天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (1): 25-29.doi: 10.3969/j.issn.0253-9896.2015.01.007

• 细胞与分子生物学 • 上一篇    下一篇

重组人红细胞生成素对高糖诱导人肾小管上皮细胞增殖及凋亡的影响及其可能机制

陈艳霞， 吴险峰， 房向东△， 秦晓华， 黄翀， 涂卫平#br#   

  1. 江西省南昌大学第二附属医院肾脏内科（邮编 330006）
  • 收稿日期:2014-07-09 修回日期:2014-08-26 出版日期:2015-01-15 发布日期:2015-01-30
  • 通讯作者: 房向东 E-mail:xiangdongfang818@sina.com
  • 基金资助:
    江西省自然科学基金（20122BAB205006）

Effects of erythropoietin in high glucose induced proliferation and apoptosis of human kidney proximal tubular epithelial cells and the possible mechanism

CHEN Yanxia, WU Xianfeng, FANG Xiangdong△, QIN Xiaohua, HUANG Chong, TU Weiping   

  1. Department of Nephrology, the Second Affiliated Hospital of Medical College of Nanchang University, Nanchang 330006, China
  • Received:2014-07-09 Revised:2014-08-26 Published:2015-01-15 Online:2015-01-30
  • Contact: FANG Xiangdong E-mail:xiangdongfang818@sina.com

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摘要： 摘要： 目的 探讨重组人红细胞生成素（rhEPO）对高糖诱导的正常人肾小管上皮（HK-2）细胞增殖及凋亡的影响及其可能机制。 方法 将体外培养的 HK-2 细胞按随机数字表法分为空白对照组、高糖诱导组（高糖终浓度为 30 mmol/L）、甘露醇对照组（甘露醇浓度为 24.5 mmol/L）、rhEPO 对照组（rhEPO 终浓度为 20 U/mL）、不同浓度 rhEPO 干预组（高糖终浓度为 30 mmol/L +rhEPO 终浓度分别为 5、10、20 U/mL）及 Rho 激酶抑制剂（Y27632）组（Y27632 终浓度为 30 μmol/L+高糖终浓度为 30 mmol/L）， 各组均刺激 24 h。 应用 RT-PCR 法检测各组 HK-2 细胞 RhoA、 ROCK1 mRNA 的表达； MTT 法测定细胞增殖， 流式细胞技术分析细胞凋亡。 结果 高糖诱导组 RhoA 及 ROCK1 mRNA 表达较空白对照组显著升高（P < 0.05）， 不同浓度 rhEPO 干预组 RhoA mRNA 及 ROCK1 mRNA 的表达较高糖诱导组显著减少（P < 0.05）， 高糖诱导组及不同浓度 rhEPO 干预组 RhoA mRNA 与 ROCK1 mRNA 表达呈正相关。 rhEPO 可明显促进 HK-2 细胞增殖（P < 0.05）， 而高糖可诱导正常人肾小管上皮细胞凋亡， 加入不同浓度 rhEPO 或 Y27632 干预后， 其凋亡明显受抑制（P < 0.05）， 且在实验 rhEPO 浓度范围内， rhEPO 促进增殖及抑制凋亡的作用呈现浓度依赖性。 结论 rhEPO 可促进高糖诱导的 HK-2 细胞增殖， 抑制高糖诱导的 HK-2 细胞凋亡， 其机制可能与阻断 RhoA/ROCK 信号通路有关。

关键词: 红细胞生成素, 重组, 细胞增殖, 细胞凋亡, rho 相关激酶类, HK-2 细胞, 高糖, RhoA/ROCK 信号通路

Abstract: Abstract： Objective To study the effects of erythropoietin (rhEPO) in high glucose induced proliferation and apopto⁃ sis of human kidney proximal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells cultured in vitro were divided into several groups randomly: blank control group, high glucose group, mannitol group, rhEPO control group, different concentrations of rhEPO treatment groups (5, 10, 20 U/mL) and Rho kinase group. The reverse tran⁃ scription polymerase chain reaction (RT-PCR) was used to evaluate the mRNA levels of RhoA and ROCK after 24 hours. Tetrazolium salt method (MTT) was used to determine the cell proliferation. Cell apoptosis was detected by flow cytometry. Results Compared with blank control group the expression levels of RhoA and ROCK1 mRNA were significantly in⁃ creased in high glucose group (P < 0.05). RhoA, ROCK1 mRNA expressions significantly decreased in rhEPO group than those of high glucose group (P < 0.05). There was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in high glucose group and rhEPO group. MTT method showed that rhEPO significantly promoted the prolifer⁃ ation of HK-2 cells (P < 0.05). Flow cytometry analysis showed that high glucose induced apoptosis in HK-2 cells, which was significantly inhibited in rhEPO group and Rho kinase group as compared to that of high glucose group in a concentra⁃ tion dependent manner (P < 0.05). Conclusion rhEPO can promote HK-2 cell proliferation and inhibit apoptosis, which may be related to RhoA/ROCK signaling pathway.

Key words: erythropoietin, recombinant, cell proliferation, apoptosis, rho-associated kinases, HK- 2 cells, high glu? cose;RhoA/ROCK signaling pathway