The mechanism of oxythiamine-induced collagen biosynthesis in cultured fibroblasts.
The mechanism of oxythiamine-induced collagen biosynthesis in cultured fibroblasts.
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DOI：
10.1007/s11010-015-2336-z
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发表时间：
2015-05
影响因子：
4.3
通讯作者：
Palka, Jerzy
中科院分区：
文献类型：
--
作者：
Szoka, Lukasz;Karna, Ewa;Palka, Jerzy
氧硫胺(Oxy)是硫胺素的抗维生素。Oxy增加丙酮酸的细胞质浓度，从而促进胶原的生物合成，这一发现使我们研究了这种抗维生素对培养的人皮肤成纤维细胞胶原生物合成的作用机制。用微摩尔浓度(30-1,000µM)的Oxy处理融合成纤维细胞24和48小时。结果发现，与未处理的细胞相比，Oxy依赖的胶原生物合成增加伴随着Pro酶活性和水平的增加。由于磷酸烯醇式丙酮酸(PEP)是胶原生物合成中起重要作用的酶--脯氨酸酶的抑制剂，丙酮酸的相互转化机制被认为是胶原生物合成中的一种调节开关。事实上，磷酸烯醇式丙酮酸羧激酶(PEPCK)的特异性抑制剂3-MPA有助于上调Prolidase的活性，这表明下调PEP的形成是一个潜在的机制。由于α-2-β-1整合素受体和胰岛素样生长因子-I受体激活所诱导的信号调节胶原生物合成和脯氨酸酶活性，因此用免疫印迹法检测这些受体的表达。氧暴露导致胰岛素样生长因子-IR、α-2β-1整合素受体、pERK1/2和核因子-κB p65表达下降。伴随着总ERK1/2表达的增加和Akt蛋白的磷酸化。这些结果提示，氧依赖的人皮肤成纤维细胞胶原生物合成的增加是由于激活了脯氨酸酶的活性和水平，诱导了PAKT的表达，并下调了pERK1/2和已知的胶原基因表达抑制因子-κB p65的表达。
The oxythiamine (OXY) is antivitamin of thiamine. The finding that OXY increases the cytoplasmic concentration of pyruvate, known to enhance collagen biosynthesis, led us to investigate the mechanism of this antivitamin action on collagen biosynthesis in cultured human skin fibroblasts. Confluent fibroblasts were treated with micromolar concentrations (30–1,000 µM) of OXY for 24 and 48 h. It was found that OXY-dependent increase in collagen biosynthesis was accompanied by parallel increase in prolidase activity and level, compared to untreated cells. Since phosphoenolpyruvate (PEP) is known as an inhibitor of prolidase—the enzyme that plays important role in collagen biosynthesis, the mechanism of pyruvate interconversion was considered as a regulatory switch in collagen biosynthesis. In fact, 3-MPA, specific inhibitor of phosphoenolpyruvate carboxykinase (PEPCK), contributed to up-regulation of prolidase activity, suggesting that down-regulation of PEP formation is an underlying mechanism. Since collagen biosynthesis and prolidase activity are regulated by signal induced by activated α2β1 integrin receptor as well as insulin-like growth factor-I receptor (IGF-IR), the expression of these receptors was measured by Western immunoblot analysis. The exposure of the cells to OXY contributed to decrease in IGF-IR, α2β1 integrin receptor, pERK1/2, and NF-κB p65 expressions. It was accompanied by increase in total ERK1/2 expression and induction of phosphorylation of Akt protein. The data suggest that OXY-dependent increase of collagen biosynthesis in cultured human skin fibroblasts results from activation of prolidase activity and level, induction in pAkt expression and down-regulation of pERK1/2 and NF-κB p65, the known inhibitor of collagen gene expression.
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DOI：
10.1083/jcb.147.2.401
发表时间：
1999-10-18
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ivaska J;Reunanen H;Westermarck J;Koivisto L;Kähäri VM;Heino J
通讯作者：
Heino J
影响因子：
4.2
作者：
EMMERSON, KS;PHANG, JM
通讯作者：
PHANG, JM
DOI：
10.1016/0167-4781(90)90037-3
发表时间：
1990-06-21
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
MAKELA, JK;VUORIO, T;VUORIO, E
通讯作者：
VUORIO, E
影响因子：
4.3
作者：
Miltyk, W;Kama, E;Palka, J
通讯作者：
Palka, J
影响因子：
5.9
作者：
GOODMAN, SI;SOLOMONS, CC;OBRIEN, D
通讯作者：
OBRIEN, D