大鼠缺血再灌注移植肾组织中Calpain、cPLA2的表达及高压氧对其的影响  

Expression of Calpain and cPLA2 in rat s kidney after ischemia-reperfusion and the effects of hyperbaric oxygen therapy

作　　者：宋先初 谢智慧 梁国标 暴丁溯 庄永祥 王欣 梁天才 杜洋 李浩 杜江 Song Xianchu;Xie Zhihui;Liang Guobiao;Bao Dingshuo;Zhuang Yongxiang;Wang Xin;Liang Tiancai;Du Yang;Li Hao;Du Jiang(Department of Urology,the Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;Department of Hyperbaric Oxygen,the Affiliated Hospital of Zunyi Medical Unirersiry,Zunyi 563000,China;Department of Urology,the Traditional Chinese Medicine Affiliated Hospital of Southwest Medical University,Luzhou 030699,China;Department of Urology,the First People’s Hospital of Putian City,Putian 351199,China)

机构地区：[1]遵义医科大学附属医院泌尿外科,563000 [2]遵义医科大学附属医院高压氧科,563000 [3]西南医科大学附属中医医院泌尿外科,泸州030699 [4]莆田市第一人民医院泌尿外科,351199

出　　处：《国际泌尿系统杂志》2021年第5期871-876,共6页International Journal of Urology and Nephrology

基　　金：国家自然科学基金(81460135);贵州省科技基金(SY3149-2012);贵州省教育厅创新群体重大研究项目(KY045-2017)。

摘　　要：目的通过观察同种大鼠缺血再灌注(IRI)移植肾组织中钙离子依赖性蛋白酶(Calpain)及胞浆型磷脂酶A2(cPLA2)的表达情况,同时观察高压氧(HBO)治疗对Calpain、cPLA2表达的影响,初步探讨细胞凋亡的内质网通路及丝裂原活化蛋白激酶(MAPK)通路在肾HU发生中的作用及HBO治疗对通路的影响。方法同种大鼠移植肾石蜡标本分为假手术组、肾移植组和肾移植+HB0治疗组,分别以移植后1、3和5 h为时间点各分为三个小组。分别应用HE染色法观察病理改变,免疫组织化学(IHC)检测组织中Calpain、cPLA2蛋白的表达情况。结果①肾组织HE染色变化:假手术组肾组织形态基本正常,各时间点组织结构差异不明显;肾移植组各时间点组织损伤较重;HB0治疗组在1 h及3 h时,肾组织病变程度与肾移植组差异较小,在5 h时,组织损伤减轻;②IHC法检测蛋白的表达变化:Calpain和cPLA2均在肾小管上皮细胞胞浆中表达。相对定量分析显示:假手术组中,在各时间点二者的表达水平比较,差异无统计学意义(P>0.05);随再灌注时间延长,二者的表达水平在肾移植组及HB0治疗组中均呈明显上升趋势(P<0.05),且高于假手术组(P<0.05);但在各时间点二者在肾移植组中的表达水平明显高于HB0治疗组(P<0.05)。结论肾移植后早期产生组织损害,再灌注后移植肾组织的Calpain和cPLA2蛋白的表达水平明显增加,随再灌注时间的延长越明显。HB0治疗可明显抑制IRI移植肾组织中Calpain、cPLA2蛋白的表达。Objective To observe the expression of calcium-dependent protease(Calpain)and cytosolic phospholipase A2(cPLA2)in rats kidney after ischemia-reperfusion treatment followed by hyperbaric oxygen(HBO)therapy or not,and to investigate the role of endoplasmic reticulum and mitogen-activated protein kinase(MAFK)pathway of apoptosis in the pathogenesis of renal ischemia-reperfusion injury and the effects of HBO therapy on the pathway.Methods The paraffin-embedded renal tissue were divided into sham group,kidney transplantation group and kidney transplantation+HBO treatment group(HBO treatment group),then they were divided into 3 groups at 1,3 and 5 h after the transplantation,respectively.Hematoxylin and eosin(HE)staining was used to observe the pathological rhanges and the expression of calpain and cPLA2 protein were evaluated by immunohisto-chemistrv(IHC).Results Changes of renal HE staining,sham group,the morphology of renal tissue was basically normal,no significant difference was observed at each time point;kidney transplantation group:the renal damage was more serious at each time point;HBO treatment group:compared with the kidney transplantation group,there was no significant difference at 1 h and 3 h,however the renal damage was alleviated at 5 h.Immunohistochemistry(IHC)was used to evaluated the expression change of proteins:Both Calpain and cPLA2 were expressed in the cytoplasm of renal tubular epithelial cells.Relative quantitative analysis showed that there was no significant difference in expression level of Calpain and cPLA2 at each time point in sham group(P>0.05);with the extension of reperfusion time,the expression of both Calpain and cPLA2 were significantly increased in kidney transplantation group and HBO treatment group(P<0.05);however,their expression levels in kidney transplantation group were significantly hight*r than that in the HBO treatment group(P<0.05).Conclusions Tissue damage occurred in the early stage after renal transplantation,and the expression levels of calpain and cPLA2 proteins in

关 键 词：肾移植 再灌注损伤 大鼠 模型 动物 钙离子依赖性蛋白酶 

分 类 号：R699.2[医药卫生—泌尿科学]