绞股蓝总皂甙对肝HepG2细胞三酰甘油代谢的影响及机制
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摘要
目的探讨绞股蓝总皂甙(GPs)对培养肝HepG2细胞三酰甘油(TG)代谢的影响及机制。方法采用HepG2细胞为模型细胞,加入GPs进行孵育,观察其对细胞内TG的影响,并采用荧光定量PCR的方法,分析脂代谢过程中的关键酶基因,包括乙酰辅酶A羧化酶(ACAC)A、ACACB、脂肪酸合成酶(FASN)、乙酰辅酶A乙酰转移酶(ACAT1)、棕榈酰辅酶A氧化酶(ACOX1)及肉碱脂酰转移酶(CPT)1、2。并进一步在细胞水平干扰关键酶基因,验证其对TG代谢的影响。结果 GPs可显著降低HepG2细胞的TG水平,发现GPs可显著抑制FASN基因的表达及其在较高水平下促进CPT1基因的表达,其蛋白水平的表达改变与基因表达一致。在细胞水平瞬时干扰FASN的表达,在干扰FASN后,GPs降低TG的幅度显著减少,提示GPs主要通过抑制FASN起作用。结论 GPs可减少肝HepG2细胞内TG水平,其作用主要是通过抑制FASN的表达而抑制脂肪酸的合成。
Objective To explore the effect and mechanism of gypenosides(GPs)on triglycerides(TG)metabolism in cultured liver HepG2 cells.Methods The HepG2 cells were adopted as the model cells and incubated with GPs.Their effect on intracellular triglycerides was observed.The fluorescent quantitative RT-PCR was adopted to analyze the expression of key enzyme genes in lipid metabolism including acetyl-CoA carboxylase(ACACA,ACACB),fatty acid synthase(FASN),acetyl-CoA acetyltransferase 1(ACAT1),acyl-CoA palmitoyl oxidase 1(ACOX1),carnitine palmitoyltransferase 1(CPT1)and carnitine palmitoyltransferase 2(CPT2).Then the effect on TG metabolism was further verified by interfering the key enzyme gene at the cellular level.Results GPs could significantly decrease the level of intracellular triglycerides in HepG2 cells,and could significantly inhibit the expression of FASN gene and promote the expression of CPT1 gene in a higher concentration,and its protein level expression was consistent with the gene expression.Moreover,the expression of FASN was transiently interfered at the cellular level,after interfering FASN,the magnitude of GPs in decreasing TG was significantly decreased,suggesting that GPs played the effect mainly by inhibiting FASN.Conclusion GPs could decrease the intracellular TG level of liver HepG2,its effect in inhibition of the synthesis of fatty acid is mainly through the inhibition of FASN.
Objective To explore the effect and mechanism of gypenosides(GPs)on triglycerides(TG)metabolism in cultured liver HepG2 cells.Methods The HepG2 cells were adopted as the model cells and incubated with GPs.Their effect on intracellular triglycerides was observed.The fluorescent quantitative RT-PCR was adopted to analyze the expression of key enzyme genes in lipid metabolism including acetyl-CoA carboxylase(ACACA,ACACB),fatty acid synthase(FASN),acetyl-CoA acetyltransferase 1(ACAT1),acyl-CoA palmitoyl oxidase 1(ACOX1),carnitine palmitoyltransferase 1(CPT1)and carnitine palmitoyltransferase 2(CPT2).Then the effect on TG metabolism was further verified by interfering the key enzyme gene at the cellular level.Results GPs could significantly decrease the level of intracellular triglycerides in HepG2 cells,and could significantly inhibit the expression of FASN gene and promote the expression of CPT1 gene in a higher concentration,and its protein level expression was consistent with the gene expression.Moreover,the expression of FASN was transiently interfered at the cellular level,after interfering FASN,the magnitude of GPs in decreasing TG was significantly decreased,suggesting that GPs played the effect mainly by inhibiting FASN.Conclusion GPs could decrease the intracellular TG level of liver HepG2,its effect in inhibition of the synthesis of fatty acid is mainly through the inhibition of FASN.
引文
[1]金亭亭,孙兆林,江蔚新.绞股蓝化学成分及药理作用研究进展[J].亚太传统医药,2014,10(16):30-32.
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[9]Yan H,Wang X,Wang Y,et al.Antiproliferation and anti-migration induced by gypenosides in human colon cancer SW620and esophageal cancer Eca-109cells[J].Hum Exp Toxicol,2014,33(5):522-533.
