石榴皮多酚对胆固醇流出及相关蛋白表达的影响
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摘要
利用Raw264.7巨噬细胞源性泡沫细胞模型,阐明石榴皮多酚对高密度脂蛋白(High density lipoprotein,HDL)介导Raw264.7泡沫细胞胆固醇流出的调控作用,并采用蛋白质免疫印迹法(Western blotting,WB),分析石榴皮多酚对巨噬细胞胆固醇流出相关蛋白清道夫受体B1(Scavenger receptor B1,SR-B1)、酰基辅酶A1:胆固醇酰基转移酶(acyl-coenzyme A1:cholesterol acyltransferase,ACAT1)和胆固醇酯水解酶(cholesteryl ester hydrolase,nCEH)表达的影响,揭示细胞ACAT1和nCEH活性的变化。结果表明:石榴皮多酚可促进HDL介导的胆固醇流出,显著上调胆固醇流出相关蛋白SRB1的表达(P<0.05);此外,石榴皮多酚对nCEH和ACAT1蛋白表达影响不显著(P>0.05),但可显著降低ACAT1的酶活(P<0.05),提高nCEH酶活性(P<0.05),从而有利于细胞内胆固醇酯向游离胆固醇的转化和流出。
In order to explore the effects of pomegranate peel polyphenols(PPPs)on SR-B1 protein expression and cholesterol efflux,in this study,Raw264.7 macrophages derived from foam cells were used as the model to illuminate the effects of PPPs on cholesterol efflux mediated by HDL.Western blot method was used to analyze the expression of scavenger receptor B1(SR-B1),acyl-coenzyme A1:cholesterol acyltransferase(ACAT1)and cholesteryl ester hydrolase(nCEH).Further,the activity changes of ACAT1 and nCEH were revealed.The results showed that PPPs promoted the HDL-mediated cholesterol efflux and upregulated the expression of SR-B1 significantly(P<0.05).In addition,the PPPs had no significant effects on nCEH and ACAT1 protein expression(P>0.05),but could significantly reduce the enzyme activity of ACAT1 and increase that of nCEH(P<0.05),which facilitated the transformation and outflow of intracellular cholesterol esters to free cholesterol.
In order to explore the effects of pomegranate peel polyphenols(PPPs)on SR-B1 protein expression and cholesterol efflux,in this study,Raw264.7 macrophages derived from foam cells were used as the model to illuminate the effects of PPPs on cholesterol efflux mediated by HDL.Western blot method was used to analyze the expression of scavenger receptor B1(SR-B1),acyl-coenzyme A1:cholesterol acyltransferase(ACAT1)and cholesteryl ester hydrolase(nCEH).Further,the activity changes of ACAT1 and nCEH were revealed.The results showed that PPPs promoted the HDL-mediated cholesterol efflux and upregulated the expression of SR-B1 significantly(P<0.05).In addition,the PPPs had no significant effects on nCEH and ACAT1 protein expression(P>0.05),but could significantly reduce the enzyme activity of ACAT1 and increase that of nCEH(P<0.05),which facilitated the transformation and outflow of intracellular cholesterol esters to free cholesterol.
引文
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[13]ZHAO Sheng-juan,LI Jian-ke,WANG Li-fang,et al.Pomegranate peel polyphenols inhibit lipid accumulation and enhance cholesterol efflux in raw264.7 macrophages[J].Food&Function,2016,7(7):3 201-3 210.
[14]周云,沃兴德,卢德赵.RAW264.7巨噬细胞源性泡沫细胞模型的建立及鉴定[J].中国动脉硬化杂志,2010,18(9):687-690.
[15]刘庆艳,白静,赵宇,等.白细胞介素34对巨噬细胞泡沫化过程中细胞内胆固醇水平的影响[J].中华老年心脑血管病杂志,2017,19(1):16-19.
[16]何雪峰,李晓辉,李淑慧,等.三七总皂苷对小鼠巨噬细胞源性泡沫细胞形成的影响[J].中国药房,2007,18(30):2 323-2 325.
[2]MOORE K J,SHEEDY F J,FISHER E A.Macrophages in atherosclerosis:a dynamic balance[J].Nature Reviews Immunology,2013,13:709-721.
[3]GLASS C K,WITZTUM J L.Atherosclerosis:the road ahead[J].Cell,2001,104(4):503-516.
[4]徐留建,杨小东,张剑梅,等.细胞胆固醇流出通路研究进展[J].中国心血管病研究,2012,10(4):309-312.
[5]梁俊.石榴皮多酚对脂质过氧化及肝细胞HMG-CoA还原酶活性及表达的影响[D].西安:陕西师范大学,2012:1-3.
[6]ROM O,AVIRAM M.Paraoxsonase2(PON2)and oxidative stress involvement in pomegranate juice protection against cigarette smoke-induced macrophage cholesterol accumulation[J].Chemico-Biological Interactions,2016,259(Pt B):394-400.
[7]马倩倩.石榴皮多酚对肝细胞胆固醇合成影响及其分子机制研究[D].西安:陕西师范大学,2013:21-26.
[8]QIN Li,YANG Yun-bo,YANG Yi-xin,et al.Inhibition of macrophage-derived foam cell formation by ezetimibe via the caveolin-1/MAPK pathway[J].Clinical and Experimental Pharmacology and Physiology,2016,43(2):182-192.
[9]JAMES R W,BRULHART-MEYNET M C,SINGH A K,et al.The scavenger receptor class B,type I is a primary determinant of paraoxonase-1association with high-density lipoproteins[J].Arteriosclerosis Thrombosis&Vascular Biology,2010,30(11):2 121-2 127.
[10]弓伊宁,李芙蓉,姚伟娟.巨噬细胞的胆固醇代谢及其影响因素[J].生理科学进展,2016,47(4):291-294.
[11]SEKIYA M,OSUGA J,IGARASHI M,et al,Ishibashi SThe role of neutral cholesterol ester hydrolysis in macrophage foam cells[J].Journal of Atherosclerosis and Thrombosis,2011,18(5):359-364.
[12]DURHAM K K,CHATHELY K M,MAK K C,et al.HDL protects against doxorubicin-induced cardiotoxicity in a scavenger receptor class B type 1-,PI3K-,and Akt-dependent manner[J].American Journal of Physiology-Heart and Circulatory Physiology,2018,314(1):H31-H44.
[13]ZHAO Sheng-juan,LI Jian-ke,WANG Li-fang,et al.Pomegranate peel polyphenols inhibit lipid accumulation and enhance cholesterol efflux in raw264.7 macrophages[J].Food&Function,2016,7(7):3 201-3 210.
[14]周云,沃兴德,卢德赵.RAW264.7巨噬细胞源性泡沫细胞模型的建立及鉴定[J].中国动脉硬化杂志,2010,18(9):687-690.
[15]刘庆艳,白静,赵宇,等.白细胞介素34对巨噬细胞泡沫化过程中细胞内胆固醇水平的影响[J].中华老年心脑血管病杂志,2017,19(1):16-19.
[16]何雪峰,李晓辉,李淑慧,等.三七总皂苷对小鼠巨噬细胞源性泡沫细胞形成的影响[J].中国药房,2007,18(30):2 323-2 325.
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