天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (7): 611-615.doi: 10.11958/20200169

• 细胞与分子生物学 • 上一篇    下一篇

巨噬细胞极化对心肌成纤维细胞活化的影响

吴惠娟1 ，张盛昔1 ，杨潇2 ，胡因铭1 ，王乐旬1△，郭姣1   

  1. 1 广东省代谢病中西医结合研究中心，广东省代谢性疾病中医药防治重点实验室，粤港澳联合代谢病重点实验室，广东药科大 学中医药研究院（邮编 510006）；2 广州市第一人民医院检验科
  • 收稿日期:2020-01-14 修回日期:2020-04-24 出版日期:2020-07-15 发布日期:2020-07-16
  • 作者简介:吴惠娟（1995），女，硕士在读，主要从事中医药防治糖脂代谢病方面研究

Effects of macrophage polarization on the activation of cardiac fibroblast

WU Hui-juan1 , ZHANG Sheng-xi1 , YANG Xiao2 , HU Yin-ming1 , WANG Le-xun1△, GUO Jiao1   

  1. 1 Guangdong Metabolic Disease Research Center of Integrated Chinese and Western Medicine, Guangdong Key Laboratory of Metabolic Disease Prevention and Treatment of Traditional Chinese Medicine, Joint Laboratory of Guangdong, Hong Kong and Macao on Glycolipid Metabolic Diseases, Institute of Chinese Medicine Sciences, Guangdong Pharmaceutical University, Guangzhou 510006, China; 2 Department of Clinical Laboratory, Guangzhou First People's Hospital
  • Received:2020-01-14 Revised:2020-04-24 Published:2020-07-15 Online:2020-07-16

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摘要： 目的 观察巨噬细胞极化上清对心肌成纤维细胞活化的影响。方法 提取SD大鼠的骨髓细胞和心肌成 纤维细胞。利用巨噬细胞集落刺激因子（M-CSF）处理骨髓细胞后，加入刺激因子：M0（无刺激因子）、M1（100 μg/L脂 多糖+10 μg/L干扰素-γ）、M2（20 μg/L白细胞介素-4）诱导巨噬细胞极化。将极化后的不同型别巨噬细胞及其培养 上清分别与心肌成纤维细胞共培养，分别设空白对照组、M0组、M1组和M2组，通过细胞免疫荧光检测心肌成纤维细 胞中纤维化蛋白的表达水平；实时荧光定量逆转录聚合酶链反应检测巨噬细胞和成纤维细胞特征分子的表达； Western blot检测纤维化相关蛋白及转化生长因子β受体（TGFβR）、血小板衍生生长因子受体（PDGFRs）信号通路活 化情况。结果 经M-CSF及相应刺激因子诱导，成功获得M1和M2型巨噬细胞。细胞共培养结果显示，与M0组相 比，M1组上清培养的心肌成纤维细胞中胶原蛋白1（Col1a1）和Col3a1的mRNA水平以及平滑肌肌动蛋白（α-SMA）表 达水平显著降低（P＜0.05），而M2组上清培养的心肌成纤维细胞中Col1a1和Col3a1的mRNA水平以及α-SMA、结缔 组织生长因子（CCN2）表达水平显著升高（P＜0.05）。M1组上清培养的心肌成纤维细胞中PDGFRβ蛋白磷酸化水平 显著低于 M0 组（P＜0.01），而 M2 组上清培养的心肌成纤维细胞中 PDGFRβ 蛋白磷酸化水平显著高于 M0 组（P＜ 0.05）。结论 M1型巨噬细胞上清能够抑制心肌成纤维细胞活化，而M2型巨噬细胞上清能够激活心肌成纤维细胞。 M1型巨噬细胞抑制纤维化的作用可能与抑制PDGFRβ通路的活化有关。

关键词: 纤维化；心脏；成纤维细胞；巨噬细胞；受体, 血小板源生长因子β

Abstract: Objective To observe the effects of macrophage polarization supernatant on the activation of cardiac fibroblasts. Methods Bone marrow cells and cardiac fibroblasts of SD rats were extracted. Bone marrow cells were induced to M1 and M2 by treating with macrophage colony-stimulating factor (M-CSF), and cells were divided into M0 group (no stimulating factor), M1 group (100 μg/L LPS+10 μg/L INF-γ) and M2 group (20 μg/L IL-4). Different macrophages were co-cultured with cardiac fibroblasts, and different macrophage supernatants were collected to culture with cardiac fibroblasts. Immunofluorescence was performed to examine the fibrotic protein expression in cardiac fibroblasts. The mRNA levels of macrophage-specific molecules, fibrosis-related genes and signaling pathways were tested by real-time PCR. The fibrosis-related proteins and the activation of TGFβR and PDGFRs signal pathways were detected by Western blot assay.Results After treatment with M-CSF and stimulating factors, M1 macrophages and M2 macrophages were induced. Compared with the M0 group, the mRNA levels of Col1a1 and Col3a1 and the protein level of α-SMA were significantly decreased in the cardiac fibroblasts treated by the supernatant of M1 macrophage group (P＜0.05), while the mRNA levels of Col1a1 and Col3a1 and the protein levels of CCN2 and α-SMA were significantly increased in the cardiac fibroblasts treated by the supernatant of M2 macrophage group (P＜0.05).The phosphorylated protein level of PDGFRβ was significantly lower in the cardiac fibroblasts treated by the supernatant of M1 macrophage group than those in the cardiac fibroblasts treated by the supernatant of M0 macrophage group (P＜0.01), while phosphorylated PDGFRβ protein levels were significantly higher in the cardiac fibroblasts treated by the supernatant of M2 macrophage group than those in the cardiac fibroblasts treated by the supernatant of M0 macrophage group (P＜0.05). Conclusion M1 macrophage supernatant can inhibit the activation of cardiac fibroblasts, while M2 macrophage supernatant can activate cardiac fibroblasts. The inhibitory effect of M1 macrophages on fibrosis may be related to restraining the activation of PDGFRβ pathway.

Key words: fibrosis, heart, fibroblasts, macrophages, receptor, platelet-derived growth factor beta