-1α的表达,RT-PCR检测c-Jun mRNA表达水平,Western blot检测Cleaved Caspase-3蛋白表达水平.多组比较采用方差分析,两组比较采用t检验.结果 术后1、2、12、24 h,低氧预适应+自体肝移植组大鼠血清ALT和AST水平显著低于自体肝移植组,但高于正常对照组,差异有统计学意义(F=2631.371、1177.642、810.383、682.848,743.618、1095.522、375.995、580.613,P＜0.05).自体肝移植组大鼠术后各时相点HIF-1α蛋白的表达水平明显低于低氧预适应+自体肝移植组,但均高于正常对照组,差异有统计学意义(F=191.737,284.482,459.419,213.782,P＜0.05).低氧预适应+自体肝移植组大鼠术后1、2、12 h c-Jun mRNA表达水平显著低于自体肝移植组,但均高于正常对照组,差异有统计学意义(F=66.211,53.169,9.645,P＜0.05),术后24 h c-Jun mRNA表达水平3组间比较,差异无统计学意义(F=1.100,P＞0.05).低氧预适应+自体肝移植组大鼠术后各时相点Cleaved Caspase-3蛋白表达水平显著低于自体肝移植组,但均高于正常对照组,差异有统计学意义(F=23.133,31.158,14.347,29.043,P＜0.05).结论 肝移植大鼠低氧预适应后HIF-1α的高表达可能通过抑制JNK信号通路的持续激活,下调Cleaved Caspase-3蛋白的表达,从而抑制肝细胞凋亡,减轻肝脏缺血再灌注损伤.
目的 檢測低氧誘導因子α(HIF-1α)在自體原位肝移植大鼠肝髒的錶達情況,探討HIF-1α在JNK信號轉導通路中的作用機製.方法 採用單純隨機抽樣法將96隻SD大鼠分為正常對照組、自體肝移植組、低氧預適應+自體肝移植組,每組32隻.正常對照組:僅開腹分離血管,不作其他處理;自體肝移植組:採用經門靜脈灌註法建立穩定的70%自體原位肝移植模型;低氧預適應+自體肝移植組:術前用8%氮氧混閤氣體低氧預處理90min,其他處理同自體肝移植組.分彆于術後1、2、12、24h檢測大鼠血清ALT、AST水平,免疫組織化學染色法測定大鼠肝髒組織HIF-1α的錶達,RT-PCR檢測c-Jun mRNA錶達水平,Western blot檢測Cleaved Caspase-3蛋白錶達水平.多組比較採用方差分析,兩組比較採用t檢驗.結果 術後1、2、12、24 h,低氧預適應+自體肝移植組大鼠血清ALT和AST水平顯著低于自體肝移植組,但高于正常對照組,差異有統計學意義(F=2631.371、1177.642、810.383、682.848,743.618、1095.522、375.995、580.613,P＜0.05).自體肝移植組大鼠術後各時相點HIF-1α蛋白的錶達水平明顯低于低氧預適應+自體肝移植組,但均高于正常對照組,差異有統計學意義(F=191.737,284.482,459.419,213.782,P＜0.05).低氧預適應+自體肝移植組大鼠術後1、2、12 h c-Jun mRNA錶達水平顯著低于自體肝移植組,但均高于正常對照組,差異有統計學意義(F=66.211,53.169,9.645,P＜0.05),術後24 h c-Jun mRNA錶達水平3組間比較,差異無統計學意義(F=1.100,P＞0.05).低氧預適應+自體肝移植組大鼠術後各時相點Cleaved Caspase-3蛋白錶達水平顯著低于自體肝移植組,但均高于正常對照組,差異有統計學意義(F=23.133,31.158,14.347,29.043,P＜0.05).結論 肝移植大鼠低氧預適應後HIF-1α的高錶達可能通過抑製JNK信號通路的持續激活,下調Cleaved Caspase-3蛋白的錶達,從而抑製肝細胞凋亡,減輕肝髒缺血再灌註損傷.
