中国血液净化

›› 2008, Vol. 7 ›› Issue (4): 203-206.

• 基础研究 • 上一篇    下一篇

PAMAM纳米载体介导pCTGF-shRNA抑制小鼠腹膜间皮细胞CTGF的表达

易斌 刘伏友 刘虹   

  1. 中南大学湘雅二医院肾内科
  • 收稿日期:2008-01-17 修回日期:1900-01-01 出版日期:2008-04-12 发布日期:2008-04-12
  • 通讯作者: 易斌

Inhibition of CTGF expression in mouse peritoneal mesothelial cells by nano-carrier PAMAM mediated pCTGF-shRNA

YI Bin, LIU Fu-you, LIU Hong, PENG You-ming, LIU Ying-hong.   

  1. 1Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha 410013, China
  • Received:2008-01-17 Revised:1900-01-01 Online:2008-04-12 Published:2008-04-12

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摘要： 目的 研究G9 PAMAM纳米载体介导的结缔组织生长因子（CTGF）短发夹RNA（shRNA）质粒（pCTGF-shRNA）对小鼠腹膜间皮细胞CTGF表达的影响。方法 设计并构建质粒pCTGF1-shRNA、pCTGF2-shRNA和阴性对照pHK-shRNA；应用G9 PAMAM将质粒分别转染4.25%葡萄糖刺激下的第3代小鼠腹膜间皮细胞；采用逆转录多聚酶链式反应（RT-PCR）检测细胞CTGF mRNA水平，采用Western Blot检测CTGF蛋白含量。结果 4.25%葡萄糖可刺激小鼠腹膜间皮细胞CTGF的mRNA和蛋白表达明显上调。pCTGF1-shRNA转染组和pCTGF2-shRNA转染组的CTGF表达较高糖组明显下调（P<0.05），尤其以pCTGF2-shRNA组显著。pHK-shRNA转染组与高糖组比较，差异无显著性（P>0.05）。结论 G9 PAMAM纳米载体可介导pCTGF-shRNA转染原代小鼠腹膜间皮细胞，并能抑制高糖诱导的CTGF表达上调，提示PAMAM介导的pCTGF-shRNA可有效地用于腹膜纤维化的基因治疗

关键词: 结缔组织生长因子, 腹膜间皮细胞, PAMAM, shRNA

Abstract: Objectives To investigate the effects of nano-carrier G9 PAMAM mediated plasmid containing the CTGF-shRNA (pCTGF-shRNA) on the expression of CTGF in mouse peritoneal mesothelial cells. Methods The plasmids named pCTGF1-shRNA、pCTGF2-shRNA and negative control pHK-shRNA were designed and formed. Transfection of plasmids to MPMCs (3rd generation, induced by 4.25% glucose) was performed using G9 PAMAM. The expression of CTGF mRNA was detected by RT-PCR. The protein level of CTGF was determined by Western Blot. Results With the treatment of 4.25% glucose, the expression of CTGF in MPMCs were significantly up-regulated. Introduction of pCTGF1-shRNA and pCTGF2- shRNA resulted in significant reduction of CTGF expression (P<0.05) with pCTGF2-shRNA more effective. But the introduction of pHK-shRNA did not lead to significant reduction in CTGF expression（P>0.05）. Conclusions Nano-carrier G9 PAMAM has the ability to efficiently transfect pCTGF-shRNA into primary MPMCs and has prominent inhibition effects on increased expression of CTGF induced by high glucose. PAMAM vector mediated pCTGF-shRNA can be used as a very efficient strategy in gene therapy of peritoneal fibrosis

Key words: Peritoneal mesothelial cells, PAMAM, shRNA

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