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[富血小板血浆通过调节AMPK信号通路诱导M2巨噬细胞极化]

[Platelet-rich Plasma Induces M2 Macrophage Polarization via Regulating AMPK Singling Pathway].

作者信息

Shi Lin-Ying, Li Yan-Hui, Xu Jing-Jing, Zhang Yu, Xie Ting-Ting, Xu Yu-Bing, Shan Gui-Qiu, Zhou Mou

机构信息

Department of Blood Transfusion, General Hospital of Southern Theatre Command of PLA, Guangzhou 510010, Guangdong Province, China.

Department of Blood Transfusion, General Hospital of Southern Theatre Command of PLA, Guangzhou 510010, Guangdong Province, China.E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Oct;31(5):1486-1491. doi: 10.19746/j.cnki.issn.1009-2137.2023.05.037.

Abstract

OBJECTIVE

To investigate the role of platelet-rich plasma (PRP) in inducing the M2 macrophage polarization via regulating AMPK singling pathway.

METHODS

The expressions of M1 marker CD11c and M2 marker CD206 in macrophages of blank control group, LPS group, LPS+PRP group, and LPS+PRP+Compound C group were detected by flow cytometry. Western blot was used to observe the effects of PRP on the expression of AMPK-mTOR signaling pathway-related proteins at different times (12 h, 18 h and 24 h) after LPS treatment. RNA interference technology was used to silence the expression of AMPK in macrophages, and the expression of TGF-β protein was subsequently examined by Western blot.

RESULTS

LPS significantly reduced the expression of CD206 and increased the expression of CD11c ( <0.05). After the addition of PRP, the expression of CD206 was significantly increased ( <0.05), while the expression of CD11c was significantly decreased ( <0.05). Compared with LPS group, PRP treatment significantly increased the expressions of p-AMPK and p-ULK1 proteins at 12 h, 18 h and 24 h, while significantly decreased the expression of p-mTOR protein ( <0.05). After the addition of AMPK inhibitor Compound C, the expression of CD206 was significantly reduced ( <0.05) and the expression of CD11c was significantly increased compared with LPS+PRP group ( <0.05). After silencing the expression of AMPK in macrophages, the promotion effect of PRP on TGF-β was significantly reduced ( <0.05).

CONCLUSION

PRP can stimulate the transformation of macrophages to M2 type via AMPK signalling pathway.

摘要

目的

探讨富血小板血浆(PRP)通过调节AMPK信号通路诱导M2型巨噬细胞极化的作用。

方法

采用流式细胞术检测空白对照组、LPS组、LPS+PRP组和LPS+PRP+Compound C组巨噬细胞中M1标志物CD11c和M2标志物CD206的表达。采用蛋白质免疫印迹法观察PRP对LPS处理后不同时间点(12 h、18 h和24 h)AMPK-mTOR信号通路相关蛋白表达的影响。采用RNA干扰技术沉默巨噬细胞中AMPK的表达,随后通过蛋白质免疫印迹法检测TGF-β蛋白的表达。

结果

LPS显著降低CD206的表达,增加CD11c的表达(<0.05)。加入PRP后,CD206的表达显著增加(<0.05),而CD11c的表达显著降低(<0.05)。与LPS组相比,PRP处理在12 h、18 h和24 h显著增加p-AMPK和p-ULK1蛋白的表达,同时显著降低p-mTOR蛋白的表达(<0.05)。加入AMPK抑制剂Compound C后,与LPS+PRP组相比,CD206的表达显著降低(<0.05),CD11c的表达显著增加(<0.05)。沉默巨噬细胞中AMPK的表达后,PRP对TGF-β的促进作用显著降低(<0.05)。

结论

PRP可通过AMPK信号通路刺激巨噬细胞向M2型转化。

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