Department of Gerontology, First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Department of Intensive Care Unit, Affiliated Hospital of Yangzhou University, Yangzhou, China.
Oxid Med Cell Longev. 2022 Sep 23;2022:4942519. doi: 10.1155/2022/4942519. eCollection 2022.
The macrophage transformation of inflammatory M1 to anti-inflammatory M2 could be promoted by activating PI3K/AKT signaling pathway. In our previous study, it was found that downregulation of lncRNA260 could ameliorate hypoxic cardiomyocyte injury by regulating IL28RA through the activation of PI3K/AKT signaling pathways. It was suggested that lncRNA260 siRNA could promote the macrophages toward M2 polarization by regulating . In this study, lncRNA260 siRNA was used to observe its effect on the polarization of murine bone marrow-derived macrophages (BMDM) and investigate its related mechanisms. lncRNA 260 specific siRNA were designed and synthesized which were transfected into murine BMDM with liposomes. The experiment was divided into three groups: Hypoxia group, Hypoxia+lncRNA 260-specific siRNA transfection group, and Normoxia group. The CD206-APC/CD11b-FITC or CD206-FITC/CD107b (Mac-3) double positive proportions were used to compare the M2 polarization proportions in the hypoxia process by using the immunofluorescence staining method. The p-AKT, Arg 1, PI3KCG, IL28RAV1, and IL28RAV2 protein expression changes were observed by using the western blot method. Compared with the Normoxia group, the M2 proportions were significantly decreased in the Hypoxia group ( < 0.05). Compared with the hypoxia group, the M2 proportions were significantly increased in the Hypoxia+lncRNA260 siRNA transfection group ( < 0.05). In the Hypoxia group, the ratios of Arg 1/-Actin, p-AKT/-Actin, PI3KCG/-Actin, and IL28RAV1/-Actin were significantly lower than those in the Normoxia group ( < 0.05). After transfection with lncRNA260 siRNA, the ratios of Arg1/-Actin, p-AKT/-Actin, PI3KCG/-Actin, and IL28RAV1/-Actin were significantly higher than those in the Hypoxia group ( < 0.05). Compared with the Normoxia group, the IL28RAV2/-Actin in the Hypoxia group was significantly increased ( < 0.05). After transfection with lncRNA260 siRNA, the ratio of IL28RAV2/-Actin was significantly decreased than that in the Hypoxia group ( < 0.05). lncRNA260 siRNA could promote the M2 polarization of the hypoxia macrophages by reducing the IL28RAV2 alternative splicing variant, which might be related to the activation of the JAK-STAT and PI3K/AKT signaling pathways. It will provide a new strategy for the anti-inflammation, antioxidative stress therapy, and cardiac remodeling after AMI.
炎性 M1 向抗炎 M2 的巨噬细胞转化可以通过激活 PI3K/AKT 信号通路来促进。在我们之前的研究中，发现下调长链非编码 RNA260 通过激活 PI3K/AKT 信号通路可以通过调节 IL28RA 来改善低氧心肌细胞损伤。提示 lncRNA260siRNA 可以通过调节. 促进巨噬细胞向 M2 极化。本研究采用 lncRNA260siRNA 观察其对鼠骨髓来源巨噬细胞（BMDM）极化的影响及其相关机制。设计并合成 lncRNA260 特异性 siRNA，用脂质体转染鼠 BMDM。实验分为三组：缺氧组、缺氧+lncRNA260 特异性 siRNA 转染组和常氧组。采用免疫荧光染色法比较缺氧过程中 CD206-APC/CD11b-FITC 或 CD206-FITC/CD107b（Mac-3）双阳性比例，观察 p-AKT、Arg1、PI3KCG、IL28RAV1 和 IL28RAV2 蛋白表达变化。与常氧组相比，缺氧组 M2 比例明显降低（ < 0.05）。与缺氧组相比，缺氧+lncRNA260siRNA 转染组 M2 比例明显升高（ < 0.05）。在缺氧组中，Arg1/-Actin、p-AKT/-Actin、PI3KCG/-Actin 和 IL28RAV1/-Actin 的比值明显低于常氧组（ < 0.05）。转染 lncRNA260siRNA 后，Arg1/-Actin、p-AKT/-Actin、PI3KCG/-Actin 和 IL28RAV1/-Actin 的比值明显高于缺氧组（ < 0.05）。与常氧组相比，缺氧组 IL28RAV2/-Actin 明显增加（ < 0.05）。转染 lncRNA260siRNA 后，IL28RAV2/-Actin 比值明显低于缺氧组（ < 0.05）。lncRNA260siRNA 可通过降低 IL28RAV2 剪接变体促进缺氧巨噬细胞 M2 极化，可能与 JAK-STAT 和 PI3K/AKT 信号通路的激活有关。为 AMI 后抗炎、抗氧化应激治疗和心脏重构提供了新策略。
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