CD73在小鼠肝缺血再灌注损伤内源性保护机制中的作用及其与TGF-β_(1)/Smad3信号通路的关系  

Role of CD73 in endogenous protective mechanism of hepatic ischemia-reperfusion injury in mice and the relationship with TGF-β_(1)/Smad3 signaling pathway

作　　者：户占飞 章艳君 郭浩 王丽[1] 喻文立[2] 杜洪印[2] Hu Zhanfei;Zhang Yanjun;Guo Hao;Wang Li;Yu Wenli;Du Hongyin(First Central Clinic College,Tianjin Medical University,Tianjin 300192,China;Department of Anesthesiology,Tianjin First Center Hospital,Tianjin 300192,China)

机构地区：[1]天津医科大学一中心临床学院,天津300192 [2]天津市第一中心医院麻醉科,天津300192

出　　处：《中华麻醉学杂志》2022年第5期595-599,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81700659,81700569);天津市自然科学基金(17JCYBJC28000);天津市卫生行业重点攻关项目(16KG101)。

摘　　要：目的评价胞外-5′-核苷酸酶(CD73)在小鼠肝缺血再灌注损伤内源性保护机制中的作用及其与转化生长因子β_(1)(TGF-β_(1))/SMAD家族成员3(Smad3)信号通路的关系。方法SPF级健康雄性C57BL/6小鼠24只,8~10周龄,体重20~23 g,采用随机数字表法分为3组(n=8):假手术组(S组)只游离肝门,不阻断;肝缺血再灌注组(IR组)阻断肝门30 min后恢复灌注制备小鼠肝缺血再灌注损伤模型;肝缺血再灌注+CD73特异性抑制剂组(APCP组)腹腔注射CD73特异性抑制剂APCP 40 mg/kg,10 min后行缺血再灌注。于再灌注6 h时采集眶静脉血样,测定血清ALT和AST浓度,随后处死小鼠取肝组织,采用Western blot法检测CD73、TGF-β_(1)和磷酸化Smad3(p-Smad3)表达,ELISA法检测IL-1β和TNF-α含量,可见分光光度计测定MDA含量和SOD活性,光镜下观察肝组织病理学结果。结果与S组比较,IR组和APCP组血清AST和ALT浓度、肝组织IL-1β、TNF-α和MDA含量升高,SOD活性降低,CD73、TGF-β_(1)和p-Smad3表达上调(P<0.05);与IR组比较,APCP组血清AST和ALT浓度、肝组织IL-1β、TNF-α和MDA含量升高,SOD活性降低,CD73、TGF-β_(1)和p-Smad3表达下调(P<0.05)。APCP组肝组织病理学损伤较IR组加重。结论CD73参与了小鼠肝缺血再灌注损伤内源性保护机制的过程,可能与调控TGF-β_(1)/Smad3信号通路有关。Objective To evaluate the role of ecto-5′-nucleotidase(CD73)in endogenous protective mechanism of hepatic ischemia-reperfusion(I/R)injury in mice and the relationship with transforming growth factor beta1(TGF-β_(1))/Smad3 signaling pathway.Methods Twenty-four SPF healthy male C57BL/6 mice,aged 8-10 weeks,weighing 20-23 g,were divided into 3 groups(n=8 each)by the random number table method:sham operation group(S group),hepatic I/R group(IR group)and hepatic I/R plus CD73 specific inhibitor group(APCP group).The hepatic hilum was only exposed but not occluded in group S.The hepatic portal was occluded for 30 min followed by reperfusion to develop the model of hepatic I/R in anesthetized animals in group IR.CD73-specific inhibitor APCP 40 mg/kg was intraperitoneally injected,and 10 min later hepatic I/R was performed.Orbital venous blood samples were collected at 6 h of reperfusion for determination of serum alanine transaminase(ALT)and aspartate transaminase(AST)concentrations.Then the mice were sacrificed,and liver tissues were obtained for determination of the expression of CD73,TGF-β_(1) and phosphorylated Smad3(p-Smad3)(by Western blot),contents of interleukin-1β(IL-1β)and tumor necrosis factor-alpha(TNF-α)(by enzyme-linked immunosorbent assay),and content of malondialdehyde(MDA)and activity of superoxide dismutase(SOD)(with a visible spectrophotometer)and for microscopic examination of the pathological changes of liver tissues(with a light microscope).Results Compared with group S,the concentrations of AST and ALT in serum and contents of IL-1β,TNF-αand MDA in liver tissues were significantly increased,the activity of SOD was decreased,and the expression of CD73,TGF-β_(1) and p-Smad3 was up-regulated in IR and APCP groups(P<0.05).Compared with group IR,the concentrations of AST and ALT in serum and contents of IL-1β,TNF-αand MDA in liver tissues were significantly increased,the activity of SOD was decreased,and the expression of CD73,TGF-β_(1) and p-Smad3 in liver tissues was down-regulated in gr

关 键 词：5′-核苷酸酶 肝 再灌注损伤 转化生长因子Β Smad3蛋白质 

分 类 号：R575[医药卫生—消化系统]