阿托伐他汀对HepG_2细胞中脂代谢相关基因表达的影响
|
摘要
目的:探讨阿托伐他汀对HepG_2细胞中载脂蛋白A1 (apoA1)天然反义转录(apoA1-NAT)的作用及对脂代谢相关基因表达的影响。方法:采用不同浓度阿托伐他汀(0、1、10和100nmol·L~(-1))干预HepG_2细胞,0nmol·L~(-1)阿托伐他汀组为对照组,在不同时间点(6、12、24和48h)提取各组HepG_2细胞总RNA,采用Real-time PCR法检测HepG_2细胞中apoA1-NAT和脂代谢相关基因mRNA表达水平。结果:1和10nmol·L~(-1)阿托伐他汀组HepG_2细胞形态正常。与6h时比较,12、24和48h时10nmol·L~(-1)阿托伐他汀组HepG_2细胞中apoA1-NAT表达水平明显降低(P<0.01),且呈明显的时间依赖性。在相同条件下,48h时HepG_2细胞中apoA1mRNA表达水平较6h时明显升高(P<0.01)。与对照组比较,100nmol·L~(-1)阿托代他汀组HepG_2细胞中低密度脂蛋白受体(LDLR)、10nmol·L~(-1)阿托代他汀组HepG_2细胞中清道夫受体B1(SRB1)及1和10nmol·L~(-1)阿托伐他汀组HepG_2细胞中ATP结合盒式转运体A1 (ABCA1)表达水平均有不同程度升高(P<0.01)。结论:阿托伐他汀通过抑制HepG_2细胞中apoA1-NAT表达,促进脂代谢相关基因表达,进而促进胆固醇逆转运过程。
Objective:To explore the effects of atorvastatin on the natural antisense transcription of apolipoprotein A1(apoA1-NAT)in the HepG_2 cells and its influence in the expressions of lipid metabolism related genes.Methods:The HepG_2 cells were intervened with different concentrations(0,1,10 and 100 nmol·L~(-1))of atorvastatin,and the 0 nmol·L~(-1) atorvastatin group was used as control group.The total RNA of HepG_2 cells in various groups were extracted at different time points(6,12,24,and 48 h).The mRNA expression levels of lipid metabolism-related genes and apoA1-NAT expression levels were detected by Real-Time PCR method.Results:The morphology of HepG_2 cells in 1 and 10 nmol·L~(-1) atorvastain groups was normal.Compared with 6 h,the expression levels of apoA1-NAT in the HepG_2 cells in 10 nmol·L~(-1) atorvastatin group at 12,24 and 48 hwere significantly decreased(P<0.01)in a time-dependent manner.Under the same condition,the expression level of apoA1 mRNA in the HepG_2 cells at 48 hwas increased significantly compared with 6 h(P<0.01).Compared with control group,the expression levels of low density lipoprotein receptor(LDLR)in the HepG_2 cells in100 nmol·L~(-1) atorvastatin group,scavenger receptor-class B type 1(SRB1)in 10 nmol·L~(-1) atorvastatin group and ATP binding cassette transporter A1(ABCA1)in 1 and 10 nmol·L~(-1) atorvastatin groups were increased(P<0.01)in different degrees.Conclusion:Atorvastatin can promote the expressions of lipid metabolism-related genes by inhibiting the expression of apoA1-NAT,and promote the process of reverse cholesterol transport.
Objective:To explore the effects of atorvastatin on the natural antisense transcription of apolipoprotein A1(apoA1-NAT)in the HepG_2 cells and its influence in the expressions of lipid metabolism related genes.Methods:The HepG_2 cells were intervened with different concentrations(0,1,10 and 100 nmol·L~(-1))of atorvastatin,and the 0 nmol·L~(-1) atorvastatin group was used as control group.The total RNA of HepG_2 cells in various groups were extracted at different time points(6,12,24,and 48 h).The mRNA expression levels of lipid metabolism-related genes and apoA1-NAT expression levels were detected by Real-Time PCR method.Results:The morphology of HepG_2 cells in 1 and 10 nmol·L~(-1) atorvastain groups was normal.Compared with 6 h,the expression levels of apoA1-NAT in the HepG_2 cells in 10 nmol·L~(-1) atorvastatin group at 12,24 and 48 hwere significantly decreased(P<0.01)in a time-dependent manner.Under the same condition,the expression level of apoA1 mRNA in the HepG_2 cells at 48 hwas increased significantly compared with 6 h(P<0.01).Compared with control group,the expression levels of low density lipoprotein receptor(LDLR)in the HepG_2 cells in100 nmol·L~(-1) atorvastatin group,scavenger receptor-class B type 1(SRB1)in 10 nmol·L~(-1) atorvastatin group and ATP binding cassette transporter A1(ABCA1)in 1 and 10 nmol·L~(-1) atorvastatin groups were increased(P<0.01)in different degrees.Conclusion:Atorvastatin can promote the expressions of lipid metabolism-related genes by inhibiting the expression of apoA1-NAT,and promote the process of reverse cholesterol transport.
