PKC抑制剂对力竭运动大鼠肾脏裂孔膜蛋白表达的影响  

Effect of PKC inhibitor on renal podocyte slit diaphragm protein expression in exhausted rats

作　　者：安静芳 李航 杨帆 谢文杰 李鹏飞 刘姣 崔迪 周刚[1] AN Jing-fang;LI Hang;YANG Fan;XIE Wen-jie;LI Peng-fei;LIU Jiao;CUI Di;ZHOU Gang(College of Physical Education,Hunan University,Changsha 410000,China)

机构地区：[1]湖南大学体育学院,长沙410082

出　　处：《中国应用生理学杂志》2022年第2期126-131,共6页Chinese Journal of Applied Physiology

基　　金：湖南省体育局自然科学基金资助(2020XH020)。

摘　　要：目的:观察大鼠在一次性力竭运动后肾脏裂孔膜蛋白的表达水平,探究PKC抑制剂对其蛋白表达水平的影响,揭示PKC在运动性蛋白尿形成中的作用机制。方法:SD雄性大鼠30只随机分为对照组(C)、运动组(E)、运动联合PKC抑制剂组(EPI),每组10只。E组和EPI组大鼠分别进行一次性跑台力竭运动(25 m/min),EPI组大鼠运动前1 d及1 h腹腔注射PKC抑制剂白屈菜红碱(chelerythrine, 5 mg/kg),C组和E组注射相应体积的生理盐水。运动后即刻麻醉后,取血液、尿液及肾脏组织,使用化学比色法检测尿蛋白、尿酸、尿糖、血尿素、血尿酸、血糖水平,使用荧光探针法检测肾脏ROS水平,使用Western blot法检测肾脏PKC、Nox2、Nox4、nephrin、podocin蛋白表达。结果:(1)与C组相比,E组尿蛋白、尿酸、尿糖、血尿素、血尿酸显著增多(P<0.05),血糖显著减少(P<0.01),肾脏ROS生成显著增多(P<0.01),肾脏nephrin、podocin蛋白表达明显降低(P<0.05),PKC、Nox2、Nox4蛋白表达明显增多(P<0.05);(2)与E组比,EPI组尿蛋白、尿糖、血尿素显著减少(P<0.05),血糖显著增加(P<0.01),肾脏ROS生成显著降低(P<0.01),EPI组肾组织中nephrin、podocin蛋白表达明显增加(P<0.05),PKC、Nox2蛋白表达明显降低(P<0.05)。结论:一次性力竭运动通过PKC/NOX/ROS途径使大鼠肾脏裂孔膜蛋白nephrin、podocin表达下调;PKC抑制剂缓解力竭运动导致的肾脏裂孔膜蛋白表达下降,预防运动性蛋白尿的发生。Objective:To investigate the expression level of podocyte slit diaphragm protein in rats after one-time exhaustive exercise,to explore the effect of PKC inhibitor on the protein expression level,and to reveal the mechanism of PKC in the formation of exercise-induced proteinuria.Methods:Thirty male SD rats were randomly divided into control group(C),exercise group(E)and exercise combining with PKC inhibitor group(EPI),with 10 rats in each group.Rats in group E and EPI performed one single bout of exhaustive exercise(25 m/min),rats in group EPI were intraperitoneally injected with a PKC inhibitor(chelerythrine,5 mg/kg)1 day and 1 hour before exercise respectively,while rats in group C and E were injected with the same volume of saline.Results:①Compared with group C,the levels of urine protein,uric acid,urine sugar,blood urea,and blood uric acid of rats in group E were increased significantly(P<0.05),the level of blood glucose was reduced significantly(P<0.01),and renal ROS production was increased significantly(P<0.01).The expressions of nephrin and podocin protein in renal tissue were decreased significantly(P<0.05),while the expressions of PKC,Nox2,and Nox4 protein were increased significantly(P<0.05).②Compared with group E,the levels of urinary protein,urine glucose and blood urea in EPI group were decreased significantly(P<0.05),the level of blood glucose was increased significantly(P<0.01),renal ROS production was reduced significantly(P<0.01).the expressions of nephrin and podocin protein in renal tissues of the EPI group were increased significantly(P<0.05),while the expressions of PKC and Nox2 protein was reduced significantly(P<0.05,P<0.01).Conclusion:One-time exhaustive exercise can down-regulate the expressions of nephrin and podocin through PKC/Nox/ROS pathways in the kidney of rats;PKC inhibitor alleviates the decrease in the expression of podocyte slit diaphragm protein caused by exhaustive exercise,and prevents the occurrence of exercise-induced proteinuria.

关 键 词：运动性蛋白尿 蛋白激酶C(PKC) NADPH氧化酶(Nox) 活性氧(ROS) NEPHRIN PODOCIN 

分 类 号：G804.5[文化科学—运动人体科学]