摘要
目的:从巨噬细胞泡沫化角度探讨温胆汤抗动脉粥样硬化的作用机制。方法:采用MTT法确定温胆汤的无毒性干预浓度;药物干预后,采用油红O染色法检测泡沫细胞的形成;采用酶法检测细胞内胆固醇水平及细胞胆固醇流出;采用Western blot检测巨噬细胞膜蛋白CD36、A类清道夫受体(SR-A)及ATP结合盒转运体A1(ABCA1)和B类I型清道夫受体(SR-BI)的蛋白表达水平。结果:温胆汤冻干粉溶液对氧化型低密度脂蛋白(ox-LDL)刺激的THP-1细胞源性巨噬细胞的无毒性干预浓度为0~6 g/L,于是设置温胆汤干预浓度梯度为1.5、3和6 g/L进行后续实验。各浓度温胆汤干预可抑制ox-LDL诱导的泡沫细胞形成,半定量分析显示温胆汤浓度依赖性降低细胞内脂质沉积(P<0.05或P<0.01)。温胆汤浓度依赖性降低细胞内总胆固醇、胆固醇酯和胆固醇酯化率(P<0.05或P<0.01),促进细胞胆固醇流出(P<0.01),减少CD36表达(P<0.01)。6 g/L温胆汤显著减少SR-A的表达(P<0.05);3和6 g/L温胆汤显著促进ABCA1和SR-BI的表达(P<0.05或P<0.01)。结论:温胆汤显著抑制巨噬细胞泡沫化,相关机制可能是通过抑制巨噬细胞膜蛋白CD36和SR-A表达而减少巨噬细胞对胆固醇的内化吞噬,同时上调巨噬细胞膜蛋白ABCA1和SR-BI表达,从而促进巨噬细胞内胆固醇流出。
AIM:To explore the anti-atherosclerotic mechanism of Wendan decoction based on formation of foam cells.METHODS:The optimal concentrations of Wendan decoction without cytotoxity to cells were selected by MTT assay.After Wendan decoction treatment,the formation of foam cells was examined by oil red O staining.The cho⁃lesterol efflux,cholesterol level,free cholesterol level and cholesterol esterification rate were analyzed using cholesterol ef⁃flux assay,total cholesterol assay and free cholesterol assay.The expression levels of macrophage membrane proteins,in⁃cluding CD36,scavenger receptor class A(SR-A),ATP-binding cassette transporter A1(ABCA1)and scavenger recep⁃tor class B type I(SR-BI),were quantified by Western blot.RESULTS:The optimal concentrations of Wendan decoc⁃tion without cytotoxity to the cells were 0~6 g/L.Wendan decoction at the concentrations of 1.5,3 and 6 g/L were selected for the experiments.Wendan decoction at these concentrations inhibited the formation of foam cells induced by oxidized low-density lipoprotein(ox-LDL),and reduced the accumulation of intracellular lipids in a concentration-dependent man⁃ner(P<0.05 or P<0.01).Wendan decoction also reduced intracellular total cholesterol level,cholesterol ester level and cholesterol esterification rate(P<0.05 or P<0.01),promoted efflux of intracellular cholesterol(P<0.01),and decreased the protein level of CD36 in THP-1 cell-derived macrophages(P<0.01)in a concentration-dependent manner.Wendan decoction at the concentration of 6 g/L significantly reduced the protein level of SR-A in THP-1 cell-derived macrophages(P<0.05).At the concentrations of 3 and 6 g/L,Wendan decoction significantly increased the protein levels of ABCA1 and SR-BI in THP-1 cell-derived macrophages(P<0.05 or P<0.01).CONCLUSION:Wendan decoction significantly inhibits ox-LDL-induced formation of foam cells by reducing cholesterol deposition and promoting cholesterol efflux,and its mechanism may be related to the down-regulation of CD36 and SR-A and the up-regulation
作者
陈玄晶
钟碧莹
吴焕林
徐丹苹
CHEN Xuan-jing;ZHONG Bi-ying;WU Huan-lin;XU Dan-ping(The Second Clinical Medical College of Guangzhou University of Chinese Medicine;Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510020,China;Beijing University of Chinese Medicine,Beijing 100700,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第11期1952-1959,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81774219)
中医药科学技术研究专项(No.YN2019MJ13)。