목적 검측저양유도인자α(HIF-1α)재자체원위간이식대서간장적표체정황,탐토HIF-1α재JNK신호전도통로중적작용궤제.방법 채용단순수궤추양법장96지SD대서분위정상대조조、자체간이식조、저양예괄응+자체간이식조,매조32지.정상대조조:부개복분리혈관,불작기타처리;자체간이식조:채용경문정맥관주법건립은정적70%자체원위간이식모형;저양예괄응+자체간이식조:술전용8%담양혼합기체저양예처리90min,기타처리동자체간이식조.분별우술후1、2、12、24h검측대서혈청ALT、AST수평,면역조직화학염색법측정대서간장조직HIF-1α적표체,RT-PCR검측c-Jun mRNA표체수평,Western blot검측Cleaved Caspase-3단백표체수평.다조비교채용방차분석,량조비교채용t검험.결과 술후1、2、12、24 h,저양예괄응+자체간이식조대서혈청ALT화AST수평현저저우자체간이식조,단고우정상대조조,차이유통계학의의(F=2631.371、1177.642、810.383、682.848,743.618、1095.522、375.995、580.613,P＜0.05).자체간이식조대서술후각시상점HIF-1α단백적표체수평명현저우저양예괄응+자체간이식조,단균고우정상대조조,차이유통계학의의(F=191.737,284.482,459.419,213.782,P＜0.05).저양예괄응+자체간이식조대서술후1、2、12 h c-Jun mRNA표체수평현저저우자체간이식조,단균고우정상대조조,차이유통계학의의(F=66.211,53.169,9.645,P＜0.05),술후24 h c-Jun mRNA표체수평3조간비교,차이무통계학의의(F=1.100,P＞0.05).저양예괄응+자체간이식조대서술후각시상점Cleaved Caspase-3단백표체수평현저저우자체간이식조,단균고우정상대조조,차이유통계학의의(F=23.133,31.158,14.347,29.043,P＜0.05).결론 간이식대서저양예괄응후HIF-1α적고표체가능통과억제JNK신호통로적지속격활,하조Cleaved Caspase-3단백적표체,종이억제간세포조망,감경간장결혈재관주손상.
Objective To investigate the expression of hypoxia inducible factor-1α(HIF-1α)in the transplanted liver in rats and the role of HIF-1α in the JNK signaling pathway.Methods Ninety-six SD rats were randomly divided into normal control(NC)group,autotransplantation(AT)group,hypoxia preconditioning (HP)+ AT group according to simple random sampling method.No treatment was applied to the rats in the NC group except for blood vessel separation.Stable rat models of 70％ AT was established in the AT group.Rats in the HP + AT group were given 8％ oxygen mixed gas for 90 minutes before the operation.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected at 1,2,12,24 hours after operation,and the expression of HIF-1α in the hepatic tissue,mRNA expression of c-Jun,and protein expression of Cleaved Caspase-3 were detected by immunohistochemical staining,reverse transcription-polymerase chain reaction and Western blot,respectively.All data were analyzed using the analysis of variance or t test.Results The levels of ALT and AST in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2,12 and 24 hours after the operation(F=2631.371,1177.642,810.383,682.848;743.618,1095.522,375.995,580.613,P ＜0.05).The protein expression levels of HIF-1α in the AT group were significantly lower than those in the HP + AT group,but higher than those in the NC group at 1,2,12 and 24 hours after operation(F = 191.737,284.482,459.419,213.782,P ＜ 0.05).The mRNA expression levels of c-Jun in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2 and 12 hours after operation(F = 66.211,53.169,9.645,P ＜ 0.05).There was no significant difference in the mRNA expression of c-Jun among the 3 groups at 24 hours after operation(F = 1.100,P ＞ 0.05).The protein expressions of Cleaved Caspase-3 in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2,12 and 24 hours after the operation(F =23.133,31.158,14.347,29.043,P ＜ 0.05).Conclusion High expression of HIF-1α after HP inhibits apoptosis of hepatic cells and alleviates ischemia reperfusion injury of hepatic tissues by suppressing JNK activation,down-regulating protein expression of Cleaved Caspase-3 after orthotopic liver transplantation in rats.
