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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

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 i... Full description

Journal Title: Chinese medical journal 2007, Vol.120 (3), p.231-236
Main Author: Liu, Fu-you
Other Authors: 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
Format: Electronic Article Electronic Article
Language: English
Subjects:
Publisher: China: Division of Nephrology, Second Affiliated Xiangya Hospital,Central South University, Changsha 410002, China
ID: ISSN: 0366-6999
Link: https://www.ncbi.nlm.nih.gov/pubmed/17355828
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recordid: cdi_wanfang_journals_zhcmj200703012
title: 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
format: Article
creator:
  • 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
subjects:
  • Animals
  • Base Sequence
  • Cells, Cultured
  • Connective Tissue Growth Factor
  • Epithelial Cells - metabolism
  • Humans
  • Immediate-Early Proteins - analysis
  • Immediate-Early Proteins - antagonists & inhibitors
  • Immediate-Early Proteins - genetics
  • Intercellular Signaling Peptides and Proteins - analysis
  • Intercellular Signaling Peptides and Proteins - genetics
  • Mice
  • Molecular Sequence Data
  • NIH 3T3 Cells
  • Peritoneum - cytology
  • Peritoneum - metabolism
  • Retroviridae - genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • RNA, Messenger - analysis
  • RNA, Small Interfering - pharmacology
  • siRNA
  • Transforming Growth Factor beta1 - pharmacology
  • Vascular Endothelial Growth Factor A - analysis
  • Vascular Endothelial Growth Factor A - genetics
  • 基因表达
  • 结缔组织生长因子
  • 腹膜纤维化
  • 腹膜透析
  • 腹膜间皮细胞
  • 血管内皮生长因子
ispartof: Chinese medical journal, 2007, Vol.120 (3), p.231-236
description: 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.
language: eng
source:
identifier: ISSN: 0366-6999
fulltext: no_fulltext
issn:
  • 0366-6999
  • 2542-5641
url: Link


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titleInhibition 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
creatorLiu, 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
creatorcontribLiu, 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
descriptionBackground 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.
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languageeng
publisherChina: Division of Nephrology, Second Affiliated Xiangya Hospital,Central South University, Changsha 410002, China
subjectAnimals ; Base Sequence ; Cells, Cultured ; Connective Tissue Growth Factor ; Epithelial Cells - metabolism ; Humans ; Immediate-Early Proteins - analysis ; Immediate-Early Proteins - antagonists & inhibitors ; Immediate-Early Proteins - genetics ; Intercellular Signaling Peptides and Proteins - analysis ; Intercellular Signaling Peptides and Proteins - genetics ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Peritoneum - cytology ; Peritoneum - metabolism ; Retroviridae - genetics ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - analysis ; RNA, Small Interfering - pharmacology ; siRNA ; Transforming Growth Factor beta1 - pharmacology ; Vascular Endothelial Growth Factor A - analysis ; Vascular Endothelial Growth Factor A - genetics ; 基因表达 ; 结缔组织生长因子 ; 腹膜纤维化 ; 腹膜透析 ; 腹膜间皮细胞 ; 血管内皮生长因子
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1Xiao, Li
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6Ling, Gui-hui
7Yuan, Fang
8Chen, Jun-xiang
9Fu, Xiao
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0Inhibition 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
1Chinese medical journal
addtitleChinese Medical Journal
descriptionBackground 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.
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8Immediate-Early Proteins - genetics
9Intercellular Signaling Peptides and Proteins - analysis
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15Peritoneum - metabolism
16Retroviridae - genetics
17Reverse Transcriptase Polymerase Chain Reaction
18RNA, Messenger - analysis
19RNA, Small Interfering - pharmacology
20siRNA
21Transforming Growth Factor beta1 - pharmacology
22Vascular Endothelial Growth Factor A - analysis
23Vascular Endothelial Growth Factor A - genetics
24基因表达
25结缔组织生长因子
26腹膜纤维化
27腹膜透析
28腹膜间皮细胞
29血管内皮生长因子
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titleInhibition 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
authorLiu, 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
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7Immediate-Early Proteins - antagonists & inhibitors
8Immediate-Early Proteins - genetics
9Intercellular Signaling Peptides and Proteins - analysis
10Intercellular Signaling Peptides and Proteins - genetics
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14Peritoneum - cytology
15Peritoneum - metabolism
16Retroviridae - genetics
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18RNA, Messenger - analysis
19RNA, Small Interfering - pharmacology
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21Transforming Growth Factor beta1 - pharmacology
22Vascular Endothelial Growth Factor A - analysis
23Vascular Endothelial Growth Factor A - genetics
24基因表达
25结缔组织生长因子
26腹膜纤维化
27腹膜透析
28腹膜间皮细胞
29血管内皮生长因子
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atitleInhibition 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
jtitleChinese medical journal
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abstractBackground 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.
copChina
pubDivision of Nephrology, Second Affiliated Xiangya Hospital,Central South University, Changsha 410002, China
pmid17355828
doi10.1097/00029330-200702010-00012
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