他克莫司预处理对大鼠自体原位肝移植冷缺血再灌注损伤的影响和机制

The effect and mechanism of tacrolimus pretreatment on cold ischemia reperfusion injury of transplanted liver in rats

目的分析他克莫司预处理对自体原位肝移植大鼠肝脏冷缺血再灌注损伤(IRI)的影响及机制。方法无特殊病原体雄性Sprague Dawley大鼠24只,8~10周龄,220~250 g,采用随机区组法将大鼠分3组,每组8只。假手术组仅开腹关腹,他克莫司组术前经静脉注射他克莫司,模型组注射等量生理盐水。他克莫司组和模型组制备自体原位肝移植冷IRI模型。检测再灌注后大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β),HE染色评估肝组织损伤。实时荧光定量PCR和蛋白质电泳检测低氧诱导因子-1α(HIF-lα)、血红素加氧酶-1(HO-1)相对表达。结果模型组大鼠血清ALT为(1332.0±52.8)U/L、AST为(2472.0±257.8)U/L,高于假手术组(65.0±17.4)U/L、(222.3±45.2)U/L和他克莫司组(789.9±54.0)U/L、(533.4±31.6)U/L,差异均有统计学意义(均P<0.05)。他克莫司组肝组织损伤程度轻于模型组。模型组大鼠血清TNF-α、IL-1β高于假手术组和他克莫司组,差异均有统计学意义(均P<0.05)。他克莫司组HIF-lα、HO-1相对表达均高于模型组,差异均有统计学意义(均P<0.05)。结论他克莫司预处理通过增加HIF-1α和HO-1表达,抑制炎症因子生成,减轻大鼠自体原位肝移植冷IRI。

Objective To investigate the roles of tacrolimus pretreatment on hepatic ischemia reperfusion injury(IRI)and its possible mechanism in a rat autologous orthotopic liver transplantation(AOLT)model.Methods For 24 specific pathogen free 8-10 week male Sprague Dawley rats(220-250g)were randomly and equally divided into three groups.The abdomen of sham-operated group was only opened and closed;the treatment with tacrolimus was administered via dorsal penile vein before the experiment in tacrolimus-pretreated group;the AOLT group and tacrolimus-pretreated group were set to construct the AOLT IRI rat models.The levels of ALT,AST,tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in serum were tested after the reperfusion.The change of liver structure was evaluated by H&E staining.The quantitative real-time PCR(RT-qPCR)and Western blot assay were used to test the mRNA and protein level of hypoxia-inducible factor-1α(HIF-1α)and heme oxygenase-1(HO-1).Results The levels of serum ALT(1332.0±52.8)U/L and AST(2472.0±257.8)U/L in the AOLT group were higher than the levels in the sham-operated group(65.0±17.4)U/L,(222.3±45.2)U/L and tacrolimus-pretreated group(789.9±54.0)U/L,(533.4±31.6)U/L.The differences were significant(P<0.05).And in the tacrolimus-pretreated group there were less lesions in the liver than in the AOLT group.The serum level of TNF-αand IL-1βof the AOLT group were increased than the sham-operated group and tacrolimus-pretreated group,and the differences were significant(P<0.05).Compared with the AOLT group,the expressions of HIF-1αand HO-1 were increased significantly after the tacrolimus pretreatment(P<0.05).Conclusion Tacrolimus pretreatment could reduce rats hepatic cold IRI by inducing the expressions of HIF-1αand HO-1,and inhibiting the production of inflammatory cytokines.