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[重组人核心蛋白聚糖对转化生长因子-β1刺激胶原晶格中纤维母细胞的拮抗作用]

[The antagonistic effect of recombinant human decorin on TGF-beta1 stimulation of fibroblasts in collagen lattices].

作者信息

Zhang Zhi, Li Xiao-jian, Liang Da-rong, Li Ye-yang, Xu Wei-shi

机构信息

Department of Burns and Plastic Surgery, Guangzhou Red Cross Hospital, Guangzhou 510220, PR China.

出版信息

Zhonghua Shao Shang Za Zhi. 2006 Jun;22(3):207-10.

Abstract

OBJECTIVE

To mimic contact pattern between decorin and TGF-beta1, in vivo, and investigate the antagonistic effect of recombinant human decorin on TGF-beta1 stimulation of hypertrophic scar fibroblasts in collagen lattices.

METHODS

Fibroblasts populated collagen lattices (FPCL) model was adopted in the study, and they were divided into control group, decorin group [2mg/L recombinant human decorin (rh-decorin) was administered to FPCL], TGF-beta1 group (5 microg/LTGF-beta1 was administered to the culture medium), and TGF-beta1 + decorin group (2mg/L rh-decorin was administered to FPCL, then culture medium containing 5 microg/L TGF-beta1 was added into FPCL). Changes in PAI-1 and alpha-SMA protein expression in scar fibroblasts in collagen lattices were detected with Western blotting at 12 post-administration hour (PAH), 24 PAH, 48 PAH, and 72 PAH, and expressions of PAI-1 and alpha-SMA mRNA were concomitantly examined by RT-PCR.

RESULTS

The contraction of FPCL at each time-point in control group was obviously attenuated compared with that in decorin group, but it was significantly intensified compared with that in TGF-beta1 group. The expression of PAI-1 and alpha-SMA mRNA and protein in TGF-beta1 group (3482 +/- 211, 4320 +/- 272, 0.89 +/- 0.15, 0.56 +/- 0. 11) were markedly increased than those in control group (1764 +/- 147, 1699 +/- 146, 0.29 +/- 0.06, 0.21 +/- 0.06, P < 0.01), while no obvious difference of them was found between control and other two groups.

CONCLUSION

Stimulation of scar fibroblasts by TGF-beta1, can be suppressed when rh-decorin is blended into collagen lattices, indicating that decorin is effective in neutralizing TGF-beta1 in vitro. The pathogenesis of hypertrophic scar might be related to up-regulation of TGF-beta1 with the lack of decorin after cutaneous injury.

摘要

目的

在体内模拟核心蛋白聚糖与转化生长因子-β1(TGF-β1)之间的接触模式,研究重组人核心蛋白聚糖对TGF-β1刺激胶原凝胶中增生性瘢痕成纤维细胞的拮抗作用。

方法

采用成纤维细胞填充胶原凝胶(FPCL)模型,将其分为对照组、核心蛋白聚糖组[向FPCL中加入2mg/L重组人核心蛋白聚糖(rh-核心蛋白聚糖)]、TGF-β1组(向培养基中加入5μg/L TGF-β1)和TGF-β1+核心蛋白聚糖组(向FPCL中加入2mg/L rh-核心蛋白聚糖,然后向FPCL中加入含5μg/L TGF-β1的培养基)。在给药后12小时(PAH)、24 PAH、48 PAH和72 PAH,用蛋白质印迹法检测胶原凝胶中瘢痕成纤维细胞中纤溶酶原激活物抑制剂-1(PAI-1)和α-平滑肌肌动蛋白(α-SMA)蛋白表达的变化,并用逆转录聚合酶链反应(RT-PCR)同时检测PAI-1和α-SMA mRNA的表达。

结果

对照组FPCL在各时间点的收缩与核心蛋白聚糖组相比明显减弱,但与TGF-β1组相比明显增强。TGF-β1组中PAI-1和α-SMA mRNA及蛋白的表达(3482±211、4320±272、0.89±0.15、0.56±0.11)明显高于对照组(1764±147、1699±146、0.29±0.06、0.21±0.06,P<0.01),而对照组与其他两组之间未发现明显差异。

结论

当rh-核心蛋白聚糖混入胶原凝胶中时,可抑制TGF-β1对瘢痕成纤维细胞的刺激,表明核心蛋白聚糖在体外能有效中和TGF-β1。增生性瘢痕的发病机制可能与皮肤损伤后TGF-β1上调而核心蛋白聚糖缺乏有关。

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