[10]Lu HF,Chen YS,Yang JS,et al.Gypenosides induced G0/G1arrest via inhibition of cyclin E and induction of apoptosis via activation of caspases-3and-9in human lung cancer A-549cells[J].In Vivo,2008,22(2):215-221.
[11]Liu JS,Chiang TH,Wang JS,et al.Induction of p53-independent growth inhibition in lung carcinoma cell A549by gypenosides[J].J Cell Mol Med,2015,19(7):1697-1709.
[12]Sun DP.Gypenosides induce apoptosis by Ca2+overload mediated by endoplasmic-reticulum and store-operated Ca2+channels in human hepatoma cells[J].Cancer Bio ther Radiopharm,2013,28(4):320-326.
[13]Shi L,Pi Y,Luo C,et al.In vitro inhibitory activities of six gypenosides on human liver cancer cell line HepG2and possible role of HIF-1αpathway in them[J].Chem Biol Interact,2015,238(1):48-54.
[14]Tong L.Acetyl-coenzyme A carboxylase:crucial metabo lic enzyme and attractive target for drug discovery[J].Cell Mol Life Sci,2005,62(16):1784-1803.
[15]Peng Y,Lei T,Yuan J,et al.Arachidonic acid induces acetyl-CoA carboxylase 1 expression via activation of CREB1[J].Endocrine,2009,36(3):491-497.
[2]沈楠,许文频,李敏,等.绞股蓝皂苷对高脂血症大鼠脂代谢的影响[J].中西医结合心脑血管病杂志,2011,9(9):1081-1083.
[3]贺琴,谭华炳,赵琴.绞股蓝总皂甙抗高脂血症致动脉粥样硬化的机制研究进展[J].中西医结合心脑血管病杂志,2009,7(4):464-465.
[4]汪鋆植,胡格,翟文海.绞股蓝总皂苷外用对高血脂小鼠的降血脂作用研究[J].中国民族民间医药杂志,2007,16(1):41-43.
[5]王亚利,范春雷,窦晓兵.绞股蓝总皂苷对肝细胞低密度脂蛋白受体表达的影响[J].中国药理学通报,2010,26(1):138-139.
[6]Qin R,Zhang J,Li C,et al.Protective effects of gypenosides against fatty liver disease induced by high fat and cholesterol diet and alcohol in rats[J].Arch Pharm Res,2012,35(7):1241-1250.
[7]Chen JC,Lu KW,Tsai ML,et al.Gypenosides induced G0/G1arrest via CHk2and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4cells[J].Oral Oncol,2009,45(3):273-283.
[8]Lu KW,Tsai ML,Chen JC,et al.Gypenosides inhibited invasion and migration of human tongue cancer SCC4cells through down-regulation of NFkappaB and matrix metalloproteinase-9[J].Anticancer Res,2008,28(2A):1093-1099.
[9]Yan H,Wang X,Wang Y,et al.Antiproliferation and anti-migration induced by gypenosides in human colon cancer SW620and esophageal cancer Eca-109cells[J].Hum Exp Toxicol,2014,33(5):522-533.
[10]Lu HF,Chen YS,Yang JS,et al.Gypenosides induced G0/G1arrest via inhibition of cyclin E and induction of apoptosis via activation of caspases-3and-9in human lung cancer A-549cells[J].In Vivo,2008,22(2):215-221.
[11]Liu JS,Chiang TH,Wang JS,et al.Induction of p53-independent growth inhibition in lung carcinoma cell A549by gypenosides[J].J Cell Mol Med,2015,19(7):1697-1709.
[12]Sun DP.Gypenosides induce apoptosis by Ca2+overload mediated by endoplasmic-reticulum and store-operated Ca2+channels in human hepatoma cells[J].Cancer Bio ther Radiopharm,2013,28(4):320-326.
[13]Shi L,Pi Y,Luo C,et al.In vitro inhibitory activities of six gypenosides on human liver cancer cell line HepG2and possible role of HIF-1αpathway in them[J].Chem Biol Interact,2015,238(1):48-54.
[14]Tong L.Acetyl-coenzyme A carboxylase:crucial metabo lic enzyme and attractive target for drug discovery[J].Cell Mol Life Sci,2005,62(16):1784-1803.
[15]Peng Y,Lei T,Yuan J,et al.Arachidonic acid induces acetyl-CoA carboxylase 1 expression via activation of CREB1[J].Endocrine,2009,36(3):491-497.