a Department of Radiation Oncology , Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University , Nanjing , PR China.
b Department of General Surgery , Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University , Nanjing , PR China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):833-843. doi: 10.1080/21691401.2019.1576711.
We aimed to explore the mediating role of Notch signal in macrophage polarization. This signal was knocked out from macrophages of Lyz2 cre and RBP-J flox mice. Bone marrow-derived macrophages (BMDMs) were isolated and polarized. The expressions of polarization markers in BMDMs 24 h after transfection were detected by qPCR. After Notch knockout, the expressions of M1 markers decreased but those of M2 markers increased significantly. MiR-125a/miR-99b and Spaca6 were highly and lowly expressed upon M1 and M2 polarizations, respectively. The expressions of experimental group were significantly lower than those of control group. Overexpression of miR-125a significantly promoted the expressions of M1 markers, whereas inhibited those of M2 markers. NO release in the culture supernatant of miR-125a overexpression group significantly exceeded that of control group. Transfection with miR-125a inhibitor significantly down-regulated IL-12 expression but up-regulated MR expression in BMDMs. The supernatant secreted by M1 macrophages significantly facilitated BS524 cell apoptosis, which was enhanced after miR-125a overexpression. The TNF-α expression of miR-99b overexpression group increased whereas that of MR decreased significantly. The miR-125a/miR-99b cluster contained an RBP-J specific recognition site in the first intron of initial transcript. The Notch signalling pathway promoted macrophage polarization into M1 phenotype by up-regulating miR-125a/miR-99b expression.
我们旨在探索 Notch 信号在巨噬细胞极化中的介导作用。该信号从 Lyz2 cre 和 RBP-J flox 小鼠的巨噬细胞中被敲除。分离并极化骨髓来源的巨噬细胞（BMDM）。转染后 24 小时，通过 qPCR 检测 BMDM 中极化标志物的表达。Notch 敲除后，M1 标志物的表达减少，但 M2 标志物的表达显著增加。miR-125a/miR-99b 和 Spaca6 在 M1 和 M2 极化时分别高度和低度表达。实验组的表达明显低于对照组。miR-125a 的过表达显著促进了 M1 标志物的表达，而抑制了 M2 标志物的表达。miR-125a 过表达组培养上清液中的 NO 释放明显超过对照组。转染 miR-125a 抑制剂可显著下调 BMDM 中 IL-12 的表达，而上调 MR 的表达。M1 巨噬细胞分泌的上清液可显著促进 BS524 细胞凋亡，而 miR-125a 过表达后这种促进作用增强。miR-99b 过表达组的 TNF-α 表达增加，而 MR 的表达显著降低。miR-125a/miR-99b 簇在初始转录物的第一内含子中包含一个 RBP-J 特异性识别位点。Notch 信号通路通过上调 miR-125a/miR-99b 的表达促进巨噬细胞向 M1 表型极化。