引文
[1]KAPURIA D,TAKYAR V K,ETZION O,et al.Association of hepatic steatosis with subclinical atherosclerosis:systematic review and Meta-analysis[J].Hepatol Commun,2018,2(8):873-883.
[2]GAO S,ZHAO D,WANG M,et al.Association between circulating oxidized LDL and atherosclerotic cardiovascular disease:A Meta-analysis of observational studies[J].Can JCardiol,2017,33(12):1624-1632.
[3]FADAEI R,POUSTCHI H,MESHKANI R,et al.Impaired HDL cholesterol efflux capacity in patients with nonalcoholic fatty liver disease is associated with subclinical atherosclerosis[J].Sci Rep,2018,8(1):11691.
[4]KYPREOS K E,GKIZAS S,RALLIDIS L S,et al.HDLparticle functionality as a primary pharmacological target for HDL-based therapies[J].Biochem Pharmacol,2013,85(11):1575-1578.
[5]MORA S,GLYNN R J,RIDKER P M.High-density lipoprotein cholesterol,size,particle number,and residual vascular risk after potent statin therapy[J].Circulation,2013,128(11):1189-1197.
[6]WANG Y,NIIMI M,NISHIJIMA K,et al.Human Apolipoprotein A-Ⅱprotects against diet-Induced atherosclerosis in transgenic rabbits[J].Arterioscler Thromb Vasc Biol,2013,33(2):224-231.
[7]ROSENSON R S,BREWER H B,DAVIDSON W S,et al.Cholesterol efflux and atheroprotection:advancing the concept of reverse cholesterol transport[J].Circulation,2012,125(15):1905-1919.
[8]COOKE A L,MORRIS J,MELCHIOR J T,et al.Athumbwheel mechanism for APOA1 activation of LCATactivity in HDL[J].J Lipid Res,2018,59(7):1244-1255.
[9]UCHIDA S,DIMMELER S.Long noncoding RNAs in cardiovascular diseases[J].Circ Res,2015,116(4):737-750.
[10]HALLEY P,KADAKKUZHA B M,FAGHIHI M A,et al.Regulation of the apolipoprotein gene cluster by a long noncoding RNA[J].Cell Rep,2014,6(1):222-230.
[11]COVEY S D,KRIEGER M,WANG W,et al.Scavenger receptor class B typeⅠ-mediated protection against atherosclerosis in LDL receptor-negative mice involves its expression in bone marrow-derived cells[J].Arterioscler Thromb Vasc Biol,2003,23(9):1589-1594.
[12]RUBIN E M,KRAUSS R M,SPANGLER E A,et al.Inhibition of early atherogenesis in transgenic mice by human apolipoprotein AI[J].Nature,1991,353(6341):265-267.
[13]ZHANG Y,ZANOTTI I,REILLY M P,et al.Overexpression of apolipoprotein A-I promotes reverse transport of cholesterol from macrophages to feces in vivo[J].Circulation,2003,108(6):661-663.
[14]DUVERGER N,KRUTH H,EMMANUEL F,et al.Inhibition of atherosclerosis development in cholesterol-fed human apolipoprotein A-I-transgenic rabbits[J].Circulation,1996,94(4):713-717.
[15]IKENAGA M,HIGAKI Y,SAKU K,et al.High-density lipoprotein mimetics:a therapeutic tool for atherosclerotic diseases[J].J Atheroscler Thromb,2016,23(4):385-394.
[16]FAGHIHI M A,WAHLESTEDT C.Regulatory roles of natural antisense transcripts[J].Nat Rev Mol Cell Biol,2009,10(9):637-643.
[17]WAHLESTEDT C.Natural antisense and noncoding RNAtranscripts as potential drug targets[J].Drug Discov Today,2006,11(11/12):503-508.
[18]LEHNER B,WILLIAMS G,CAMPBELL R D,et al.Antisense transcripts in the human genome[J].Trends Genet,2002,18(2):63-65.
