School of Basic Medical Sciences, Ningxia Medical University, Ningxia 750004, China.
Postgraduate Training Base in Shanghai Gongli Hospital, Ningxia Medical University, Shanghai 200135, China.
Biomed Res Int. 2023 Jan 6;2023:1277258. doi: 10.1155/2023/1277258. eCollection 2023.
In this study, the expression of Cripto-1 and the role of macrophage polarization in immune response after allogeneic transfusion were analyzed by constructing a mouse model of allogeneic transfusion. In order to analyze the effects of miR-449a on the PI3K/AKT/NF-B signaling pathway and the expression of downstream related regulatory factors under normal and abnormal conditions, we adopt in vitro and in vivo experiments separately. The molecular mechanism of PI3K/AKT/NF-B signaling pathway was analyzed by blocking or activating gene expression and western blotting. Experiment in vitro has confirmed that inhibition of miR-449a increased the protein expression of Cripto-1. In vivo experiments confirmed that allogeneic transfusion reduced the expression of Cripto-1, which further inhibited NF-B signaling pathway through AKT/PI3K phosphorylation, regulated macrophage polarization, inhibited M1 polarization of macrophages, promoted M2 polarization, and thus affected immune response of the body.
在这项研究中,通过构建同种异体输血小鼠模型,分析了 Cripto-1 的表达和巨噬细胞极化在同种异体输血后免疫反应中的作用。为了分析 miR-449a 在正常和异常条件下对 PI3K/AKT/NF-B 信号通路和下游相关调节因子表达的影响,我们分别进行了体外和体内实验。通过阻断或激活基因表达和 Western blot 分析了 PI3K/AKT/NF-B 信号通路的分子机制。体外实验证实,抑制 miR-449a 增加了 Cripto-1 的蛋白表达。体内实验证实,同种异体输血降低了 Cripto-1 的表达,进一步通过 AKT/PI3K 磷酸化抑制 NF-B 信号通路,调节巨噬细胞极化,抑制巨噬细胞 M1 极化,促进 M2 极化,从而影响机体的免疫反应。