沉默BCAT1对大鼠腹腔感染性脓毒症急性肾损伤的干预作用  

Interventional effect of silencing BCAT1 on acute kidney injury in rats with peritoneal sepsis

作　　者：石新芳 王育民[1] 张凯秀[1] 崔国峰[2] 何小磊 SHI Xin-fang;WANG Yu-min;ZHANG Kai-xiu;CUI Guo-feng;HE Xiao-lei(Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia O10050,China;不详)

机构地区：[1]内蒙古医科大学附属医院疾病预防控制部,内蒙古呼和浩特010050 [2]内蒙古医科大学附属医院心内科,内蒙古呼和浩特010050 [3]呼和浩特迪安医学检验实验室,内蒙古呼和浩特010010

出　　处：《中华医院感染学杂志》2023年第23期3527-3531,共5页Chinese Journal of Nosocomiology

基　　金：内蒙古自治区科技计划基金资助项目(2020YFDS0043)。

摘　　要：目的探究支链氨基酸转氨酶1(BCAT1)基因沉默在大鼠腹腔感染性脓毒症急性肾损伤中的作用。方法采用随机数字法将80只大鼠随机分成四组:对照组、感染组、感染组+空白质粒组(NC-shRNA)、感染组+BCAT1干扰质粒组(BCAT1-shRNA),采用盲肠结扎穿孔的方法制备大鼠腹腔感染性脓毒症模型,用全自动生化分析仪检测血清肌酐(SCr)、尿素氮(BUN)水平。造模成功后采用NC-shRNA转染NC-shRNA组大鼠,BCAT1-shRNA转染BCAT1-shRNA组大鼠;采用酶联免疫吸附测定法(ELISA)检测大鼠血清中的白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)表达水平;实时荧光定量聚合酶链式反应法(qRT-PCR)和蛋白印迹法检测大鼠血清及肾脏组织中BCAT1和C-反应蛋白(CRP)mRNA表达水平。结果与对照组相比感染组各组大鼠SCr、BUN水平升高(P<0.05),炎症因子TNF-α、IL-1β及IL-6水平升高以及BCAT1和CRP基因和蛋白表达水平升高(P<0.05);与感染组和NC-shRNA组相比,BCAT1-shRNA组大鼠体内TNF-α、IL-1β、IL-6以及BCAT1和CRP基因、蛋白表达水平降低。结论沉默BCAT1基因沉默可以抑制CRP表达,抑制炎症反应,对腹腔感染脓毒症急性肾损伤具有一定治疗作用。OBJECTIVE To explore the role of branched-chain amino acid transaminase 1(BCAT1)gene silencing in acute kidney injury in rats with abdominal infection sepsis.METHODS Eighty rats were randomly divided into four groups by using random number method,including the control group,model group,model group+blank plasmid group(NC-shRNA)and model group+BCAT1 interference plasmid group(BCAT1-shRNA).A rat model of intra-abdominal infectious sepsis was established by cecal ligation and perforation,and serum creatinine(SCr)and blood urea nitrogen(BUN)levels were detected by automatic biochemical analyzer.After successful modeling,the rats in the NC-shRNA group were transfected with NC-shRNA,and the rats in the BCAT1-shRNA group were transfected with BCAT1-shRNA;the expression levels of serum IL-1β,TNF-α and IL-6 in each group were detected by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction and western blot methods were used to detect the expression levels of BCAT1 and CRP mRNA in serum and kidney tissue of rat.RESULTS Compared with the control group,the blood SCr and BUN levels of the rats in the model group were increased(P<0.05).The levels of inflammatory factors TNF-α,IL-1β and IL-6 were increased,and the mRNA expression levels of BCAT1 and CRP were increased(P<0.05).Compared with the model group and NC-shRNA group,the protein levels of TNF-α,IL-1β,IL-6 and mRNA levels of BCAT1 and CRP in the BCAT1-shRNA group were decreased.CONCLUSION BCAT1 gene silencing can inhibit CRP mRNA expression and inflammatory response,which can have a certain therapeutic effect on acute kidney injury in abdominal infection sepsis.

关 键 词：支链氨基酸转氨酶1 基因沉默 急性肾损伤 Nod样受体蛋白3 炎症反应 

分 类 号：R459.7[医药卫生—急诊医学]