蛋白激酶Cβ抑制剂LY333531对糖尿病大鼠造影剂肾病的保护作用  

Protective effect of protein kinase Cβ inhibitor LY333531 in diabetic rats with contrast-induced nephropathy

作　　者：蔡学英 殷婷 朱英 刘炳炜 胡炜 Cai Xueying;Yin Ting;Zhu Ying;Liu Bingwei;Hu Wei(Department of Intensive Care Unit,Affiliated Hangzhou First People's Hospital,Zhejiang University School of Medicine,Hangzhou 310006,China;Department of Cardiology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院重症医学科,杭州310006 [2]南京医科大学第一附属医院(江苏省人民医院)心内科,南京210029

出　　处：《中华危重症医学杂志（电子版）》2020年第6期412-418,共7页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：浙江省医药卫生一般研究项目(2015KYA178)。

摘　　要：目的探讨蛋白激酶Cβ(PKCβ)抑制剂LY333531对造影剂诱导的糖尿病大鼠急性肾损伤的保护作用及其机制。方法将40只大鼠分为健康对照组(C组)、糖尿病组(D组)、糖尿病造影剂组(DC组)以及糖尿病+Y333531+造影剂组(DCL组),每组各10只。对D组、DC组及DCL组大鼠腹腔注射2%链脲佐菌素(60 mg/kg)建立糖尿病模型;对DCL组大鼠给予LY333531灌胃预处理(10 mg·kg-1·d-1),持续14 d;对DC组与DCL组大鼠进行尾静脉注射76%复方泛影葡胺(10 ml/kg),建立造影剂肾病模型。苏木素-伊红(HE)染色观察各组大鼠肾组织病理变化;检测各组大鼠血糖、血肌酐、尿微量白蛋白(m Alb)和N-乙酰-β-D-葡萄糖苷酶(NAG)水平;采用实时荧光定量PCR法检测转化生长因子β1(TGFβ1)、Smad3、Smad7、Bax、Caspase3和Bcl2的信使RNA(mRNA)表达水平;采用Western-blotting检测TGFβ1、Bax、Caspase3、Bcl2蛋白及磷酸化PKCβ(pPKCβ)/PKCβ、磷酸化p38(p-p38)/p38的比值。结果HE结果显示,C组大鼠肾小球、肾小管结构正常;D组大鼠肾组织内肾小球及肾小管结构形态较为完整,间质周围极少量炎症细胞;DC组肾小管上皮细胞脱落、周围炎症细胞浸润明显;DCL组大鼠肾小管上皮细胞脱落程度明显降低,肾小管及肾小球周围间质少量炎症细胞浸润。各组大鼠间血糖、血肌酐、尿m Alb、尿NAG水平、TGFβ1蛋白及其m RNA、Smad3 m RNA、Smad7 m RNA、Bax蛋白及其m RNA、Caspase3蛋白及其m RNA、Bcl2蛋白及其m RNA表达水平、p PKCβ/PKCβ和p-p38/p38比值间比较,差异均有统计学意义(F=67.976,P<0.001;F=27.155,P<0.001;F=41.201,P<0.001;F=59.635,P<0.001;F=21.073,P<0.001;F=28.365,P<0.001;F=15.215,P<0.001;F=36.273,P<0.001;F=14.489,P<0.001;F=23.172,P<0.001;F=17.103,P<0.001;F=29.916,P<0.001;F=12.026,P<0.001;F=13.368,P<0.001;F=6.126,P=0.002;F=6.434,P=0.002)。进一步两两比较发现,D组、DC组以及DCL组大鼠的血糖水平较C组大鼠均明显升高(P均<0.05);与DC组大鼠比较,DCL组大�Objective To explore the protective effect of protein kinase Cβ(PKCβinhibitor LY333531 in diabetic rats with contrast-induced acute kidney injury and its mechanism.Methods Totally 40 rats were randomly divided into a control group(C group),a diabetic group(D group),a diabetes with contrast-induced nephropathy group(DC group)and a diabetes with Y333531 and contrast-induced nephropathy group(DCL group),10 rats in each group.The rats in the D,DC and DCL groups were injected intraperitoneally with 2%streptozotocin(60 mg/kg)to induce a diabetes model.Then the rats in the DCL group were pretreated with LY333531(10 mg·kg-1·d-1)by intragastric administration for 14 d;the rats in the DC and DCL groups were injected with 76%diatrizoate(10 mL/kg)by tail vein to establish a model of contrast-induced nephropathy.The pathological changes in kidney were observed by hematoxylin-eosin(HE)staining.The levels of blood glucose,serum creatinine,urine microalbumin(mAlb)and urine Nacetyl-β-D-glucosidase(NAG)were detected.The messenger RNA(mRNA)levels of transforming growth factor beta1(TGFβ1),Smad3,Smad7,Bax,Caspase3 and Bcl2 were examined by realtime fluorescence quantitative PCR.The protein levels of TGFβ1,Bax,Caspase3 and Bcl2 were detected by Western-blotting,as well as the phospho-PKCβ(pPKCβ)/PKCβand phospho-p38(pp38)/p38.Results According to HE,the glomeruli and renal tubules in the group C were structurally normal;the glomeruli and renal tubules in the group D were relatively structurally complete with few inflammatory cells around the interstitium;the epithelial cells of renal tubules in the DC group were exfoliated and the inflammatory cells obviously infiltrated around renal tubules;in the DCL group,the degree of epithelial cell shedding in renal tubules was significantly reduced,and a small amount of inflammatory cells infiltrated in the interstitium around renal tubules and glomeruli.The blood glucose,serum creatinine,urine mAlb,urine NAG,TGFβ1 protein and its mRNA,Smad3 mRNA,Smad7 mRNA,Bax protein and its mRNA,C

关 键 词：糖尿病 造影剂肾病 蛋白激酶C 大鼠 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学] R-332[医药卫生—临床医学]