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目的观察牛磺酸预处理后SD大鼠移植肝脏的白细胞介素-1受体相关激酶-4(interleukin-1 receptor associated kinase-4,IRAK-4)表达水平的变化,探讨牛磺酸预处理抑制肝脏缺血再灌注损害的相关机制。方法雄性SD大鼠128只,完全随机化分为生理盐水对照组(NS组)和牛磺酸预处理组(TA组),每组64只,用Kamada’s袖套法经行肝移植。NS组切取供肝前10 min经腰静脉注射生理盐水1.5 ml,TA组切取供肝前10 min经腰静脉注射牛磺酸(300 mmol/L)1.5 ml。于再灌注后0、60 min及180 min,蛋白免疫印记法及实时荧光定量PCR测定肝组织中IRAK-4 mRNA和蛋白表达水平;凝胶电泳迁移率分析法(EMSA)检测肝组织NF-κB活性及血清TNF-α含量;全自动血清自动生物化学仪检测血清转氨酶;光镜切片检测肝脏组织学。结果牛磺酸预处理能明显提高大鼠1周生存率(P<0.01),改善肝功能,减轻肝组织病理学改变。再灌注后0 min,血清转氨酶、IRAK-4 mRNA与蛋白表达水平、NF-κB活性以及TNF-α含量2组间比较差异无统计学意义(P>0.05);NS组各项指标水平于再灌注60 min出现峰值,但TA组各项指标水平明显低于NS组(P<0.01);再灌注后180 min,2组IRAK-4 mRNA与蛋白表达水平、NF-κB活性以及TNF-α含量均较60 min时有所下降,且TA组下降水平较NS组更为明显(P<0.01)。结论牛磺酸对大鼠移植肝脏的缺血再灌注损伤有明显抑制作用,抑制IRAK-4及其下游的NF-κB、TNF-α表达是牛磺酸预处理减轻肝脏缺血再灌注损害的重要机制之一。
Objective To observe the changes of interleukin-1 receptor associated kinase-4 (IRAK-4) expression in the liver of SD rats after taurine pretreatment, and to explore the pretreatment of taurine. Inhibition of hepatic ischemia-reperfusion injury related mechanisms. Methods 128 male SD rats were completely randomized into normal saline control group (NS group) and taurine pretreatment group (TA group). Each group contained 64 rabbits and were subjected to liver transplantation using Kamada’s cuff method. In the NS group, normal saline (1.5 ml) was injected into the lumbar vein 10 min before the donor’s liver, and in the TA group 10 mL of taurine (300 mmol/L) was injected into the lumbar vein 10 min before the donor liver. Immunohistochemistry and real-time fluorescent quantitative PCR were used to detect the expression of IRAK-4 mRNA and protein in liver tissue at 0, 60 and 180 min after reperfusion. The electrophoretic mobility shift assay (EMSA) was used to detect NF-κB in liver tissue. Activity and serum TNF-α content; automatic serum biochemical analyzer for serum transaminases; light microscopy for liver histology. Results Taurine pretreatment significantly improved the 1-week survival rate (P<0.01), improved liver function, and reduced liver pathological changes. At 0 min after reperfusion, serum aminotransferase, IRAK-4 mRNA and protein expression levels, NF-κB activity, and TNF-α content were not significantly different between the two groups (P>0.05); The peak value appeared at 60 min after infusion, but the levels of each index in TA group were significantly lower than that of NS group (P<0.01). The expression of IRAK-4 mRNA and protein, NF-κB activity and TNF-α content in both groups were 180 min after reperfusion. All decreased compared with 60 minutes, and the level of TA in the TA group was more obvious than that in the NS group (P<0.01). Conclusion Taurine has significant inhibitory effect on ischemia-reperfusion injury of rat liver grafts. The inhibition of IRAK-4 and its downstream expression of NF-κB and TNF-α is taurine pretreatment to reduce hepatic ischemia-reperfusion injury. One of the important mechanisms.