CTO抑制小鼠胶质瘤细胞原位生长并调节相关小胶质细胞代谢及炎性表型  

CTO inhibits the in situ growth of glioblastoma in mice and regulates the metabolism and inflammatory phenotype of glioma-associated microglia

作　　者：李云帆 邹佳铭 王钰铖 鞠瑞[1] 郭磊[1] LI Yunfan;ZOU Jiaming;WANG Yucheng;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》2025年第4期478-485,共8页Basic and Clinical Medicine

基　　金：呼吸和共病全国重点实验室开放课题基金(2060204);科技创新2030-“脑科学与类脑研究”重大项目2021年度定向委托项目(2021ZD0201100);“人脑组织资源库及地区脑库协作网络平台”课题1(2021ZD0201101)。

摘　　要：目的探究羧胺三唑乳清酸盐(CTO)对胶质瘤相关小胶质细胞(GAM)代谢与炎性反应介质的影响与调节机制。方法通过活体成像检测小鼠体内肿瘤体积,HE染色检测肿瘤占位情况;通过非靶向代谢组学分析检测细胞内三羧酸循环代谢物水平;使用Seahorse细胞能量测定方法检测细胞耗氧率(OCR)和细胞外酸化速率(ECAR);细胞免疫荧光检测细胞缺氧水平;使用qPCR检测细胞中M1型巨噬细胞及M2型巨噬细胞促癌介质mRNA水平;通过Western blot检测缺氧诱导因子-1α(HIF-1α)、程序性死亡受体-配体1(PD-L1)的蛋白质水平。结果CTO抑制小鼠体内肿瘤进展,体外实验中CTO下调GAM的氧化磷酸化水平(P<0.01),改善细胞缺氧,下调HIF-1α(P<0.0001),下调促癌介质iNos、Arg-1、Il-10、Irf4 mRNA的表达(P<0.01),合用乳酸脱氢酶抑制剂司替戊醇(STP)可削弱CTO引起的GAM糖酵解增强和PD-L1表达上调(P<0.01),并下调Arg-1、Il-10的表达(P<0.0001)。结论CTO可下调GAM中多种促癌基因的表达,抑制小鼠体内肿瘤进展;合用乳酸脱氢酶抑制剂可削弱CTO的不利作用并降低GAM促癌介质的转录水平。Objective To investigate the effects and regulatory mechanisms of carboxyamidotriazole orotate(CTO)on the metabolism and inflammatory mediators of glioma-associated microglia(GAM).Methods Tumor volume was regularly monitored by in vivo imaging,and histological examination was performed to detect the extent of tumor infiltration;non-targeted metabolomics analysis was used to detect the level of tricarboxylic acid cycle metabolites in cells;seahorse cell energy measurement method was used to detect the oxygen consumption rate(OCR)and extracellular acidification rate(ECAR)of cells;immunofluorescence was used to detect the degree of hypoxia in cells;quantitative PCR was used to detect the mRNA level of pro-cancer mediators M1/M2 in cells;Western blot was used to detect the protein level of hypoxia-inducible factor-1α(HIF-1α)and programmed death receptor-ligand 1(PD-L1).Results CTO inhibited the tumor progression in mice,and down-regulated the oxidative phosphorylation level and improved cell hypoxia in vitro(P<0.01).It also downregulated the expression of pro-oncogenic mediators iNos,Arg-1,Il-10,and Irf4 in GAM(P<0.01).When combined with lactate dehydrogenase inhibitor stiripentol(STP),CTO-induced enhancement of glycolysis and upregulation of PD-L1 expression in GAM was attenuated(P<0.01),and the expressions of Arg-1 and Il-10 was further downregulated(P<0.0001).Conclusions CTO down-regulates the expression of several oncogenic genes in GAM and inhibits tumor progression in mice.Combined use of lactate dehydrogenase inhibitors can weaken the adverse effect of CTO and reduce the transcriptional level of GAM oncogenic mediators.

关 键 词：肿瘤微环境 羧胺三唑乳清酸盐 胶质瘤相关小胶质细胞 

分 类 号：R96[医药卫生—药理学]