HCV对脂代谢相关指标表达的影响
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摘要
目的研究HCV在体内外对脂代谢相关指标表达的影响。方法选取武汉大学人民医院2017年10月-2018年10月HCV感染者的标本114例(试验组)和健康体检者标本96例(对照组),采用全自动生化分析仪检测血脂指标,包括甘油三酯、总胆固醇、高密度脂蛋白、低密度脂蛋白、小而低密度脂蛋白、脂蛋白a、载脂蛋白A1和载脂蛋白B的血清水平;采用实时荧光定量PCR检测HCV感染的Huh7. 5. 1细胞(实验细胞)和未感染HCV的Huh7. 5. 1细胞(对照细胞)中脂肪酸合成酶、乙酰CoA羧化酶、羟甲基戊二酸单酰CoA还原酶、载脂蛋白A1、载脂蛋白B和低密度脂蛋白受体mRNA的表达量。正态分布的计量资料2组间比较采用t检验;非正态分布的计量资料2组间比较采用Mann-Whitney U检验。结果试验组患者血清总胆固醇、高密度脂蛋白、低密度脂蛋白、载脂蛋白A1和载脂蛋白B水平均低于对照组[分别为(2. 98±0. 51) mmol/L vs (4. 24±0. 43) mmol/L、(0. 87±0. 16) mmol/L vs (1. 24±0. 21) mmol/L、(1. 75±0. 24) mmol/L vs (2. 64±0. 37) mmol/L、(0. 94±0. 18) mmol/L vs (1. 47±0. 26) mmol/L、(0. 67±0. 31) mmol/L vs (0. 98±0. 14) mmol/L],差异均有统计学意义(t值分别为4. 96、5. 65、3. 88、3. 71、4. 41,P值均<0. 05)。实验细胞中载脂蛋白A1 mRNA表达水平较对照细胞低,差异有统计学意义(t=-3. 43,P <0. 05);脂肪酸合成酶、乙酰CoA羧化酶、羟甲基戊二酸单酰CoA还原酶、载脂蛋白B和低密度脂蛋白受体的mRNA表达水平均较对照细胞高,差异均有统计学意义(t值分别为5. 40、4. 93、3. 34、6. 88、3. 84,P值均<0. 05)。结论 HCV感染能够上调肝细胞内脂肪酸和胆固醇合成相关酶类基因mRNA水平,在体内和体外影响机体脂代谢。
Objective To investigate the influence of hepatitis C virus( HCV) on the expression of lipid metabolism indices in vitro and in vivo. Methods A total of 114 samples of patients with HCV infection who were treated in Renmin Hospital of Wuhan University from September 2017 to September 2018 were collected as experimental group,and 96 samples of healthy individuals who underwent physical examination were collected as control group. An automatic biochemical analyzer was used to measure blood lipid parameters,including triglyceride( TG),total cholesterol( TCh),high-density lipoprotein( HDL),low-density lipoprotein( LDL),small and low-density lipoprotein( sdLDL),lipoprotein( a),apolipoprotein A1( ApoA1),and apolipoprotein B( ApoB). RT-q PCR was used to measure the mRNA expression of fatty acid synthase( FASN),acetyl-CoA carboxylase( ACACA),hydroxymethylglutarate mono-acyl CoA reductase( HMGR),ApoA1,ApoB,and low-density lipoprotein receptor( LDLR) in Huh7. 5. 1 cells with or without HCV infection. The t-test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. Results Compared with the control group,the experimental group had significantly lower serum levels of TCh( 2. 98 ± 0. 51 mmol/L vs 4. 24 ± 0. 43 mmol/L,t = 4. 96,P < 0. 05),HDL( 0. 87 ± 0. 16 mmol/L vs 1. 24 ± 0. 21 mmol/L,t = 5. 65,P < 0. 05),LDL( 1. 75 ± 0. 24 mmol/L vs 2. 64 ± 0. 37 mmol/L,t = 3. 88,P < 0. 05),ApoA1( 0. 94± 0. 18 mmol/L vs 1. 47 ± 0. 26 mmol/L,t = 3. 71,P < 0. 05),and ApoB( 0. 67 ± 0. 31 mmol/L vs 0. 98 ± 0. 14 mmol/L,t = 4. 41,P <0. 05). Huh7. 5. 1 cells with HCV infection had significantly lower mRNA expression of Apo A1 than those without HCV infection( t =-3. 43,P < 0. 05),as well as significantly higher mRNA expression of FASN,ACACA,HMGR,ApoB,and LDLR( t = 5. 40,4. 93,3. 34,6. 88,and 3. 84,all P < 0. 05). Conclusion HCV infection can upregulate the mRNA levels of enzymes involved in the synthesis of fatty acids and cholesterol and thus affect lipid metabolism in vivo and in vitro.
