他克莫司预处理对大鼠肝脏缺血再灌注损伤的影响

Tacrolimus pretreatment on liver ischemia reperfusion injury in rats

目的 研究他克莫司（FK506）预处理对大鼠肝脏缺血再灌注（IR）损伤的影响.方法 将32只SD大鼠随机分为4组,分别为假手术组（S组）、缺血再灌注组（IR组）、FK506低剂量组（L组）、FK506高剂量组（H组）.缺血60 min,再灌注6h取材,比较各组大鼠血清ALT、AST水平;利用ELISA法检测血清炎症因子TNF-α、IL-1β水平;采用HE染色观察肝组织病理改变;通过实时荧光定量PCR （RT-qPCR）、免疫组织化学法及免疫印迹法观察FK506预处理对高迁移率族蛋白B1（HMGB1）表达的影响.结果 与IR组相比,L组和H组大鼠血清ALT[（424.0±137.4） U/L、（291.0 ±42.0） U/L]、AST[（554.2±127.7） U/L、（410.2±7.0） U/L]及炎症因子TNF-α[（115.1±49.0） ng/L、120.4±28.5）ng/L]、IL-1 β[（424.5±105.2） ng/L、（612.1 ±49.6） ng/L]水平均显著降低（P＜0.05）.L组和H组较IR组肝血窦淤血、肝细胞坏死及炎症细胞浸润均明显减轻.L组和H组中HMGB1的mRNA及蛋白表达显著低于IR组,但高于S组（P＜0.01）.上述观察指标在L组和H组之间比较差异无统计学意义（P＞0.05）.结论 FK506预处理可以显著降低大鼠肝脏IR损伤后HMGB1的表达,抑制炎症因子释放,减少细胞坏死,从而减轻肝脏IR损伤.

Objective To determine the effects of tacrolimus （FK506） pretreatment on the liver ischemia reperfusion （IR） injury.Methods 32 mature SD rats were randomly assigned into four groups,which were sham-operated group （S）,ischemia reperfusion group （IR）,low-dose FK506-treated group （L） and high-dose FK506-treated group （H）.After the treatment of liver ischemia for 60 minutes and reperfusion for 6 hours,the levels of serum ALT and AST in rats were tested.The TNF-α and IL-1β levels were evaluated by enzyme linked immunosorbent assay.Liver damage was assessed by paraffin sections stained with H＆E.The quantitative real-time PCR,the immunohistochemistry and Western blot were used to detect the expression of HMGB1 mRNA and protein with or without FK506 pretreatment.Results The levels of serum ALT [（424.0 ± 137.4）U/L,（291.0 ±42.0）U/L],AST [（554.2 ± 127.7）U/L,（410.2 ±7.0）U/L],TNF-α [（115.1±49.0）ng/L,120.4±28.5） ng/L] and IL-1β [（424.5 ±105.2） ng/L,（612.1 ± 49.6） rig/L] decreased markedly in the group L and group H compared with the group IR （P 〈 0.05）.The liver in the IR group showed hepatic sinusoids congestion,neutrophil infiltration and necrosis.In contrast,tissue damage of the L group and the H group was significantly decreased.The expressions of HMGB1 mRNA and protein reduced significantly when pretreatment with FK506 after reperfusion （P 〈 0.01）.However,there was no significant difference between the group L and group H （P 〉 0.05）.Conclusion FK506 pretreatment can protect the liver by reducing the expression of HMGB1,inhibiting the release of inflammatory cytokines and alleviating cell necrosis after the liver ischemia reperfusion injury in rats.