Department of Dermatology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China.
Evid Based Complement Alternat Med. 2012;2012:956107. doi: 10.1155/2012/956107. Epub 2012 May 28.
Re-epithelialization is a crucial step towards wound healing. The traditional Chinese medicine, Astragalus membranaceus (Fisch) Bge, has been used for hundreds of years for many kinds of ulcerated wounds. Recent research has identified the active compound in this drug as astragaloside IV (AS-IV), but the underlying molecular mechanisms of its therapeutic action on keratinocytes remain poorly understood. In this study, we used an in vitro model of ulcer-like wound processes, lithium chloride (LiCl)-induced cultured mouse keratinocytes, to investigate the effects of AS-IV treatment. The effects on cell proliferation were evaluated by the MTS/PMS colorimetric assay, effects on cell migration were determined by a wound-healing scratch experiment, effects on the cell cycle were analyzed by flow cytometry, and effects on protein expression were analyzed by immunoblotting and immunofluorescence. LiCl strongly inhibited cell proliferation and migration, up-regulated β-catenin expression, and down-regulated proliferating cell nuclear antigen (PCNA) expression. AS-IV treatment attenuat the inhibition of proliferation and migration, significantly reducing the enhanced β-catenin expression, and recovering PCNA and β-tubulin expression. Thus, AS-IV mediates mouse keratinocyte proliferation and migration via regulation of the Wnt signaling pathway. Down-regulating β-catenin to increase keratinocyte migration and proliferation is one mechanism by which AS-IV can promote ulcerated wound healing.
再上皮化是伤口愈合的关键步骤。中药黄芪（Fisch）Bge 已被用于治疗多种溃疡性伤口数百年。最近的研究已经确定了这种药物中的活性化合物是黄芪甲苷 IV（AS-IV），但其对角质形成细胞的治疗作用的潜在分子机制仍知之甚少。在这项研究中，我们使用锂盐（LiCl）诱导的培养小鼠角质形成细胞的溃疡性伤口过程的体外模型来研究 AS-IV 治疗的效果。通过 MTS/PMS 比色法评估细胞增殖的影响，通过划痕实验确定对细胞迁移的影响，通过流式细胞术分析细胞周期，通过免疫印迹和免疫荧光分析蛋白表达。LiCl 强烈抑制细胞增殖和迁移，上调β-连环蛋白表达，并下调增殖细胞核抗原（PCNA）表达。AS-IV 处理可减弱增殖和迁移的抑制作用，显著降低增强的β-连环蛋白表达，并恢复 PCNA 和 β-微管蛋白的表达。因此，AS-IV 通过调节 Wnt 信号通路介导小鼠角质形成细胞的增殖和迁移。下调β-连环蛋白以增加角质形成细胞的迁移和增殖是 AS-IV 促进溃疡性伤口愈合的一种机制。