Objective To investigate the influence of hepatitis C virus( HCV) on the expression of lipid metabolism indices in vitro and in vivo. Methods A total of 114 samples of patients with HCV infection who were treated in Renmin Hospital of Wuhan University from September 2017 to September 2018 were collected as experimental group,and 96 samples of healthy individuals who underwent physical examination were collected as control group. An automatic biochemical analyzer was used to measure blood lipid parameters,including triglyceride( TG),total cholesterol( TCh),high-density lipoprotein( HDL),low-density lipoprotein( LDL),small and low-density lipoprotein( sdLDL),lipoprotein( a),apolipoprotein A1( ApoA1),and apolipoprotein B( ApoB). RT-q PCR was used to measure the mRNA expression of fatty acid synthase( FASN),acetyl-CoA carboxylase( ACACA),hydroxymethylglutarate mono-acyl CoA reductase( HMGR),ApoA1,ApoB,and low-density lipoprotein receptor( LDLR) in Huh7. 5. 1 cells with or without HCV infection. The t-test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. Results Compared with the control group,the experimental group had significantly lower serum levels of TCh( 2. 98 ± 0. 51 mmol/L vs 4. 24 ± 0. 43 mmol/L,t = 4. 96,P < 0. 05),HDL( 0. 87 ± 0. 16 mmol/L vs 1. 24 ± 0. 21 mmol/L,t = 5. 65,P < 0. 05),LDL( 1. 75 ± 0. 24 mmol/L vs 2. 64 ± 0. 37 mmol/L,t = 3. 88,P < 0. 05),ApoA1( 0. 94± 0. 18 mmol/L vs 1. 47 ± 0. 26 mmol/L,t = 3. 71,P < 0. 05),and ApoB( 0. 67 ± 0. 31 mmol/L vs 0. 98 ± 0. 14 mmol/L,t = 4. 41,P <0. 05). Huh7. 5. 1 cells with HCV infection had significantly lower mRNA expression of Apo A1 than those without HCV infection( t =-3. 43,P < 0. 05),as well as significantly higher mRNA expression of FASN,ACACA,HMGR,ApoB,and LDLR( t = 5. 40,4. 93,3. 34,6. 88,and 3. 84,all P < 0. 05). Conclusion HCV infection can upregulate the mRNA levels of enzymes involved in the synthesis of fatty acids and cholesterol and thus affect lipid metabolism in vivo and in vitro.
引文
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[12] TIAN Y,YANG Y,ZHANG X,et al. Age-dependent PPARαactivation induces hepatic sulfatide accumulation in transgenic mice carrying the hepatitis C virus core gene[J]. Glycoconj J,2016,33(6):927-936.
[13] MURTHY GD,VU K,VENUGOPAL S. Prevalence and treatment of hyperlipidemia in patients with chronic hepatitis C infection[J]. Eur J Gastroenterol Hepatol,2009,21(8):902-907.
[14] HSU CS,LIU CH,LIU CJ,et al. Association of lipid profiles with hepatitis C viral load in chronic hepatitis C patients with genotype1 or 2 infection[J]. Am J Gastroenterol,2009,104(3):598.
[15] DAI CY,CHUANG WL,HO CK,et al. Associations between hepatitis C viremia and low serum triglyceride and cholesterol levels:A community-based study[J]. J Hepatology,2008,49(1):9-16.
[16] COREY KE,KANE E,MUNROE C,et al. Hepatitis C virus infection and its clearance alter circulating lipids:Implications for long-term follow-up[J]. Hepatology,2009,50(4):1030-1037.
[17] MIRANDOLA S,BOWMAN D,HUSSAIN MM,et al. Hepatic steatosis in hepatitis C is a storage disease due to HCV interaction with microsomal triglyceride transfer protein(MTP)[J].Nutr Metab(Lond),2010,7:13.
[18] TSAI TY,WANG WT,LI HK,et al. RNA helicase DDX3 maintains lipid homeostasis through upregulation of the microsomal triglyceride transfer protein by interacting with HNF4 and SHP[J]. Sci Rep,2017,7:41452.
[19] MOGILENKO DA,DIZHE EB,SHAVVA VS,et al. Role of the nuclear receptors HNF4α,PPARα,and LXRs in the TNFα-mediated inhibition of human apolipoprotein A-I gene expression in HepG2 cells[J]. Biochemistry,2009,48(50):11950-11960.