[19]张鹏,王艳丽,郭世文,等.家兔胆固醇酯转移蛋白基因克隆及其真核表达载体的构建[J].吉林大学学报:医学版,2013,39(4):837-840.
[20]SHRESTHA S,WU B J,GUINEY L,et al.Cholesteryl ester transfer protein and its inhibitors[J].J Lipid Res,2018,59(5):772-783.
[2]GAO S,ZHAO D,WANG M,et al.Association between circulating oxidized LDL and atherosclerotic cardiovascular disease:A Meta-analysis of observational studies[J].Can JCardiol,2017,33(12):1624-1632.
[3]FADAEI R,POUSTCHI H,MESHKANI R,et al.Impaired HDL cholesterol efflux capacity in patients with nonalcoholic fatty liver disease is associated with subclinical atherosclerosis[J].Sci Rep,2018,8(1):11691.
[4]KYPREOS K E,GKIZAS S,RALLIDIS L S,et al.HDLparticle functionality as a primary pharmacological target for HDL-based therapies[J].Biochem Pharmacol,2013,85(11):1575-1578.
[5]MORA S,GLYNN R J,RIDKER P M.High-density lipoprotein cholesterol,size,particle number,and residual vascular risk after potent statin therapy[J].Circulation,2013,128(11):1189-1197.
[6]WANG Y,NIIMI M,NISHIJIMA K,et al.Human Apolipoprotein A-Ⅱprotects against diet-Induced atherosclerosis in transgenic rabbits[J].Arterioscler Thromb Vasc Biol,2013,33(2):224-231.
[7]ROSENSON R S,BREWER H B,DAVIDSON W S,et al.Cholesterol efflux and atheroprotection:advancing the concept of reverse cholesterol transport[J].Circulation,2012,125(15):1905-1919.
[8]COOKE A L,MORRIS J,MELCHIOR J T,et al.Athumbwheel mechanism for APOA1 activation of LCATactivity in HDL[J].J Lipid Res,2018,59(7):1244-1255.
[9]UCHIDA S,DIMMELER S.Long noncoding RNAs in cardiovascular diseases[J].Circ Res,2015,116(4):737-750.
[10]HALLEY P,KADAKKUZHA B M,FAGHIHI M A,et al.Regulation of the apolipoprotein gene cluster by a long noncoding RNA[J].Cell Rep,2014,6(1):222-230.
[11]COVEY S D,KRIEGER M,WANG W,et al.Scavenger receptor class B typeⅠ-mediated protection against atherosclerosis in LDL receptor-negative mice involves its expression in bone marrow-derived cells[J].Arterioscler Thromb Vasc Biol,2003,23(9):1589-1594.
[12]RUBIN E M,KRAUSS R M,SPANGLER E A,et al.Inhibition of early atherogenesis in transgenic mice by human apolipoprotein AI[J].Nature,1991,353(6341):265-267.
[13]ZHANG Y,ZANOTTI I,REILLY M P,et al.Overexpression of apolipoprotein A-I promotes reverse transport of cholesterol from macrophages to feces in vivo[J].Circulation,2003,108(6):661-663.
[14]DUVERGER N,KRUTH H,EMMANUEL F,et al.Inhibition of atherosclerosis development in cholesterol-fed human apolipoprotein A-I-transgenic rabbits[J].Circulation,1996,94(4):713-717.
[15]IKENAGA M,HIGAKI Y,SAKU K,et al.High-density lipoprotein mimetics:a therapeutic tool for atherosclerotic diseases[J].J Atheroscler Thromb,2016,23(4):385-394.
[16]FAGHIHI M A,WAHLESTEDT C.Regulatory roles of natural antisense transcripts[J].Nat Rev Mol Cell Biol,2009,10(9):637-643.
[17]WAHLESTEDT C.Natural antisense and noncoding RNAtranscripts as potential drug targets[J].Drug Discov Today,2006,11(11/12):503-508.
[18]LEHNER B,WILLIAMS G,CAMPBELL R D,et al.Antisense transcripts in the human genome[J].Trends Genet,2002,18(2):63-65.
[19]张鹏,王艳丽,郭世文,等.家兔胆固醇酯转移蛋白基因克隆及其真核表达载体的构建[J].吉林大学学报:医学版,2013,39(4):837-840.
[20]SHRESTHA S,WU B J,GUINEY L,et al.Cholesteryl ester transfer protein and its inhibitors[J].J Lipid Res,2018,59(5):772-783.