灯盏乙素对OX-LDL损伤的RAW264.7细胞中PKC和TNF-α表达的影响  

Effects of Scutellarin on Expressions of PKC and TNF-αin OX-LDL Injured RAW264.7 Cells in vitro

作　　者：张莉 余丹 王雨婷 苟志梅 杨仁华 何波 沈志强 柴文英[2] 陈鹏 ZHANG Li;YU Dan;WANG Yu-ting;GOU Zhi-mei;YANG Ren-hua;HE Bo;SHEN Zhi-qiang;CHAI Wen-ying;CHEN Peng(School of Pharmaceutical Sciences and Yunnan Key Laboratory of Pharmacology for Natural Products,Kunming Medical University,Kunming Yunnan 650500;Dept,of Breast Surgery,First Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650031,China)

机构地区：[1]昆明医科大学药学院暨云南省天然药物药理重点实验室,云南昆明650500 [2]昆明医科大学第一附属医院乳腺外科,云南昆明6500031

出　　处：《昆明医科大学学报》2021年第10期1-7,共7页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(81860641);云南省基础研究计划重点项目(202001AS070035);昆明医科大学博士研究生创新基金资助项目(2021D05)。

摘　　要：目的研究灯盏乙素(scutellarin)对OX-LDL诱导损伤的巨噬细胞RAW264.7脂质吞噬能力及对PKC和TNF-α的蛋白和mRNA表达调控的影响。方法体外培养小鼠单核巨噬细胞RAW264.7,复制OXLDL诱导损伤模型,通过油红O染色检测脂质吞噬及试剂盒检测细胞损伤模型胆固醇酯的含量。Western blot和RT-PCR方法检测RAW264.7细胞中PKC和TNF-α表达变化。结果(1)油红O结果显示:与正常对照组比较,模型组细胞红染程度明显高于正常对照组;与模型组比较,辛伐他汀组、灯盏乙素50和100μmol/L组细胞红染程度均低于模型组;(2)胆固醇酯含量检测显示:与正常对照组比较,各时间点模型组细胞CE/TC值均明显升高;与模型组比较,各时间点用药组细胞CE/TC值均降低。(3)WB和RT-PCR结果显示:与正常对照组比较,模型组PKC和TNF-α的蛋白和mRNA表达升高;与模型组比较,辛伐他汀组(simvastatin)PKC和TNF-α的蛋白和mRNA表达降低,且灯盏乙素组呈浓度依赖性下调PKC和TNF-α的蛋白和mRNA表达。结论(1)灯盏乙素能明显抑制OX-LDL诱导损伤的RAW264.7细胞脂质吞噬能力。(2)灯盏乙素能够明显下调OX-LDL诱导损伤的RAW264.7中PKC和TNF-α的蛋白和mRNA表达水平。Objectives To investigate the effects of scutellarin on lipid phagocytosis and the regulation of PKC,and TNF-αexpressions in low density lipoprotein(OX-LDL)injured RAW264.7 cells.Methods Murine RAW264.7 macrophages was cultured in vitro.Then,the OX-LDL-induced injury model was replicated.Combined with oil red O staining,cellular cholesteryl ester content was also detected in OX-LDL-induced injury model.Furthermore,Western blot and RT-PCR methods were used to detect changes in PKC and TNF-αprotein as well as mRNA expression in RAW264.7 cells.Results(1)The results of oil red O showed that the degree of cell red staining in model group was significantly higher than that in normal control group;compared with model group,the degree of cell red staining in simvastatin group,scutellarin 50 and 100μmol/L group were lower than that in the model group;(2)The cholesteryl ester content assay showed that:compared with the normal control group,the CE/TC values of the model group cells were significantly increased at each time point;compared with model group,and cellular CE/TC values were significantly decreased in the dosing group at each time point.(3)WB and RT-PCR results showed that compared with normal control group,protein and m RNA expressions of PKC and TNF-αwere increased in the model group;compared with the model group,protein and m RNA expression of PKC and TNF-αwere decreased in simvastatin group,and scutellarin group showed a concentration-dependent downregulation of PKC and TNF-αexpression.Conclusions(1)Scutellarin can significantly inhibit the levels of cholesterol in OXLDL injuried RAW264.7 cells.(2)Scutellarin can down-regulated the protein and m RNA expressions of PKC,TNF-αin OX-LDL induced RAW264.7 cells.

关 键 词：灯盏乙素 巨噬细胞RAW264.7 氧化型低密度脂蛋白 蛋白激酶C 肿瘤坏死因子-α 

分 类 号：R966[医药卫生—药理学]