APP基因启动子区甲基化与冠心病相关性的研究
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摘要
为了探讨淀粉样前体蛋白(amyloid precursor protein, APP)基因甲基化与冠状动脉粥样硬化性心脏病(coronary heart disease, CHD)之间的关系,采用甲基化特异性实时定量聚合酶链反应(quantitative methylation-specific PCR, qMSP)检测538名CHD患者及453名正常对照者的APP甲基化修饰水平。结果显示:CHD组的年龄、男性人数、吸烟及糖尿病患者人数均高于对照组(年龄, P=8.0E-08;男性人数, P=7.0E-07;吸烟, P=0.001;糖尿病,P=0.019)。CHD组患者白蛋白水平显著低于对照组(P=0.001),而AST、ALP及γ-GT的水平在CHD组中显著高于对照组(AST, P=3.0E-04; ALP, P=0.001;γ-GT, P=0.018)。在总体样本及男性样本中, APP基因甲基化水平在CHD组显著高于对照组(总体, P=0.026;男性, P=0.025)。在非吸烟CHD患者中, APP基因甲基化水平与狭窄程度呈正比(r=0.076, P=0.046)。在总体CHD伴高血压患者及男性CHD伴高血压患者中, APP基因甲基化水平和狭窄程度呈正比(总体, r=0.096, P=0.029;男性, r=0.135, P=0.019)。年龄分层分析后发现,在年龄≥63岁的男性CHD伴高血压患者中, APP基因甲基化水平与狭窄程度呈正比(r=0.219, P=0.020)。在年龄<63岁的不吸烟或非高血压CHD患者中, APP基因甲基化水平与狭窄程度呈负相关(不吸烟, r=-0.223, P=0.008;非高血压,r=-0.216, P=0.010)。在男性CHD患者中, APP基因甲基化水平与年龄呈正相关(r=0.163, P=0.001)。在女性CHD患者中, APP基因甲基化水平与年龄呈负相关(r=-0.192, P=0.015)。在男性正常对照组中, APP基因甲基化水平与年龄呈负相关(r=-0.203, P=0.001)。在女性正常对照组中, APP基因甲基化水平与ApoB、白蛋白及ALT水平呈负相关(r=-0.160, P=0.028; r=-0.151, P=0.036; r=-0.163, P=0.024)。在正常对照组中, APP基因甲基化水平与Lp(a)水平呈正相关(r=0.108, P=0.031)。以上结果初步表明APP基因甲基化水平增高可能和男性CHD患病风险有关。
To investigate the relationship between methylation of amyloid precursor protein(APP) gene and coronary heart disease(CHD), the APP methylation levels of 538 CHD patients and 453 normal controls were detected using the quantitative methylation-specific PCR(qMSP). The results showed that the age, the num-bers of males, smokers and diabetic patients were significantly higher in CHD patients than in controls(age,P =8.0 E-08; males, P =7.0 E-07; smokers, P =0.001; diabetes, P =0.019). Albumin levels were significantly lower in the CHD group than in the control group(P=0.001), while AST, ALP, and γ-GT levels were significantly higher in the CHD group than in control group(AST, P=3.0 E-04; ALP, P=0.001; γ-GT, P=0.018). In both total and male samples, the methylation levels of the APP gene were found to be significantly higher in the CHD group than in the control group(total samples, P=0.026; males, P=0.025). In non-smoking CHD patients, the APP methylation level was positively related to the degree of stenosis(r=0.076, P=0.046). In both total CHD patients with hypertension and CHD male patients with hypertension, APP methylation levels were found to be positively related to the degree of stenosis(total samples, r=0.096, P=0.029; males, r=0.135, P=0.019). After age stratification, it was found that the APP methylation level was positively related to stenosis in CHD male patients with hypertension aged ≥63 years(r=0.219, P=0.020). In non-smoking or non-hypertensive CHD patients aged <63 years, APP methylation levels were found to be negatively correlated with stenosis(non-smoking, r=-0.223, P=0.008; non-hypertension, r=-0.216, P=0.010). In CHD male patients, a positive correlation was found between the APP methylation level and the age(r=0.163, P=0.001). In CHD female patients, an inverse correlation was observed between the APP methylation level and the age(r=-0.192, P=0.015).In normal male controls, the APP methylation level was inversely correlated with the age(r=-0.203, P=0.001).In normal female controls, APP methylation level was inversely correlated with ApoB, albumin and ALT levels(r =-0.160, P =0.028; r =-0.151, P =0.036; r =-0.163, P =0.024, respectively). In the normal group, the APP methylation level was positively correlated with Lp(a) level(r=0.108, P=0.031). In conclusion, the above results showed that the increased methylation level of APP gene might be related to the risk of CHD in males.
