Up-regulation by human recombinant transforming growth factor β-1 of collagen production in cultured dermal fibroblasts is mediated by the inhibition of nitric oxide signaling
Up-regulation by human recombinant transforming growth factor β-1 of collagen production in cultured dermal fibroblasts is mediated by the inhibition of nitric oxide signaling
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DOI：
10.1016/s1072-7515(98)00303-2
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发表时间：
1999-03-01
影响因子：
5.2
通讯作者：
Prasad, JK
中科院分区：
文献类型：
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作者：
Chu, AJ;Prasad, JK
背景：增生性瘢痕仍然是烧伤幸存者最致残的后遗症。其发病机制尚不清楚。然而，在烧伤增生性瘢痕（HS）中一直观察到胶原积累。研究设计：我们研究了源自HS的真皮成纤维细胞中胶原蛋白的产生，该细胞已经发展了9个月至2年。进行重建手术以去除培养成纤维细胞的HS。同样，正常细胞从患者的供体部位（DS）生长，提供自体移植到HS部位。在含有5%胎牛血清(含l -抗坏血酸（100 μ g/mL）和β -氨基腈（100 μ g/mL）的微量必需氨基酸培养基中，通过监测条件培养基中20 h [H-3]脯氨酸掺入细菌胶原酶iii -可消化蛋白中，比较和分析HS和DS成纤维细胞胶原蛋白的生成。结果：我们没有发现HS和DS在体外产生胶原蛋白方面有明显差异。无论来自HS还是DS的成纤维细胞，经过3天的预处理，人重组转化生长因子β -1 (tgf - β 1) （20 ng/mL）显著刺激了胶原蛋白的产生，大约刺激了250%。相比之下，硝普钠（SNP）在100 μ M处表现出显著的抑制作用（68%），被血红蛋白（10 μ M）拯救。tgf - β 1显著降低一氧化氮（NO）生成55%。相比之下，SNP治疗后NO水平急剧上升350%。表皮生长因子对胶原蛋白生成和一氧化氮水平均无影响。线性回归分析显示NO水平与胶原生成呈显著负相关（r = 0.72; p < 0.05），提示NO信号参与了胶原生成的调节。与这一观点一致，我们进一步表明，n -硝基- l -精氨酸甲酯（100 μ M）在TGF -1和SNP存在下分别对胶原生成产生协同刺激和抑制作用。8-BrcGMP （300 μ M）模拟NO抑制作用，亚甲基蓝（50 μ M）恢复被SNP抑制的胶原生成。此外，8-BrcGMP抵消了胶原生成的刺激。结论：HS培养的真皮成纤维细胞在胶原生成及对调控的反应方面与正常真皮成纤维细胞无明显差异。胶原蛋白产生的抑制是通过cgmp依赖性NO作用实现的。TGF β -1抑制NO/cGMP信号，确保其对真皮成纤维细胞胶原生成的刺激作用。[J]中华口腔外科杂志1999；18(1):71- 80。(C) 1999年由美国外科医师学会出版)。
Background: Hypertrophic scarring remains the most disabling sequela for burn survivors. Little is known about its pathogenesis. Collagen accumulation, however, has been consistently observed in burn hypertrophic scars (HS).Study Design: We have studied collagen production in the dermal fibroblasts derived from HS, which has developed for 9 months to 2 years. Reconstructive surgery was performed to remove HS from which the fibroblasts were cultured. Similarly, the normal cells were grown from the patient's donor site (DS), which provided autografting to the HS site. Collagen production in HS and DS fibroblasts was compared and analyzed in minimal essential amino acid medium containing 5% fetal bovine serum with inclusion of L-ascorbic acid (100 mu g/mL) and beta-aminopropoinitrile (100 mu g/mL) by monitoring a 20-h [H-3]proline incorporation into bacterial collagenase III-digestible protein in the conditioned media.Results: We failed to detect any significant difference in collagen production in vitro between HS and DS. Irrespective of the fibroblasts from HS or DS, collagen production was substantially stimulated by human recombinant transforming growth factor beta-1 (TGF-beta 1) (20 ng/mL) by approximately 250% after a 3-day pretreatment. In contrast, sodium nitroprusside (SNP) at 100 mu M exhibited significant suppression (68%), which was rescued by hemoglobin (10 mu M). TGF-beta 1 significantly decreased nitric oxide (NO) production by 55%. In contrast, NO level drastically increased by 350% following SNP treatment. Epidermal growth factor showed no effect on either collagen production or NO level. The linear regression analysis shows a significant inverse correlation (r = 0.72; p < 0.05) of NO level with collagen production, suggesting the involvement of NO signaling in the modulation of collagen production. Consistent with the notion, we further showed that N-nitro-L-arginine methyl ester (100 mu M) caused a synergistic stimulation and an arrested inhibition of collagen production in the presence of TGF beta-1 and SNP, respectively. 8-BrcGMP (300 mu M) mimicked the NO inhibitory action, while methylene blue (50 mu M) restored the collagen production which was inhibited by SNP. Moreover, 8-BrcGMP offset the stimulation of collagen production.Conclusions: The dermal fibroblasts derived from HS were not different from normals with respect to collagen production and their responses to regulations. The inhibition of collagen production was achieved by a cGMP-dependent NO action. TGF beta-1 inhibited NO/cGMP signaling to ensure its stimulatory effect on collagen production in the dermal fibroblasts. (J Am Coil Surg 1999;188:271-280. (C) 1999 by the American College of Surgeons).