Periplocin promotes wound healing through the activation of Src/ERK and PI3K/Akt pathways mediated by Na/K-ATPase
Periplocin promotes wound healing through the activation of Src/ERK and PI3K/Akt pathways mediated by Na/K-ATPase
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Periplocin 通过激活 Na/K-ATPase 介导的 Src/ERK 和 PI3K/Akt 途径促进伤口愈合
DOI：
10.1016/j.phymed.2018.12.015
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发表时间：
2019-04-01
期刊：
影响因子：
7.9
通讯作者：
Liu, Wenyuan
中科院分区：
文献类型：
--
作者：
Chen, Lei;Jiang, Pan;Liu, Wenyuan
背景：香附(Periploca forrestii，PF)过去主要用于治疗关节炎和创伤。我们先前已经证明，从PF中分离出的富含强心类固醇的一部分具有促进伤口愈合的潜力。然而，促进创面愈合的确切物质基础和作用机制仍不清楚。Periplocin(PP)是PF中强心类固醇的最高含量。本研究旨在系统地评价香附素体内外对创面愈合的影响。材料与方法：采用四甲基偶氮唑盐比色法和EDU比色法检测L929细胞增殖情况。用划痕法和Transwell法检测细胞迁移情况。用Sircol型胶原蛋白检测试剂盒检测可溶性胶原的总量。用切除大鼠模型进行体内创面愈合活性评价。用苏木精-伊红染色观察伤后第9天皮肤的大体形态变化，用Masson‘s三色染色观察胶原沉积。Western印迹法检测Perplocin处理后，Na/K-ATPase介导的Src/ERK和PI3K/Akt信号通路的改变。结果：香附红素可显著促进成纤维细胞L929的增殖、迁移和胶原生成，其作用依赖于Na/K-ATPase介导的Src/ERK和PI3K/Akt通路的激活，从而促进创面愈合。确实，用Src特异性抑制剂抑制Na/K-ATPase/Src复合体受体或下调Na/K-ATPase的表达将取消随后的Src/ERK和PI3K/Akt通路的激活，并减弱周围素对创面愈合的有利作用。此外，在大鼠切除创面模型中，创面愈合活性也得到了证实，创面闭合率、再上皮化率、肉芽组织形成和胶原沉积的增加都证明了这一点。结论：总而言之，我们为创伤的传统治疗奠定了基础，表明香附和香附是治疗慢性创面的一种有前途的候选药物。
Background: Periploca forrestii(PF) is mainly utilized for treatment of arthritis and traumatic injury historically. We had previously demonstrated that a fraction rich in cardiotonic steroids isolated from PF had the potential to facilitate wound healing. However, the exact material basis and mechanism of action responsible for wound healing is still unclear. Periplocin(PP) is the highest level of cardiotonic steroid included in PF. The present study aims to evaluate the efficacy of periplocin on wound healing systematically in vitro and in vivo.Materials and methods: The L929 proliferation was determined by both MTT and EdU assay. Cell migration was tested by both scratch and transwell assay. The total amount of soluble collagen was assessed using a Sircol Collagen Assay Kit. The wound healing activity was evaluated in vivo using the excision rat models. Histopathology of the wounded skin on day 9 was studied via hematoxylin and eosin staining (HE) for general morphological observations and masson's trichrome staining for collagen deposition, respectively. The alteration in Src/ERK and PI3K/Akt pathways mediated by Na/K-ATPase was determined by western blot after the treatment with periplocin. The interaction between Na/K-ATPase and Src was tested by immunoprecipitation and immunostaining analysis.Results: The results revealed that periplocin could significantly boost proliferation, migration and stimulate collagen production in fibroblast L929 cells, which is dependent on activation of Src/ERK and PI3K/Akt pathways mediated by Na/K-ATPase, and thus promoting wound healing. Indeed, inhibition of Na/K-ATPase/Src complex receptor by Src specific inhibitor or knocking down the Na/K-ATPase expression would abolish the subsequent activation of Src/ERK and PI3K/Akt pathways and attenuate periplocin-induced beneficial effects on wound healing. Additionally, the wound healing activity is also confirmed in a rat excisional wound model as evidenced by increased rate of wound closure, reepithelization, formation of granulation tissue and collagen accumulation.Conclusions: Collectively, we lay the rationale for traditional usage for traumatic injury, suggesting that periplocin and periploca forrestii is a promising candidate for management of chronic wounds.