中国血液净化

中国血液净化 ›› 2020, Vol. 19 ›› Issue (10): 683-687.doi: 10.3969/j.issn.1671-4091.2020.10.009

• 基础研究 • 上一篇    下一篇

转化生长因子-RhoGTP 酶／Rho 激酶系统信号通路在低氧诱导大鼠肾小球内皮细胞-间充质细胞转分化中的作用及机制研究

马金兰1,孙雪1,张志强1,王燕1,梅峰1   

  1. 1青海大学附属医院肾病内科
  • 收稿日期:2020-01-15 修回日期:2020-07-20 出版日期:2020-10-12 发布日期:2020-10-12
  • 通讯作者: 梅峰 meifengqh@163.com E-mail:meifengqh@163.com
  • 基金资助:
    青海科技厅基础研究项目(2016-ZJ-709)

The role and mechanism of transforming growth factor-RhoGTPase/Rho Kinase signaling pathway in hypoxia- induced trans- differentiation of rat glomerular endothelial cells

  1. 1Department of Nephrology, Qinghai University Affiliated Hospital, Xining 810001, China
  • Received:2020-01-15 Revised:2020-07-20 Online:2020-10-12 Published:2020-10-12

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摘要： 【摘要】目的  低氧对大鼠肾小球内皮细胞(rat glomerular endothelial cells，rGECs) 转化生长因子(transforming growth factor，TGF）-β1的表达和转分化相关蛋白表达的影响及机制研究。方法  低氧培养箱培养rGECs，根据低氧培养时间将细胞随机分为5 组，检测细胞增殖活性、TGF-β1及转分化相关蛋白表达情况；在低氧培养细胞中在合适的时间分别加入RhoA/ROCK 阻滞剂和TGF-β1受体阻滞剂，分别再次测定：①细胞培养液中TGF-β1表达；②转分化相关蛋白表达；③RhoA/ROCK 活性变化。结果①随着低氧培养时间的延长，rGECs 增殖活性增加，72h 活性最大(F=546.63，P＜0.001)；②与常氧组比较，低氧组的TGF- β1(F=16.320,P＜0.001) 和α- SMA(F =5.032,P=0.009) 表达升高，而CD- 31(F=9.882,P＜0.001)表达降低；③经RhoA/ROCK 阻滞剂和TGF-β1受体阻滞剂阻滞后α-SMA 蛋白表达下降(相对表达量由1.423 分别下降至0.750、0.434), CD-31 蛋白表达升高 ( 相对表达量由0.741 分别上升至
0.779、0.934)。结论研究发现低氧可致rGECs 增殖活性增加，促进rGECs 向间质细胞转分化。并通过细胞信号阻滞剂研究发现低氧可能通过TGF-β1-RhoA/ROCK信号通路促进rGECs 向间质细胞转分化。

关键词: 低氧, 大鼠肾小球内皮细胞, 内皮细胞-间充质细胞转分化, 转化生长因子-RhoA/ROCK 信号通路

Abstract: 【Abstract】Purpose To investigate the effect and mechanism of hypoxia on the expression of TGF-β1 and the proteins relating to endothelial-to-mesenchymal transition (EndMT) in rat glomerular endothelial cells (rGECs). Methods rGECs were cultured in a hypoxia incubator, and the cells were randomly divided into 5 groups according to hypoxia treatment time. Cell proliferation activity, the expression of TGF-β1 and EndMT related proteins were then measured in the 5 cell groups. RhoA/ROCK blocker and TGF-β1 receptor blocker were added to the hypoxia treated cells at appropriate time, and then the following indicators were assayed: ①TGF-β1 in cell culture medium; ②TGF-β1 and EndMT related proteins; ③change of RhoA/ROCK activity. Results ①The proliferative activity of rGECs increased with the prolongation of hypoxia treatment time with the biggest activity after hypoxia treatment for 72 hours (2.040±0.110, F=546.63, P＜0.001)；②Compared with the normoxic group, the expressions of TGF-β1 (F=16.320, P＜0.001) and α-SMA (F=5.032, P=0.009) in the hypoxic group increased significantly, while CD-31 protein (F=9.882, P＜0.001) decreased significantly. ③Under the hypoxic circumstances and treated with RhoA/ROCK blocker and TGF-β1 blocker, α-SMA protein decreased (relative amount from 1.423 to 0.750 and 0.434 respectively) and CD-31 protein increased (relative
amount from 0.741 to 0.779 and 0.934 respectively). Conclusion Hypoxia can increase cell proliferation and promote EndMT of rGECs. The use of TGF-β1 and RhoA/ROCK blockers under hypoxic conditions suggest that hypoxia environment promotes EndMT of rGECs via the TGF-β1-RhoA/ROCK signaling pathway.

Key words: Hypoxia, Rat glomerular endothelial cells, Endothelial-to-mesenchymal transition, TGF-β1-RhoA/ROCK signaling pathway

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