Roscovitine在衣霉素诱导足细胞损伤中的保护作用  

The role of roscovitine in tunicamycin induced podocyte injury

作　　者：高翔[1,2] 张悦[3] 张爱金[1] 付芃[1] 李佳琳[1] 吴建[1] 刘巍[1] 

机构地区：[1]河北医科大学病理学教研室,050017 [2]河北医科大学第二医院血管外科 [3]河北医科大学诊断学教研室

出　　处：《天津医药》2016年第2期133-137,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81400731);河北省自然科学基金资助项目(H2013206139;H2015206257);河北省卫生和计划生育委员会重点科技研究计划项目(20130140);河北医科大学大学生创新性实验计划资助项目(201410089018)

摘　　要：目的应用衣霉素诱导内质网应激(ERS),观察Roscovtine对ERS导致足细胞损伤的保护作用。方法体外培养永生化小鼠足细胞,取37℃分化成熟细胞随机分组:(1)正常对照组、DMSO组及衣霉素Tunicamycin(1.0μmol/L,TM)组,各实验组分别刺激3、6、12 h。(2)正常对照组、Tunicamycin(1.0μmol/L,TM)组及Tunicamycin+Rosco-vitine(20、40μmol/L,TM+ROS)组,各实验组分别刺激12 h。应用流式细胞术及TUNEL法检测足细胞凋亡情况;应用Western blot检测细胞周期素依赖性蛋白激酶5(Cdk5)及ERS标志蛋白GRP78、Caspase-12、CHOP的表达变化。结果 (1)与正常对照组和DMSO组相比,Tunicamycin刺激3、6及12 h后,TM组足细胞凋亡率及Cdk5、GRP78、Caspase-12和CHOP蛋白的表达水平均呈明显的时间依赖性增高(P<0.05);(2)加入Roscovitine干预后,与TM组相比,TM+ROS(20、40μmol/L)组足细胞凋亡率及GRP78、Caspase-12和CHOP蛋白的表达水平均显著降低(P<0.05),其干预作用呈现明显的剂量依赖性。结论 Cdk5抑制剂Roscovitine能显著抑制衣霉素诱导的足细胞凋亡,从而发挥细胞保护作用。Roscovitine的足细胞保护作用可能有助于糖尿病肾病的治疗。Objective To observe the protective effects of roscovitine on the podocyte injury induced by endoplasmic reticulum stress （ERS） caused by tunicamyein. Methods The differentiated podocytes cultured at 37 ℃ were randomly di- vided into： （1） Control group, DMSO group and tunicamyein group （TM, 1.0 pomol/L）. The treatment was given for 3, 6 and 12 hours in three groups. （2） For control group, tunieamyciu group, tunicamycin＋roscovitine group （20, 40 p, mol/L, TM＋ROS）, the treatment was given for 12 hours. The podocyte apoptosis was detected by flow eytometry and TUNEL method. The expressions of CdkS, GRP78, Caspase-12 and CHOP were detected by Western blot assay. Results （1） Compared with control group and DMSO group, the podocyte apoptosis was increased significantly in a time dependent manner after tunicamycin treatment in TM group; the protein expressions of CdkS, GRP78, Caspase-12 and CHOP were also up-regulated significantly in TM group （P 〈 0.05）. （2） Flow eytometry and TUNEL analysis showed that tunicamycin induced apoptosis in podo- cytes, which was significantly inhibited by roscovitine in a concentration dependent manner in TM＋ROS group as compared to that of TM group （P 〈 0.05）. The protein expressions of GRP78, Caspase-12 and CHOP were also significantly decreased in a concentration dependent manner in TM＋ROS group compared to those of TM group （P 〈 0.05）. Conclusion Roscovi- tine, the inhibitor of CdkS, can reduce the podocyte apoptosis induced by tunicamyein. The protective effects of roscovitine on podoeytes can be a novel approach of treating diabetic nephropathy.

关 键 词：足细胞 凋亡 衣霉素 细胞周期素依赖性蛋白激酶5 内质网应激 ROSCOVITINE 

分 类 号：R692.6[医药卫生—泌尿科学]