TRPM2-CnA-Drp1通路在丙泊酚减轻小鼠肝缺血再灌注致肾损伤中的作用  

Role of TRPM2-CnA-Drp1 pathway in propofol-induced reduction of renal injury induced by hepatic ischemia-reperfusion in mice

作　　者：刘云霞[1] 于洪丽[1] 朱敏[1] 喻文立[1] Liu Yunxia;Yu Hongli;Zhu Min;Yu Wenli(Department of Anesthesiology,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津市第一中心医院麻醉科,300192

出　　处：《中华麻醉学杂志》2021年第6期680-684,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(82072219);2018年天津医学会麻醉学分会中青年科研培育基金项目(TJMZJJ-2018-04);天津市卫生健康委员会科技项目(ZC20052);2018天津市自然科学基金面上项目(18JCYBJC27500)。

摘　　要：目的:评价瞬时受体电位M2(TRPM2)-钙调磷酸酶A(CnA)-线粒体动力相关蛋白1(Drp1)通路在丙泊酚减轻小鼠肝缺血再灌注致肾损伤中的作用。方法:SPF级雄性C57BL6小鼠24只,8周龄,体重20~23 g,采用随机数字表法分为4组(n=6):假手术组(S组)、肝缺血再灌注组(IR组)、丙泊酚组(P组)和TRPM2激动剂腺苷二磷酸核糖(ADPR)+丙泊酚组(AP组)。采用夹闭肝左、中叶门静脉和肝动脉60 min恢复灌注的方法制备肝缺血再灌注损伤模型。P组于造模前1 h时腹腔注射生理盐水0.2 ml,造模前30 min时腹腔注射1%异丙酚30 mg/kg;AP组于造模前1 h时腹腔注射ADPR 10 mg/kg(溶于0.2 ml生理盐水中),造模前30 min腹腔注射1%异丙酚30 mg/kg;S组和IR组分别于造模前1 h和30 min时腹腔注射等体积生理盐水。于再灌注6 h时摘眼球采血检测血清BUN、Cr、ALT和AST水平,然后处死小鼠取肾组织,透射电镜下观察线粒体超微结构,并计算线粒体长径,采用Western blot法检测TRPM2、CnA、磷酸化Drp1(p-Drp1)Ser637和cleaved caspase-3的表达。结果:与S组比较,IR组、P组和AP组血清BUN和Cr浓度升高,肾组织TRPM2、CnA和cleaved caspase-3表达上调,p-Drp1 Ser637表达下调,线粒体长径缩短(P<0.05);与IR组比较,P组血清BUN和Cr浓度降低,肾组织TRPM2、CnA和cleaved caspase-3表达下调,p-Drp1 Ser637表达上调,线粒体长径延长(P<0.05),线粒体损伤减轻,血清ALT和AST浓度差异无统计学意义,AP组血清BUN和Cr浓度差异无统计学意义(P>0.05);与P组比较,AP组血清BUN和Cr浓度升高,肾组织TRPM2、CnA和cleaved caspase-3表达上调,p-Drp1 Ser637表达下调,线粒体长径缩短(P<0.05),线粒体损伤加重。结论:丙泊酚减轻小鼠肝缺血再灌注致肾损伤的机制与其抑制肾组织TRPM2的表达,降低胞内CnA的水平,抑制Drp1 Ser637去磷酸化有关。Objective To evaluate the role of transient receptor potential melastatin 2(TRPM2)-calcineurin A(CnA)-dynamin-related protein 1(Drp1)pathway in propofol-induced reduction of renal injury induced by hepatic ischemia-reperfusion(I/R)in mice.Methods Twenty-four SPF male C57BL6 mice,aged 8 weeks,weighing 20-23 g,were divided into 4 groups(n=6 each)using a random number table method:sham operation group(group S),hepatic I/R group(group IR),propofol group(group P)and TRPM2 agonist(ADPR)combined with propofol group(AP group).Hepatic I/R injury was induced by occluding the portal vein and hepatic artery supplying the left and middle lobes of the liver for 60 min followed by reperfusion in anesthetized rats.In group P,0.2 ml normal saline was injected intraperitoneally at 1 h before establishing the model and 1%propofol 30 mg/kg was injected intraperitoneally at 30 min before establishing the model.In group AP,ADPR 10 mg/kg(in 0.2 ml of normal saline)was injected intraperitoneally at 1 h before establishing the model,and 1%propofol 30 mg/kg was injected intraperitoneally at 30 min before establishing the model.The equal volume of normal saline was given intraperitoneally at 1 h and at 30 min before establishing the model in group S and group IR.Blood samples were taken from the eyeballs for determination of the levels of serum urea nitrogen(BUN),creatinine(Cr),aminotransferase(ALT)and aspartate aminotransferase(AST)at 6 h of reperfusion.The animals were then sacrificed and the kidney tissues were taken,the ultrastructure of myocardial mitochondria was observed using transmission electron microscopy,the average diameter of mitochondria was calculated,and the expression of TRPM2,CnA,phospho-Drp1 Ser637(p-Drp1 Ser637)and cleaved caspase-3 was detected(by Western blot).Results Compared with group S,the concentrations of serum BUN and Cr were significantly increased,the expression of TRPM2,CnA and cleaved caspase-3 in kidney tissues was up-regulated,the expression of p-Drp1 ser637 was down-regulated,and the average diameter of

关 键 词：二异丙酚 再灌注损伤 肝 急性肾损伤 TRPM阳离子通道 钙神经素 线粒体蛋白质类 

分 类 号：R614[医药卫生—麻醉学]