[Effect of transfection of recombinant human endothelial nitric oxide synthase gene on hypertrophic scar fibroblasts in vitro].
OBJECTIVE
To 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.


METHODS
Recombinant 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.


RESULTS
The 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).


CONCLUSIONS
HSFbs can be the target cells for eNOS gene transfection. The transfected cells can express eNOS and produce NO, which inhibit the synthesis of collagen.
Yang
Ping
P
Department of Burns and Plastic Surgery, The 7th People Hospital of Shanghai, Shanghai 200137, PR China.
Wang
Ai-li
AL
Liu
De-wu
DW
Xu
Shun
S
Gu
Yao-hui
YH
Huang
Jing
J
Chen
Bo
B
Luo
Qian-cheng
QC
Jia
Qing
Q
Wu
Zhi-hong
ZH
D017439:Cicatrix, Hypertrophic / Q000378:metabolism*; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection
D024042:Collagen Type I; D024061:Collagen Type III; D012333:RNA, Messenger; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III