1,8-桉叶油素对ox-LDL诱导的巨噬细胞源性泡沫细胞形成的影响及机制  

Effects of 1,8-Cineole on the formation of ox-LDL induced macrophage derived foam cells and its mechanism

作　　者：祝知行 向军 陈星月 陶俊鹭 潘桂先 张光琼 ZHU Zhixing;XIANG Jun;CHEN Xingyue;TAO Junlu;PAN Guixian;ZHANG Guangqiong(Ethnic Medicine and Traditional Chinese Medicine Experimental Teaching Center,Guizhou Medical University,Guiyang 550001,Guizhou,China)

机构地区：[1]贵州医科大学民族药与中药实验教学中心,贵州贵阳550001

出　　处：《贵州医科大学学报》2025年第2期212-219,239,共9页Journal of Guizhou Medical University

基　　金：贵州省卫生健康委员会科学技术基金项目(gzwkj2024-243)。

摘　　要：目的探讨1,8-桉叶油素(1,8-Cineole)对氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的巨噬细胞源性泡沫细胞形成的影响及机制。方法取对数生长期的人单核细胞THP-1,使用100.00μg/L佛波酯(phorbol 12-myristate 13-acetate,PMA)诱导分化成THP-1源性巨噬细胞,将THP-1源性巨噬细胞分为Control组(同等体积的培养基)和各浓度ox-LDL组(0.08 mg/L、0.80 mg/L、8.00 mg/L及80.00 mg/L),采用四甲基偶氮唑盐(methylthiazolyldiphenyl-tetrazolium bromide,MTT)实验检测ox-LDL对THP-1源性巨噬细胞的毒性;将THP-1源性巨噬细胞分为Control组和各浓度1,8-Cineole组(1.00μmol/L、5.00μmol/L、25.00μmol/L、125.00μmol/L及625.00μmol/L),采用MTT实验检测1,8-Cineole对THP-1源性巨噬细胞活性的影响;以浓度为80.00 mg/L ox-LDL诱导THP-1源性巨噬细胞分化为泡沫细胞,将泡沫细胞分为Control组、ox-LDL组(80.00 mg/L ox-LDL)及1,8-Cineole组[ox-LDL 80.00 mg/L+各浓度1,8-Cineole(1.00μmol/L、5.00μmol/L、25.00μmol/L、125.00μmol/L及625.00μmol/L)],采用MTT实验检测1,8-Cineole对ox-LDL诱导THP-1泡沫细胞活力的影响;80.00 mg/L ox-LDL诱导THP-1源性巨噬细胞分化为泡沫细胞模型,分为Control组(同等体积的培养基)、ox-LDL组(80.00 mg/L ox-LDL)、低剂量1,8-Cineole组(80.00 mg/L ox-LDL和1.00μmol/L 1,8-Cineole)及高剂量1,8-Cineole组(80.00 mg/L ox-LDL和5.00μmol/L 1,8-Cineole),采用油红O染色实验检测脂质的积累和泡沫细胞的形成情况,尼罗红染色实验检测细胞脂质摄取情况,蛋白免疫印迹(Western blot)法分析巨噬细胞中A1类清道夫受体(A1 scavenger receptor,SR-A1)蛋白的表达,实时荧光定量PCR(real time fluorescence quantitative PCR,qRT-PCR)检测SR-A1信使RNA(messenge RNA,mRNA)的表达。结果与Control组比较,0.08~80.00 mg/L ox-LDL组和1.00~125.00μmol/L 1,8-Cineole组对THP-1巨噬细胞均无毒性(P>0.05);与ox-LDL组比较,625.00μmol/L 1,8-Cineole组细胞活力降低(P<0.05);与Control组比较,oObjective To investigate the effects and mechanism of 1,8-cineole on the formation of macrophage derived foam cells induced by oxidized low density lipoprotein(ox-LDL).Methods Human monocytes THP-1 in logarithmic growth phase were taken and induced to differentiate into THP-1 macrophages by phorbol 12-Myristate 13-Acetate(PMA,100.00μg/L).THP-1 monocyte-derived macrophages were divided into the control group(equal volume of culture medium)and different doses of ox-LDL groups(with 0.08 mg/L,0.80 mg/L,8.00 mg/L,and 80.00 mg/L).The toxicity of ox-LDL to THP-1-derived macrophages was detected by methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay.THP-1 macrophages were divided into the control group and 1,8-Cineole groups with different doses(1.00μmol/L,5.00μmol/L,25.00μmol/L,125.00μmol/L,and 625.00μmol/L),and and the effect of 1,8-Cineole on the activity of THP-1 derived macrophages was detected using MTT assay.THP-1 macrophages were induced to differentiate into foam cell models by ox-LDL with 80.00 mg/L,which were divided into the control group,ox-LDL group(80.00 mg/L ox-LDL),and ox-LDL group(80.00 mg/L ox-LDL)and 1,8-Cineole group[ox-LDL 80.00 mg/L+at various concentrations 1,8-Cineole(1.00μmol/L,5.00μmol/L,25.00μmol/L,125.00μmol/L,and 625.00μmol/L)].The effect of 1,8-Cineole on the activity of THP-1 foam cells induced by ox LDL was detected by MTT assay.THP-1 macrophages were induced by ox-LDL with 80.00 mg/L to differentiate THP-1 derived macrophages into foam cell models,and cells were divided into the control group(the same volume of medium),ox-LDL group(80.00 mg/L ox-LDL),low-dose 1,8-Cineole group(80.00 mg/L ox-LDL and 1.00μmol/L 1,8-Cineole),high-dose 1,8-Cineole group(80.00 mg/L ox-LDL and 5.00μmol/L 1,8-Cineole);Oil red O staining was used to detect the accumulation of lipids and the formation of foam cells;Nile red staining experiment was used to detect the cellular lipid uptake,Western Blot was used to analyze the expression of A1 scavenger receptor(SR-A1)protein in macrophages,and real-t

关 键 词：泡沫细胞 巨噬细胞 清道夫受体 A类 1 8-桉叶油素 氧化低密度脂蛋白 

分 类 号：R285.5[医药卫生—中药学]