Melatonin-stimulated MSC-derived exosomes improve diabetic wound healing through regulating macrophage M1 and M2 polarization by targeting the PTEN/AKT pathway
Melatonin-stimulated MSC-derived exosomes improve diabetic wound healing through regulating macrophage M1 and M2 polarization by targeting the PTEN/AKT pathway
复制标题
褪黑素刺激的 MSC 衍生的外泌体通过靶向 PTEN/AKT 通路调节巨噬细胞 M1 和 M2 极化，改善糖尿病伤口愈合
DOI：
10.1186/s13287-020-01756-x
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发表时间：
2020-06-29
影响因子：
7.5
通讯作者：
Yang, Lili
中科院分区：
文献类型：
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作者：
Liu, Wei;Yu, Muyu;Yang, Lili
背景糖尿病患者手术后，由于迟发性炎症可能导致伤口愈合中断，从而导致不良后果，目前缺乏有效的措施来解决这一问题。间充质干细胞（MSC）来源的外泌体（Exo）已被证明是通过抗炎作用促进糖尿病伤口愈合的合适候选物。在这项研究中，我们调查是否褪黑激素（MT）预处理的MSC衍生的exosomes（MT-Exo）可以发挥上级效果对糖尿病伤口愈合，我们试图阐明其潜在的mechanism.MethodsFor的抗炎作用的评价MT-Exo，在体外和体内研究进行。体外研究采用ELISA法检测IL-1 β、TNF-α、IL-10等炎症相关因子的分泌水平，qRT-PCR法检测IL-1 β、TNF-α、IL-10、Arg-1、iNOS等相关基因的表达，Western blotting法检测PTEN、AKT、p-AKT的表达。在体内研究方面，建立了气囊模型和链脲佐菌素（STZ）处理的糖尿病创面模型，通过流式细胞术、光学成像、H&E染色、Masson三色染色、免疫组织化学染色、免疫荧光和qRT-PCR等方法评价MT-Exo对糖尿病创面愈合的影响（α-SMA，结果MT-Exo能显著抑制促炎因子IL-1β和TNF-α的表达。与PBS组、LPS组和Exo组相比，IL-1β、TNF-α和iNOS的相对基因表达量降低，而抗炎因子IL-10的相对基因表达量沿着IL-10和Arg-1的相对基因表达量增加。这一作用是通过上调PTEN表达和抑制AKT磷酸化而增加M2/M1极化比值介导的。结论MT-Exo可通过激活PTEN/AKT信号通路，增加M2/M1极化比值，抑制炎症反应，促进糖尿病创面愈合。MT预处理被证明是治疗糖尿病伤口愈合的一种有前途的方法。图摘要：MT-Exo通过调节M1和M2巨噬细胞极化促进糖尿病伤口愈合。
BackgroundAfter surgery, wound recovery in diabetic patients may be disrupted due to delayed inflammation, which can lead to undesired consequences, and there is currently a lack of effective measures to address this issue. Mesenchymal stem cell (MSC)-derived exosomes (Exo) have been proven to be appropriate candidates for diabetic wound healing through the anti-inflammatory effects. In this study, we investigated whether melatonin (MT)-pretreated MSCs-derived exosomes (MT-Exo) could exert superior effects on diabetic wound healing, and we attempted to elucidate the underlying mechanism.MethodsFor the evaluation of the anti-inflammatory effect of MT-Exo, in vitro and in vivo studies were performed. For in vitro research, we detected the secreted levels of inflammation-related factors, such as IL-1β, TNF-α and IL-10 via ELISA and the relative gene expression of the IL-1β, TNF-α, IL-10, Arg-1 and iNOS via qRT-PCR and investigated the expression of PTEN, AKT and p-AKT by Western blotting. For in vivo study, we established air pouch model and streptozotocin (STZ)-treated diabetic wound model, and evaluated the effect of MT-Exo by flow cytometry, optical imaging, H&E staining, Masson trichrome staining, immunohistochemical staining, immunofluorescence, and qRT-PCR (α-SMA, collagen I and III).ResultsMT-Exo significantly suppressed the pro-inflammatory factors IL-1β and TNF-α and reduced the relative gene expression of IL-1β, TNF-α and iNOS, while promoting the anti-inflammatory factor IL-10 along with increasing the relative expression of IL-10 and Arg-1, compared with that of the PBS, LPS and the Exo groups in vitro. This effect was mediated by the increased ratio of M2 polarization to M1 polarization through upregulating the expression of PTEN and inhibiting the phosphorylation of AKT. Similarly, MT-Exo significantly promoted the healing of diabetic wounds by inhibiting inflammation, thereby further facilitating angiogenesis and collagen synthesis in vivo.ConclusionsMT-Exo could promote diabetic wound healing by suppressing the inflammatory response, which was achieved by increasing the ratio of M2 polarization to M1 polarization through activating the PTEN/AKT signalling pathway, and the pretreatment of MT was proved to be a promising method for treating diabetic wound healing.Graphical abstract: MT-Exo promotes diabetic wound healing by regulating M1 and M2 macrophage polarization.