[关键词]
[摘要]
目的 考察王不留行黄酮苷促创伤愈合作用,并探讨其作用机制。方法 构建SD大鼠皮肤开放性创伤模型,创伤部位分别涂抹0.02 g空白软膏剂基质(模型组)、0.02 g含0.1%王不留行黄酮苷的软膏剂(王不留行黄酮苷组)、0.02 g美宝润湿烧伤膏(阳性药)。观察创伤愈合速率,并取创伤部位皮肤制作石蜡切片,通过HE染色进行组织病理学评估,通过免疫组化染色观察增殖细胞核抗原(PCNA)、p-碱性成纤维细胞生长因子受体(p-bFGFR)、p-血管内皮细胞生长因子受体(p-VEGFR)、CD31、p-Akt、p-Erk表达,Western blotting法分析Erk和Akt蛋白的磷酸化水平、bFGFR磷酸化水平。结果 与模型组比较,创伤后3、6、9 d,王不留行黄酮苷组显著促进开放性创伤愈合(P<0.05、0.01);随着时间的延长,与模型组比较,王不留行黄酮苷组中成纤维细胞和内皮细胞大量增殖,炎症细胞增殖减少,微血管密度显著增加(P<0.01);免疫组化及Western blotting结果显示,与模型组比较,王不留行黄酮苷组内皮细胞膜受体中bFGFR的磷酸化程度明显升高(P<0.05、0.01),p-VEGFR磷酸化程度无明显变化,PI3K/Akt与MAPK/Erk信号通路的节点蛋白Akt和Erk磷酸化程度均明显升高(P<0.05)。结论 王不留行黄酮苷促进开放性创伤愈合,机制可能与激活bFGFR及其下游MAPK/Erk和PI3K/Akt信号通路相关。
[Key word]
[Abstract]
Objective To investigate the effect of vaccarin on wound healing and to explore the mechanism of vaccarin promoting wound healing. Methods The open wound model of SD rat skin was constructed, and the wound sites were smeared with 0.02 g blank cream matrix (model group), 0.02 g containing 0.1% vaccarin (vaccarin group) or 0.02 g MEBO wetting burn ointment (positive drug). The rate of wound healing was observed and paraffin sections were made from the skin of the wound. Histopathological evaluation was performed by HE staining. Proliferating cell nuclear antigen (PCNA), p-Akt, p-Erk P-basic fibroblast growth factor receptor (p-bFGFR), p-vascular endothelial cell growth factor receptor (p-VEGFR), and CD31 were observed by immunohistochemical staining. Western blotting method were used to analyze the phosphorylation level of Erk, Akt, and bFGFR protein. Results Compared with model group, 3, 6, and 9 d after trauma, vaccarin significantly promoted the healing of open wound healing (P<0.05, 0.01). Compared with model group, the fibroblasts and endothelial cells in vaccarin group were proliferating, the proliferation of the inflammatory cells decreased and the microvessel density increased significantly (P<0.01); The results of immunohistochemistry and Western blotting showed that the degree of phosphorylation of bFGFR in the endothelial cell membrane receptor of vaccarin group was significantly higher than that in model group (P<0.05, 0.01), and the degree of p-VEGFR phosphorylation was not significantly changed, and the degree of Akt and Erk phosphorylation of PI3K/Akt and MAPK/Erk signaling pathway were significantly increased (P<0.05). Conclusion Vaccarin could promote open wound healing, and may be related to activation of bFGFR and its downstream MAPK/Erk and PI3K/Akt signaling pathways.
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[基金项目]
中央高校基本科研业务费专项(JUSRP51412B)