Inhibition effect of small interfering RNA of connective tissue growth factor on the expression of vascular endothelial growth factor and connective tissue growth factor in cultured human peritoneal mesothelial cells  

作　　者：LIU Fu-you XIAO Li PENG You-ming DUAN Shao-bin LIU Hong LIU Ying-hong LING Gui-hui YUAN Fang CHEN Jun-xiang FU Xiao ZHU Jian-lian 

机构地区：[1]Division of Nephrology, Second Affiliated Xiangya Hospital, Central South University, Changsha 410002, China

出　　处：《Chinese Medical Journal》2007年第3期231-236,共6页中华医学杂志（英文版）

基　　金：the National Natural Science Foundation of China(No. 30370811)

摘　　要：Background The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor β1 （TGF-β1 ） plays a key role in regulating tissue repair and remodelling after injury. Connective tissue growth factor （CTGF）, a downstream mediator of TGF-β1 inducing fibrosis, has been implicated in peritoneal fibrosis. Vascular endothelial growth factor （VEGF） plays a key role in angiogenesis that can hasten peritoneal fibrosis. In this study, we investigated the effect of small interfering RNA （siRNA） of CTGF by pRETRO-SUPER （PRS） retrovirus vector on the expression of CTGF and VEGF in human peritoneal mesothelial cells. Methods Retrovirus producing CTGF siRNA were constructed from the inverted oligonucleotides and transferred into packaging cell line PT67 with lipofectamine, and the virus supernatant was used to infect human peritoneal mesothelial cell （HPMC）. The cells were divided into seven groups： low glucose DMEM, low glucose DMEM ＋ TGF-β1 5 ng/ml, low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS-CTGF-siRNA1-4 and low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS. The expression of CTGF and VEGF were measured by semiquantitative RT-PCR and Western blot. Results Low levels of CTGF and VEGF were detected in confluent HPMCs. Following stimulation with TGF-β1 , the levels of CTGF and VEGF were significantly upregulated （P〈0.01）. Introduction of PRS-CTGF-siRNA1-4 resulted in the significant reduction of CTGF mRNA and protein, and VEGF mRNA （P〈0.01）, especially in groups PRS-CTGF-siRNA, and PRS-CTGF-siRNA4. The introduction of PRS void vector did not have these effects （P〉0.05）. Conclusions The expression of CTGF siRNA mediated by PRS retrovirus vector can effectively reduce the level of CTGF and VEGF induced by TGF-β1 in cultured HPMCs. This study may provide potential therapeutic strategies to prevent the peritoneal fibrosis.对腹分离的腹膜反应能导致的背景纤维变性。(TGF-1 ) 转变生长因素 1 在调整织物修理并且在损害以后改变起一个关键作用。结缔组织生长因素(CTGF ) ，导致纤维变性的 TGF-1 的一个下游的调停人，在腹纤维变性被含有。脉管的 endothelial 生长因素(VEGF ) 在能赶快的 angiogenesis 起一个关键作用腹纤维变性。在这研究，我们调查了 CTGF 的小介入 RNA ( siRNA )的效果由pRETRO超级( PRS )在人的腹的 CTGF 和 VEGF 的表示上的 retrovirus 向量生产 CTGF siRNA 的间皮的 cells.Methods Retrovirus 从转换 oligonucleotides 被构造并且转了进与 lipofectamine 包装房间线 PT67 ，并且病毒上层清液被用来感染人的腹间皮的房间( HPMC )。房间被划分成七个组：低葡萄糖 DMEM，低葡萄糖 DMEM + TGF-1 5 ng/ml，低葡萄糖 DMEM + TGF-1 5 ng/ml + PRS-CTGF-siRNA1-4 和低葡萄糖 DMEM + TGF-1 5 ng/ml + PRS。CTGF 和 VEGF 的表示被 semiquantitative RT-PCR 和 CTGF 的西方的 blot.Results 底层测量， VEGF 在汇合的 HPMC 被检测。有 TGF-1 的后面的刺激， CTGF 和 VEGF 的层次是显著地 upregulated (P0.01 ) 。PRS-CTGF-siRNA1-4 的介绍导致了 CTGF 的重要减小 mRNA 和蛋白质，和 VEGF mRNA (P0.01 ) ，特别在组 PRS-CTGF-siRNA1 和 PRS-CTGF-siRNA4。PRS 空向量的介绍没有这些效果(P0.05 ).Conclusions CTGF siRNA 的表示由 retrovirus 向量能有效地减少的 PRS 调停了 CTGF 和 VEGF 的水平在有教养的 HPMC 由 TGF-1 导致了。这研究可以提供潜在的治疗学的策略阻止腹纤维变性。

关 键 词：peritonealfibrosis connective tissue growth factor vascular endothelial growth factor RNA  small interfering 

分 类 号：R459.5[医药卫生—治疗学] R692.5[医药卫生—临床医学]