Effect of transfection of recombinant human endothelial nitric oxide synthase gene on hypertrophic scar fibroblasts in vitro.
- Author:
Ping YANG
1
;
Ai-li WANG
;
De-wu LIU
;
Shun XU
;
Yao-hui GU
;
Jing HUANG
;
Bo CHEN
;
Qian-cheng LUO
;
Qing JIA
;
Zhi-hong WU
Author Information
- Publication Type:Journal Article
- MeSH: Cicatrix, Hypertrophic; metabolism; Collagen Type I; genetics; metabolism; Collagen Type III; metabolism; Fibroblasts; metabolism; Humans; In Vitro Techniques; Nitric Oxide; metabolism; Nitric Oxide Synthase Type III; genetics; RNA, Messenger; metabolism; Transfection
- From: Chinese Journal of Burns 2008;24(4):275-277
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the feasibility of transfection of recombinant human endothelial nitric oxide synthase (eNOS) into human hypertrophic scar fibroblasts (HSFbs), and to observe NO secretion and the synthesis of collagen I and III.
METHODSRecombinant human eNOS with karyocyte expressive vector was constructed in vitro, then was transfected into HSFbs which was isolated from hypertrophic scar tissues and cultured in vitro (T group). The HSFbs untransfected (normal culture) or transfected with empty-vector was used as control group and empty-vector group respectively. The mRNA expression of eNOS, collagen I and III was determined by Realtime PCR. The content of NO was determined by NO assay kit.
RESULTSThe expression of eNOS mRNA in T group was 5.92 +/- 0.21, which was obviously higher than that in empty-vector group (0.98 +/- 0.13, P < 0.05). The expression of collagen I mRNA (0.76 +/- 0.15), and collagen III (0.79 +/- 0.08) in T group was significantly lower than those in empty-vector group (0.98 +/- 0.15, 1.02 +/- 0.12, P < 0.05, respectively). The content of NO in T group (36.1 +/- 0.8 micromol/L) was obviously higher than that in empty-vector group (28.4 +/- 1.0 micromol/L, P < 0.01) and control group (27.7 +/- 1.3 micromol/L, P < 0.01).
CONCLUSIONHSFbs can be the target cells for eNOS gene transfection. The transfected cells can express eNOS and produce NO, which inhibit the synthesis of collagen.