摘要
背景:M2型巨噬细胞具有降低炎症因子及促进组织愈合的功能,因此,如何调节巨噬细胞M2极化是近年来研究的热点,研究发现部分miRNAs具有此功能。目的:探讨miR-378a对Raw264.7巨噬细胞系极化的影响。方法:首先使用脂多糖和干扰素γ协同诱导巨噬细胞M1极化,使用白细胞介素4诱导巨噬细胞M2极化,qRT-PCR检测各型细胞中内源性miR-378a的表达,验证miR-378a是否参与巨噬细胞的极化。通过慢病毒载体将miR-378a转染至巨噬细胞,筛选出miR-378a稳定表达的细胞系,使用脂多糖和干扰素γ协同诱导巨噬细胞M1极化,使用白细胞介素4诱导巨噬细胞M2极化,ELISA检测巨噬细胞培养基上清液中M1/M2极化相关细胞因子水平,qRT-PCR检测M1/M2型巨噬细胞极化特征及相关细胞因子的mRNA表达。结果与结论:①诱导巨噬细胞极化后,各组Raw264.7细胞中内源性miR-378a的表达量均升高;②与未转染组比较,转染miR-378a组诱导巨噬细胞M1极化后促炎性细胞因子诱导型一氧化氮合酶、肿瘤坏死因子α、白细胞介素6、白细胞介素1β表达显著降低(P<0.05),细胞上清液中诱导型一氧化氮合酶、肿瘤坏死因子α、白细胞介素6水平也明显降低(P<0.05);③与未转染组比较,转染miR-378a组诱导巨噬细胞M2极化后CD206、白细胞介素10、精氨酸酶Ⅰ的表达显著升高(P<0.05),细胞上清液中CD206、白细胞介素10水平也明显升高(P<0.05);④结果表明:过表达miR-378a促进巨噬细胞向M2极化且抑制巨噬细胞向M1极化。
BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function.OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line.METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines.RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1βin macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-αand interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant
作者
杨泉
何惠宇
王思凡
吕尚毅
周琦琪
韩祥祯
Yang Quan;He Huiyu;Wang Sifan;Lyu Shangyi;Zhou Qiqi;Han Xiangzhen(Department of Oral Emergency Medicine,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China;Xinjiang Institute of Stomatology,Urumqi 830054,Xinjiang Uygur Autonomous Region,China;Department of Prosthodontics,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第13期2036-2041,共6页
Chinese Journal of Tissue Engineering Research
基金
新疆维吾尔自治区自然科学基金计划青年项目(2021D01C337),项目负责人:韩祥祯~。
