Induction of PDGF-B in TCA-treated epidermal keratinocytes
Induction of PDGF-B in TCA-treated epidermal keratinocytes
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DOI：
10.1007/s00403-007-0781-6
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发表时间：
2007-08
影响因子：
3
通讯作者：
Nozomi Yonei;N. Kanazawa;T. Ohtani;F. Furukawa;Yuki Yamamoto
中科院分区：
文献类型：
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作者：
Nozomi Yonei;N. Kanazawa;T. Ohtani;F. Furukawa;Yuki Yamamoto
三氯乙酸（TCA）是使用最广泛的脱皮剂之一，它能诱导整个表皮完全坏死，随后表皮和乳头状真皮层基质重建。TCA的细胞毒性作用，如抑制角质形成细胞和成纤维细胞的增殖以及成纤维细胞的蛋白质合成，已经有报道。然而，TCA脱皮的整个生物学机制尚未确定。研究了TCA处理对表皮细胞诱导生长因子和细胞因子产生的假设激活作用，并在时间过程和应用TCA浓度方面与其细胞毒性作用进行了比较。经过不同时间的TCA孵育，用MTT法和染料排除法研究Pam212小鼠角质形成细胞的活力，用逆转录聚合酶链反应（RT-PCR）研究生长因子和细胞因子的产生。应用RT-PCR和免疫组织化学分别检测TCA应用后人皮肤标本中血小板衍生生长因子(PDGF)-B mRNA表达和蛋白生成的变化。与TCA孵育显示细胞毒性并诱导Pam212细胞死亡，这取决于孵育时间和TCA浓度。此外，经短期TCA治疗后，Pam212细胞中均检测到PDGF-B、肿瘤生长因子(TGF)-α、TGF- β1和血管内皮生长因子的表达，这些生长因子是角质形成细胞在伤口愈合过程中分泌的生长因子。炎症细胞因子如白细胞介素(IL)-1和IL-10的表达也被诱导。相反，在tca处理的NIH-3T3成纤维细胞中，观察到只有角质细胞生长因子（据报道是由成纤维细胞分泌的）的上调，以及类似的细胞毒性作用。在人皮肤中，应用TCA后PDGF-B mRNA表达显著上调，然后立即下调。施用TCA后，角质形成细胞细胞质中的免疫反应性PDGF-B在整个表皮中均可检测到，在mRNA表达达到峰值后达到峰值，在表皮完全坏死后24 h内显著下降。tca处理的表皮在发生完全坏死之前是生长因子（包括PDGF-B）的主要来源。这种作用可能有助于促进再上皮化和真皮再生，而不会产生伤口收缩和疤痕。
Trichloroacetic acid (TCA) is one of the most widely used peeling agents, and induces full necrosis of the whole epidermis, followed by reconstitution of the epidermis and the matrix of the papillary dermis. The cytotoxic effects of TCA, such as suppressing proliferation of keratinocytes and fibroblasts and protein synthesis by fibroblasts, have already been reported. However, the entire biological mechanism responsible for TCA peeling has yet to be determined. Hypothetical activation effects of TCA treatment on epidermal cells to induce production of growth factors and cytokines are examined, and are compared with its cytotoxic effects in terms of time course and applied TCA concentrations. After various periods of incubation with TCA, viability of Pam212 murine keratinocytes was investigated with MTT assay and dye exclusion assay, and production of growth factors and cytokines with reverse transcription-polymerase chain reaction (RT-PCR). Changes in platelet-derived growth factor (PDGF)-B mRNA expression and protein production in the human skin specimens after TCA application were then examined by RT-PCR and immunohistochemistry, respectively. Incubation with TCA showed cytotoxicity and induced death of Pam212 cells, depending on the incubation period and the TCA concentration. In addition, expressions of PDGF-B, tumor growth factor (TGF)-α, TGF- β1 and vascular endothelial growth factor, which are the growth factors reportedly secreted from keratinocytes during wound healing, were all detected in Pam212 cells after short-term treatment with TCA. Expressions of inflammatory cytokines such as interleukin (IL)-1 and IL-10 were also induced. In TCA-treated NIH-3T3 fibroblasts, in contrast, observed was upregulation of only keratinocyte growth factor, which is reportedly secreted from fibroblasts, as well as the similar cytotoxic effect. In human skin, PDGF-B mRNA expression became significantly upregulated after TCA application, and then immediately downregulated. Immunoreactive PDGF-B in the cytoplasm of keratinocytes became detectable throughout the epidermis after TCA application, reached maximum after the peak of mRNA expression, and then declined significantly over 24 h when the epidermis became completely necrotic. The TCA-treated epidermis acts as a major source of growth factors, including PDGF-B, before undergoing full necrosis. This effect might contribute to a promotion of re-epithelialization and dermal regeneration without wound contraction and scarring.