异泽兰黄素通过PDGFβ/ERK信号通路抑制增生性瘢痕生长的机制探讨

吴志贤 李响 莫自增

[摘要]目的：探討异泽兰黄素（Eupatilin）介导PDGFβ对增生性瘢痕成纤维细胞增殖抑制作用的分子机制。方法：应用组织贴壁法从增生性瘢痕和正常皮肤组织中分离增生性瘢痕和正常皮肤成纤维细胞，Western blot法检测PDGFβ在增生性瘢痕和正常皮肤成纤维细胞中的表达，通过脂质体转染pcDNA3.1-PDGFβ构建PDGFβ过表达增生性瘢痕成纤维细胞，CCK8法检测过表达PDGFβ对异泽兰黄素抑制增生性瘢痕成纤维细胞活力的影响，流式细胞仪检测过表达PDGFβ对异泽兰黄素促进增生性瘢痕成纤维细胞凋亡的影响，Western blot法检测过表达PDGFβ对异泽兰黄素调控细胞内p-ERK、Bcl2和Bax蛋白表达的影响。结果：免疫组化和Western blot结果表明，与正常皮肤组织和成纤维细胞比较，增生性瘢痕组织和成纤维细胞中PDGFβ高表达;CCK8结果显示，与空载对照组（pcDNA31-control）比较，PDGFβ过表达组瘢痕成纤维细胞活力显著升高（P<0.05）;流式细胞术检测结果表明，与对照组pcDNA3.1-control+Eupatilin比较，pcDNA3.1-PDGFβ+Eupatilin组增生性瘢痕成纤维细胞凋亡率显著降低（P<0.05）;Western blot结果显示，PDGFβ过表达能显著促进p-ERK和Bcl2的表达增加，抑制Bax蛋白表达（P<0.05），与对照组相比具有显著统计学差异。结论：异泽兰黄素可抑制增生性瘢痕PDGFβ蛋白的表达，通过抑制PDGFβ/ERK通路诱导细胞凋亡。

[关键词]异泽兰黄素;增生性瘢痕;细胞凋亡;PDGFβ;ERK通路

[中图分类号]R619+.6    [文献标志码]A    [文章编号]1008-6455（2019）07-0044-04

Eupatilin Inhibits Hypertrophic Scar Proliferation Via the PDGFβ/ERK Signaling Pathway

WU Zhi-xian， LI Xiang，MO Zi-zeng，LIANG Jie

（Department of Plastic Surgery，Affiliated Hospital of Guangdong Medical University，Zhanjiang 524001，Guangdong，China）

Abstract： Objective  To investigate the molecular mechanism of Eupatilin-mediated PDGFβ inhibiting the proliferation of hypertrophic scar fibroblasts. Methods  The hypertrophic scars and normal skin fibroblasts were separated from hypertrophic scars and normal skin tissue by tissue adherence. The expression of PDGFβ in hypertrophic scars and normal skin fibroblasts were examined by western blot. The effect of PDGFβ on the viability of hypertrophic scar fibroblasts inhibited by Eupatilin was evaluated by CCK8 assay. The effect of PDGFβ on the apoptosis of hypertrophic scar fibroblasts induced by Eupatilin was measured by flow cytometry analysis. The effect of PDGFβ on the expression of ERK， p-ERK， Bcl2 and Bax were examined by western blot analysis. Results   Immunohistochemistry and western blot results showed that PDGFβ was highly expressed in hypertrophic scar tissue and fibroblasts compared with normal skin tissue and fibroblasts. CCK8 results showed that the activity of scar fibroblasts in the PDGFβ overexpression group was significantly higher compared with the empty control group （pcDNA31-control） （P<0.05）. Flow cytometry results showed that the apoptosis rate of the hypertrophic scar fibroblasts in the pcDNA3.1-PDGFβ+Eupatilin group was significantly lower compared with the control group pcDNA3.1-control+Eupatilin （P<0.05）. Western blot results showed that overexpression of PDGFβ significantly promoted the expression of p-ERK and Bcl2 and inhibited the expression of Bax protein， which was significantly different from the control group （P<0.05）. Conclusion  Eupatilin inhibits the expression of PDGFβ protein in hypertrophic scars and induces apoptosis by inhibiting PDGFβ/ERK pathway.

Key words： eupatilin; hypertrophic scar; apoptosis; PDGFβ; ERK signaling pathway

正常皮肤损伤后，创伤后组织修复异常导致增生性瘢痕的产生。增生性瘢痕不仅伴有瘙痒等并发症，而且严重影响美观，给患者带来巨大压力。增生性瘢痕主要表现为多种纤维化因子的过度表达，胶原蛋白等细胞外基质过度合成和降解减少[1]。……