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裸花紫珠调控M2型巨噬细胞极化对糖尿病足溃疡大鼠伤口愈合及PI3K/AKT/mTOR通路的影响

Effect of Callicarpa nudiflora on wound healing and PI3K/AKT/mTOR pathway by regulating M2 macrophage polarization in rats with diabetic foot ulcer

    摘要
  • 摘要:
    目的 分析裸花紫珠调控M2型巨噬细胞极化对糖尿病足溃疡大鼠伤口愈合及磷脂酰肌醇-3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)通路的影响。
    方法 选取40只大鼠, 将其中10只作为对照组(生理盐水外敷)，剩余30只用于建立糖尿病足溃疡大鼠模型。最终建模成功27只大鼠，并分为模型组(生理盐水外敷)、裸花紫珠组(裸花紫珠外敷)、裸花紫珠联合抑制剂组(裸花紫珠外敷联合M2型巨噬细胞标志蛋白CD206抑制剂)，每组9只。比较各组大鼠创面愈合率，碱性成纤维生长因子(bFGF)、血管内皮生长因子(VEGF)水平，诱导型一氧化氮合酶(iNOS)、CD16/32、CD206、PI3K、AKT、mTOR蛋白相对表达量, PI3K、AKT、mTOR的mRNA表达量。
    结果 干预后第3、7、14天，与对照组大鼠相比，各组大鼠创面愈合率、VEGF、bFGF、CD206相对表达量以及PI3K、AKT、mTOR的mRNA表达量和蛋白相对表达量均降低, iNOS、CD16/32相对表达量升高，差异有统计学意义(P < 0.05); 与模型组大鼠相比，裸花紫珠组、裸花紫珠联合抑制剂组大鼠创面愈合率、VEGF、bFGF、CD206相对表达量以及PI3K、AKT、mTOR的mRNA表达量和蛋白相对表达量均升高, iNOS、CD16/32相对表达量降低，差异有统计学意义(P < 0.05); 与裸花紫珠组大鼠相比，裸花紫珠联合抑制剂组大鼠创面愈合率、VEGF、bFGF、CD206相对表达量以及PI3K、AKT、mTOR的mRNA表达量和蛋白相对表达量均降低, iNOS、CD16/32相对表达量升高，差异有统计学意义(P < 0.05)。
    结论 裸花紫珠可促进M2型巨噬细胞极化，提高糖尿病足溃疡大鼠伤口愈合率以及VEGF、bFGF水平，激活PI3K/AKT/mTOR信号通路。

     

    Abstract:
    Objective To analyze the effect of Callicarpa nudiflora on wound healing and phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway by regulating M2 macrophage polarization in rats with diabetic foot ulcer.
    Methods A total of 40 rats were selected, with 10 rats in control group (topical application of saline), and the remaining 30 were used to establish rat model of diabetic foot ulcer. Ultimately, 27 rats were successfully modeled and divided into model group (topical application of saline), Callicarpa nudiflora group (topical application of Callicarpa nudiflora), and Callicarpa nudiflora combined with inhibitor group (topical application of Callicarpa nudiflora combined with CD206 inhibitor, a marker protein of M2 macrophages), with 9 rats in each group. The wound healing rate, levels of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), relative expression levels of inducible nitric oxide synthase (iNOS), CD16/32, CD206, PI3K, AKT and mTOR as well as mRNA expression levels of PI3K, AKT and mTOR were compared among groups.
    Results On the 3rd, 7th and 14th days after intervention, compared with the control group, the wound healing rate, VEGF, bFGF, relative expression level of CD206, relative expression of PI3K, AKT and mTOR at mRNA and protein levels were significantly decreased while the relative expression levels of iNOS and CD16/32 were significantly increased in all other groups (P < 0.05); compared with the model group, the Callicarpa nudiflora group and the Callicarpa nudiflora combined with inhibitor group showed significant increased wound healing rate, VEGF, bFGF, relative expression level of CD206, and relative expression of PI3K, AKT and mTOR at mRNA and protein levels, and significant decreased relative expression levels of iNOS and CD16/32 (P < 0.05); compared with the Callicarpa nudiflora group, the Callicarpa nudiflora combined with inhibitor group showed significant decreased wound healing rate, VEGF, bFGF, relative expression level of CD206, and relative expression of PI3K, AKT and mTOR at mRNA and protein levels, and significant increased relative expression levels of iNOS and CD16/32 (P < 0.05).
    Conclusion Callicarpa nudiflora can promote M2 macrophage polarization, increase wound healing rate and levels of VEGF and bFGF in rats with diabetic foot ulcer, and activate the PI3K/AKT/mTOR signaling pathway.

     

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