[2] SHIMIZU K,SOROIDA Y,SATO M,et al. Eradication of hepatitis C virus is associated with the attenuation of steatosis as evaluated using a controlled attenuation parameter[J]. Sci Rep,2018,8(1):7845.
[3] SORESI M,TRIPI S,FRANCO V,et al. Impact of liver steatosis on the antiviral response in the hepatitis C virus-associated chronic hepatitis[J]. Liver Int,2006,26(9):1119-1125.
[4] Chinese Society of Hepatology and Chinese Society of Infectious Diseases,Chinese Medical Association. The guideline of prevention and treatment for hepatitis C:A 2015 update[J]. J Clin Hepatol,2015,31(12):1961-1979.(in Chinese)中华医学会肝病学分会,中华医学会感染病学分会.丙型肝炎防治指南(2015年更新版)[J].临床肝胆病杂志,2015,31(12):1961-1979.
[5] PFAFFL MW. A new mathematical model for relative quantification in real-time RT-PCR[J]. Nucleic Acids Res,2001,29(9):e45.
[6] LIVAK KJ,SCHMITTGEN TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))method[J]. Methods,2001,25(4):402-408.
[7] ZHOU Y,YING L,XU JJ,et al. Effects of soluble human leukocyte antigen G on the standardized treatment of hepatitis C patients[J]. Chin J Clin Pharmacol Ther,2018,23(7):809-813.(in Chinese)周勇,应莉,徐佳佳,等.可溶性人类白细胞抗原G对丙型肝炎初治患者标准化治疗疗效的影响[J].中国临床药理学与治疗学,2018,23(7):809-813.
[8] CHENG DY,LIU XM,OU WN,et al. Clinical efficacy of directacting antiviral agents in patients with HCV-related liver cirrhosis[J/CD]. Chin J Exp Clin Infect Dis:Electronic Edition,2017,11(6):545-549.(in Chinese)程丹颖,刘晓民,欧蔚妮,等.直接抗病毒药物治疗丙型肝炎肝硬化患者的疗效观察[J/CD].中华实验和临床感染病杂志:电子版,2017,11(6):545-549.
[9] MARTIN NK,VICKERMAN P,DORE GJ,et al. Comparing individual and population prevention benefits of early directacting antiviral treatment for HCV[J]. J Hepatol,2016,64(2):s464.
[10] LAVIE M,DUBUISSON J. Interplay between hepatitis C virus and lipid metabolism during virus entry and assembly[J]. Biochimie,2017,141:62-69.
[11] KIM KH,HONG SP,KIM KJ,et al. HCV core protein induces hepatic lipid accumulation by activating SREBP1 and PPARγ[J].Biochem Biophys Res Commun,2007,355(4):883-888.
[12] TIAN Y,YANG Y,ZHANG X,et al. Age-dependent PPARαactivation induces hepatic sulfatide accumulation in transgenic mice carrying the hepatitis C virus core gene[J]. Glycoconj J,2016,33(6):927-936.
[13] MURTHY GD,VU K,VENUGOPAL S. Prevalence and treatment of hyperlipidemia in patients with chronic hepatitis C infection[J]. Eur J Gastroenterol Hepatol,2009,21(8):902-907.
[14] HSU CS,LIU CH,LIU CJ,et al. Association of lipid profiles with hepatitis C viral load in chronic hepatitis C patients with genotype1 or 2 infection[J]. Am J Gastroenterol,2009,104(3):598.
[15] DAI CY,CHUANG WL,HO CK,et al. Associations between hepatitis C viremia and low serum triglyceride and cholesterol levels:A community-based study[J]. J Hepatology,2008,49(1):9-16.
[16] COREY KE,KANE E,MUNROE C,et al. Hepatitis C virus infection and its clearance alter circulating lipids:Implications for long-term follow-up[J]. Hepatology,2009,50(4):1030-1037.
[17] MIRANDOLA S,BOWMAN D,HUSSAIN MM,et al. Hepatic steatosis in hepatitis C is a storage disease due to HCV interaction with microsomal triglyceride transfer protein(MTP)[J].Nutr Metab(Lond),2010,7:13.
[18] TSAI TY,WANG WT,LI HK,et al. RNA helicase DDX3 maintains lipid homeostasis through upregulation of the microsomal triglyceride transfer protein by interacting with HNF4 and SHP[J]. Sci Rep,2017,7:41452.
[19] MOGILENKO DA,DIZHE EB,SHAVVA VS,et al. Role of the nuclear receptors HNF4α,PPARα,and LXRs in the TNFα-mediated inhibition of human apolipoprotein A-I gene expression in HepG2 cells[J]. Biochemistry,2009,48(50):11950-11960.