胸腺素β4通过激活AKT-Bad信号通路减轻小鼠肝脏缺血再灌注损伤  

Thymosin β4 alleviates mouse liver ischemia-reperfusion injury by activating AKT-Bad pathway

作　　者：封冰[1] 沈健[1] 王猛[1] 唐劲草[1] 李相成[1] 

机构地区：[1]南京医科大学第一附属医院肝脏移植中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2012年第4期449-453,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省"兴卫工程重点人才"基金(RC2007056)

摘　　要：目的 :探讨胸腺素β4(thymosinβ4,Tβ4)预处理对小鼠肝脏缺血再灌注(ischemia-reperfusion,IR)损伤的作用及其可能的机制。方法:100只雄性ICR小鼠随机分为4组:空白对照(Sham)组;70%肝脏缺血再灌注组;生理盐水(Saline)组;Tβ4组。肝脏缺血再灌注后1、3、6、12、24 h收集血清检测丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)水平。上述时间点收集的肝脏组织分为两部分:一部分甲醛固定后进行HE染色观察肝脏组织病理学变化;另一部分保存于液氮中进行Westernblot与PCR实验。Tβ4、AKT、p-AKT、Bad、p-Bad的表达水平用Western blot进行检测;肿瘤坏死因子-α(TNF-α)与白介素-6(IL-6)的mRNA水平用PCR进行检测。结果:与正常肝组织相比,Tβ4表达水平在缺血再灌注后1、3 h降低,从6 h开始恢复至正常水平;再灌注后Tβ4组ALT、AST水平明显低于70%缺血再灌注组与生理盐水组(P<0.05),同时Tβ4组的肝脏组织病理学变化明显改善,TNF-α与IL-6的表达水平也受到抑制;Tβ4组p-AKT与p-Bad的表达水平在缺血再灌注后1、3、6 h升高,从12 h开始各组间无差异。结论:胸腺素β4可以通过直接激活AKT-Bad信号通路和抑制炎症因子的表达减轻小鼠肝脏缺血再灌注损伤。Objective： To investigate the effect of thymosin β4（Tβ4） on mouse hepatic ischemia-reperfusion（IR） injury and its possible mechanism.Methods： One hundred male ICR mice were randomly divided into four groups：sham group,70% IR group,saline group and Tβ4 group respectively.Serum was collected for alanine aminotrasferase（ALT） and aspartate aminotransferase（AST） measurement at 1,3,6,12 and 24 h after reperfusion.Liver tissues were divided into two parts：one part of liver was fixed for HE staining,the other part was maintained in liquid nitrogen for Western blotting and PCR.The expressions of Tβ4,AKT,p-AKT,Bad,p-Bad were performed by Western blotting;the mRNA expressions of TNF-α and IL-6 were detected by PCR.Results： During the process of liver I/R injury,the expression of Tβ4 was decreased at 1,3 h and recovered at 6 h after reperfusion.The levels of ALT and AST in Tβ4 group were lower compared with the 70% IR group and the saline group（P 0.05）,and Tβ4 group also showed alleviated I/R injury as revealed by HE staining.Meanwhile,the expressions of TNF-α and IL-6 were significantly inhibited in Tβ4 group.Furthermore,the Tβ4 group showed elevated phosphorylation levels of AKT（p-AKT） and Bad（p-Bad） at 1,3,6 h after reperfusion.Conclusion： Tβ4 can alleviate hepatic I/R injury through activating AKT-Bad pathway directly and inhibiting the expressions of inflammatory cytokines in mice.

关 键 词：胸腺素Β4 AKT BAD 缺血再灌注损伤 肝脏 

分 类 号：Q26[生物学—细胞生物学]