天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (2): 133-137.doi: 10.11958/59025

• 细胞与分子生物学 • 上一篇    下一篇

Roscovitine在衣霉素诱导足细胞损伤中的保护作用

高翔 1,2， 张悦 3， 张爱金 1， 付芃 1， 李佳琳 1， 吴建 1， 刘巍 1△   

  1. 1河北医科大学病理学教研室 （邮编 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
  • 收稿日期:2015-06-02 修回日期:2015-09-17 出版日期:2016-02-15 发布日期:2016-02-15
  • 通讯作者: △通讯作者 E-mail： lwei929@126.com E-mail:lwei929@126.com
  • 基金资助:
    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）

The role of roscovitine in tunicamycin induced podocyte injury

GAO Xiang1, 2, ZHANG Yue3, ZHANG Ai′jin1, FU Peng1, LI Jialin1, WU Jian1, LIU Wei1△   

  1. 1 Department of Pathology, Hebei Medical University, Shijiazhuang 050017, China; 2 Department of Vascular Surgery, the Second Hospital of Hebei Medical University, 3 Department of Diagnostics, Hebei Medical University
  • Received:2015-06-02 Revised:2015-09-17 Published:2016-02-15 Online:2016-02-15
  • Contact: △Corresponding Author E-mail: lwei929@126.com E-mail:lwei929@126.com

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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（1.0 μmol/L， TM） 组， 各实验组分别刺激 3、 6、 12 h。（2） 正常对照组、 Tunicamycin （1.0 μmol/L， TM） 组及 Tunicamycin+Rosco⁃ vitine（20、 40 μmol/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、 40 μmol/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。

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

Abstract: Objective To observe the protective effects of roscovitine on the podocyte injury induced by endoplasmic reticulum stress (ERS) caused by tunicamycin. Methods The differentiated podocytes cultured at 37 ℃ were randomly divided into: (1) Control group, DMSO group and tunicamycin group (TM, 1.0 μmol/L). The treatment was given for 3, 6 and 12 hours in three groups. (2) For control group, tunicamycin group, tunicamycin+roscovitine group (20, 40 μmol/L, TM+ROS), the treatment was given for 12 hours. The podocyte apoptosis was detected by flow cytometry and TUNEL method. The expressions of Cdk5, 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 Cdk5, GRP78, Caspase-12 and CHOP were also up-regulated significantly in TM group (P < 0.05). (2) Flow cytometry 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 Cdk5, can reduce the podocyte apoptosis induced by tunicamycin. The protective effects of roscovitine on podocytes can be a novel approach of treating diabetic nephropathy.

Key words: podocyte, apoptosis, Tunicamycin, Cdk5, endoplasmic reticulum stress, Roscovitine