MiR-130b调控巨噬细胞极化的分子机制研究  

Mechanism of miR-130b regulating macrophage polarization

作　　者：张亮[1] 赵滢[2] 张天彪[3] 

机构地区：[1]辽宁省肿瘤医院胸外科,辽宁沈阳110042 [2]中国医科大学附属盛京医院普外科,辽宁沈阳110004 [3]中国医科大学生物化学与分子生物学教研室,辽宁沈阳110001

出　　处：《现代肿瘤医学》2015年第6期738-742,共5页Journal of Modern Oncology

基　　金：辽宁省科技厅基金项目(编号:2013225021)

摘　　要：目的:研究miR-130b对巨噬细胞极化的调控作用及机制。方法:选用人白血病单核THP-1细胞,通过PMA使其诱导为Mφ巨噬细胞,然后加入LPS诱导为M1或加入IL-4诱导为M2巨噬细胞;流式细胞仪检测标记物比例;应用Real-time PCR和Western blot方法检测特异性标志物和目的蛋白的mRNA和蛋白表达水平;ELISA法检测特异性分泌因子的表达水平。结果:与Mφ巨噬细胞比较,LPS诱导的巨噬细胞中CD86高表达,TNFα和IL-1β表达和分泌水平均增多,符合M1巨噬细胞特征;IL-4诱导的巨噬细胞中CD206的阳性表达率升高,CCL22、CCL17表达和分泌水平均增多,符合M2巨噬细胞特征;M1细胞中miR-130b表达明显高于M2细胞(P<0.05);miR-130b过表达后的M2细胞中,CCL22 mRNA和蛋白表达减少,IL-1βmRNA和蛋白表达增多,PPAR-γ的蛋白表达降低。结论:MiR-130b可能通过靶向抑制PPAR-γ表达调节巨噬细胞极化。Objective：To study the mechanism of miR-130b regulating macrophage polarization.Methods：Human leukemia mononuclear THP 1 cells was induced to Mφ macrophages by PMA,then add LPS induce to M1 or add IL-4 to for m^2 macrophages.The specific markers were measured using flow cytometry,Real-time PCR and Western blot methods.Secrete factors of macrophages was detected by ELISA method.Results：Compared with Mφ macrophages,CD86 high expression,TNF alpha and IL-1 beta expression and secretion levels were increased when macrophages induced by LPS,display M1 macrophage characteristics.In macrophages induced by IL-4,the positive expression of CD206 rate increased,CCL22,CCL17 expression and secretion levels were increased.MiR-130 b was expressed at a higher level in M1（ P〈0.05）.Overexpression of miR-130 b in m^2 cells decreased CCL22 mRNA and protein level,increased IL-1β mRNA and protein level,reduced the protein expression of PPAR-gamma.Conclusion：MiR-130 b regulates macrophage polarization by targeting PPAR-γ.

关 键 词：miR-130b M1巨噬细胞 M2巨噬细胞 极化 

分 类 号：R730[医药卫生—肿瘤]