摘要
目的:研究绿原酸(CGA)对人肝癌细胞HepG2诱导的肿瘤相关巨噬细胞(TAM)极化的影响。方法:人单核巨噬细胞THP-1经100 mmol/L佛波酯(PMA)刺激48 h诱导分化为M0巨噬细胞,与HepG2细胞共培养,并用不同浓度CGA干预。倒置显微镜下观察细胞形态学变化,CCK-8法检测细胞活性,实时荧光定量PCR(RT-qPCR)法检测TAM相关标志物CD206、白细胞介素(IL)-10、IL-6和环氧化合酶2(COX-2)mRNA表达水平。结果:与HepG2共培养后贴壁M0巨噬细胞形态由扁平圆形转变为不规则圆形,并有伪足伸出和梭形细胞形成,共培养48 h时细胞活性最高(P<0.05)。CGA处理48 h后,在25~100μg/mL浓度范围内随着CGA浓度增加,TAM细胞活性增高(P<0.05)。50μg/mL、100μg/mL CGA处理TAM 48 h后,不规则圆形带伪足细胞和梭形细胞数量增多,M1型巨噬细胞相关标志物IL-6、COX-2表达上调,M2型巨噬细胞相关标志物IL-10、CD163表达下调(P<0.05),且100μg/mL CGA较50μg/mL干预作用更明显。结论:HepG2肝癌细胞能诱导巨噬细胞极化为M2样TAM,CGA干预后TAM向M1样极化转变。
Objective:To investigate the effects of chlorogenic acid(CGA)on human hepatocellular carcinoma cells HepG2-induced tumor-associated macrophage(TAM)polarization.Methods:Human mononuclear macrophage THP-1 was induced to differentiate into M0 macrophages by 100 mmol/L phorbol myristate acetate(PMA)stimulation for 48 h.The cells were co-cultured with HepG2 cells and intervened with different concentrations of CGA.Cell morphological changes were observed under inverted microscope,cell activity was detected by CCK-8 assay,and mRNA expression levels of TAM-related markers CD206,interleukin(IL)-10,IL-6 and cyclooxygenase 2(COX-2)were detected by real-time fluorescence quantitative PCR(RT-qPCR).Results:After co-culture with HepG2,the morphology of adnexal M0 macrophages changed from flattened round to irregular round with pseudopod protrusion and spindle cell formation,and the highest cell activity was observed at 48 h of co-culture(P<0.05).After 48 h of CGA treatment,TAM cell activity increased with increasing CGA concentration in the concentration range of 25-100μg/mL(P<0.05).Treatment of 50μg/mL and 100μg/mL CGA on TAM for 48 h increased the number of irregularly rounded cells with pseudopods and spindle cells and upregulated the expressions of M1 macrophage-related markers IL-6 and COX-2 and downregulated the expressions of M2 macrophagerelated markers IL-10 and CD163(P<0.05),and 100μg/mL CGA had a more pronounced intervening effect than 50μg/mL.Conclusion:HepG2 hepatocellular carcinoma cells could induce macrophage polarization to M2-like TAM,and TAM shifts to M1-like polarization after CGA intervention.
作者
张潇剑
蓝利
阙婷
唐立博
杨子叶
唐深
罗小玲
Zhang Xiaojian;Lan Li;Que Ting;Tang Libo;Yang Ziye;Tang Shen;Luo Xiaoling(School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021,China;Key Laboratory of Basic Medical Research in Universities in Guangxi,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2022年第4期552-556,共5页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.81860504)
广西自然科学基金资助项目(No.2021GXNSFAA196019)。
关键词
绿原酸
肿瘤相关巨噬细胞
HEPG2肝癌细胞
极化
chlorogenic acid
tumor-associated macrophages
HepG2 hepatocellular carcinoma cells
polarization