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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) preconditioning on hepatic ischemia-reperfusion (IR) injury in mice and its possible mechanism. Methods: One hundred male ICR mice were randomly divided into 4 groups: Sham group, 70% hepatic ischemia-reperfusion group, Saline group and Tβ4 group. The serum of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected at 1, 3, 6, 12 and 24 h after ischemia / reperfusion. The liver tissues collected at the above time points were divided into two parts: one part was fixed with formaldehyde for HE staining to observe the pathological changes of liver tissues; the other part was preserved in liquid nitrogen for Western blot and PCR experiments. The levels of Tβ4, AKT, p-AKT, Bad and p-Bad were detected by Western blot. The mRNA levels of TNF-α and IL-6 were detected by PCR. Results: Compared with normal liver tissue, the expression level of Tβ4 decreased at 1 h and reached the normal level at 6 h after ischemia / reperfusion. The levels of ALT and AST in Tβ4 group were significantly lower than those at 70% after reperfusion The expression of p-AKT and p-Bad in Tβ4 group was significantly lower than that in Tβ4 group (P <0.05), and the pathological changes in liver tissue of Tβ4 group were significantly improved. The expression of TNF-α and IL- There was no difference between the groups at 12 h after ischemia-reperfusion. Conclusion: Thymosin β4 can reduce the hepatic ischemia-reperfusion injury in mice by directly activating the AKT-Bad signaling pathway and inhibiting the expression of inflammatory cytokines.