羧胺三唑乳清酸盐对小鼠胶质瘤相关巨噬细胞系促癌表型的影响  

Effects of carboxyamidotriazole-orotate on oncogenic phenotypes of mouse glioma-associated macrophage cell lines

作　　者：马瑞 杨黎星 陈秋霞 仇佳星 王钰铖 鞠瑞[1] 郭磊[1] MA Rui;YANG Li-xing;CHEN Qiu-xia;QIU Jia-xing;WANG Yu-cheng;JU Rui;GUO Lei(Department of Pharmacology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院药理系,北京100005

出　　处：《基础医学与临床》2022年第5期701-707,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81872897,81672966,82002094)。

摘　　要：目的初步探索羧胺三唑乳清酸盐(CTO)对小鼠胶质瘤细胞系GL261和肿瘤相关巨噬细胞(TAMs)系促癌表型的调控作用及其机制。方法以GL261细胞培养上清诱导骨髓来源巨噬细胞(BMDM)或巨噬细胞系RAW264.7作为研究对象,用qPCR检测TAMs促癌介质mRNA水平;通过Seahorse细胞能量测定方法检测TAMs耗氧速率(OCR);用Western blot检测TAMs中低氧诱导因子-1α(HIF-1α)、低氧诱导因子-2α(HIF-2α)及过氧化物酶体增殖物激活受体(PPAR)共激活因子-1β(PGC-1β)蛋白水平。结果CTO降低TAMs中IL-1β、IL-6、TNF-α等经典活化巨噬细胞(M1)相关介质(P<0.001)及IL-10、Arg-1、TGF-β1等替代性活化巨噬细胞(M2)相关介质的mRNA水平(P<0.001);CTO显著下调TAMs中OCR(P<0.01);CTO下调TAMs中HIF-1α及HIF-2α蛋白水平(P<0.01),这一作用与促进线粒体耗氧的氧化磷酸化解偶联剂碳酰氰-4-三氟甲氧基苯腙(FCCP)作用相反;CTO显著降低TAMs中PGC-1β蛋白水平(P<0.001),这一作用与线粒体呼吸链抑制剂鱼藤酮(RTN)的作用一致。结论CTO抑制氧化磷酸化下调PGC-1β表达,并且使HIF-1α及HIF-2α不稳定,从而影响TAMs中M1及M2表型的促癌介质生成。Objective To explore regulation mechanism of carboxyamidotriazole-orotate(CTO)on oncogenic phenotype of mouse glioma-associated macrophages.Methods Bone marrow-derived macrophages(BMDMs)or RAW264.7 macrophages(tumor associated macrophages,TAMs)were prepared by super-culture of mouse glioma GL261 cells.The mRNA of TAMs oncogenic mediators was detected by qPCR.The oxygen consumption rate(OCR)of TAMs was measured by Seahorse bioenergy method.The protein level of hypoxia-inducible factor-1α(HIF-1α),hypoxia-inducible factor-2α(HIF-2α)and peroxisome proliferator-activated receptor(PPAR)co-activator factor-1β(PGC-1β)in TAMs was detected by Western blot.Results CTO decreased M1-related mediators such as IL-1β,IL-6 and TNF-α(P<0.001)and M2-related mediators such as IL-10,ARG-1 and TGF-β1(P<0.001)in TAMs.CTO significantly down-regulated OCR in TAMs(P<0.01);The protein level of HIF-1αand of HIF-2αin TAMs was down-regulated by CTO(P<0.01),which was contrary to the effect of carboxycyanide-4-trifluormethoxyphenylhydrazone(FCCP),which in turn promoted mitochondrial oxygen consumption.CTO significantly reduced PGC-1βprotein in TAMs(P<0.001),an effect consistent with rotenone(RTN),a mitochondrial respiratory chain inhibitor.ConclusionsA decreased oxidative phosphorylation down-regulates the expression of PGC-1βand destabilizes HIF-1αand HIF-2α,thus affects the production of several M1 and M2 phenotypes of carcinogenic mediators in TAMs,which may be a potential mechanism through which CTO improves the chemotherapy efficacy of glioblastoma.

关 键 词：羧胺三唑乳清酸盐 肿瘤相关巨噬细胞 氧化磷酸化 低氧诱导因子 PPAR共激活因子-1β 

分 类 号：R96[医药卫生—药理学]