To investigate the relationship between methylation of amyloid precursor protein(APP) gene and coronary heart disease(CHD), the APP methylation levels of 538 CHD patients and 453 normal controls were detected using the quantitative methylation-specific PCR(qMSP). The results showed that the age, the num-bers of males, smokers and diabetic patients were significantly higher in CHD patients than in controls(age,P =8.0 E-08; males, P =7.0 E-07; smokers, P =0.001; diabetes, P =0.019). Albumin levels were significantly lower in the CHD group than in the control group(P=0.001), while AST, ALP, and γ-GT levels were significantly higher in the CHD group than in control group(AST, P=3.0 E-04; ALP, P=0.001; γ-GT, P=0.018). In both total and male samples, the methylation levels of the APP gene were found to be significantly higher in the CHD group than in the control group(total samples, P=0.026; males, P=0.025). In non-smoking CHD patients, the APP methylation level was positively related to the degree of stenosis(r=0.076, P=0.046). In both total CHD patients with hypertension and CHD male patients with hypertension, APP methylation levels were found to be positively related to the degree of stenosis(total samples, r=0.096, P=0.029; males, r=0.135, P=0.019). After age stratification, it was found that the APP methylation level was positively related to stenosis in CHD male patients with hypertension aged ≥63 years(r=0.219, P=0.020). In non-smoking or non-hypertensive CHD patients aged <63 years, APP methylation levels were found to be negatively correlated with stenosis(non-smoking, r=-0.223, P=0.008; non-hypertension, r=-0.216, P=0.010). In CHD male patients, a positive correlation was found between the APP methylation level and the age(r=0.163, P=0.001). In CHD female patients, an inverse correlation was observed between the APP methylation level and the age(r=-0.192, P=0.015).In normal male controls, the APP methylation level was inversely correlated with the age(r=-0.203, P=0.001).In normal female controls, APP methylation level was inversely correlated with ApoB, albumin and ALT levels(r =-0.160, P =0.028; r =-0.151, P =0.036; r =-0.163, P =0.024, respectively). In the normal group, the APP methylation level was positively correlated with Lp(a) level(r=0.108, P=0.031). In conclusion, the above results showed that the increased methylation level of APP gene might be related to the risk of CHD in males.
引文
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[2]夏燕美,姬国华.早发冠心病的临床危险因素分析与护理对策[J].中华全科医学(XIA Yan-mei, JI Guo-hua. Clinical risk factor analysis and nursing strategy of premature coronary artery disease[J]. Chinese Journal of General Practice), 2017, 15(1):173-175.
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[5] IWATA A, NAGATA K, HATSUTA H, et al. Altered CpG methylation in sporadic Alzheimer’s disease is associated with APP and MAPT dysregulation[J]. Human Molecular Genetics,2014, 23(3):648-656.
[6] NICOLAS G, WALLON D, GOUPIL C, et al. Mutation in the3′untranslated region of APP as a genetic determinant of cerebral amyloid angiopathy[J]. European Journal of Human Genetics, 2016, 24(1):92-98.
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[14] D′USCIO L V, HE T, KATUSIC Z S. Expression and processing of amyloid precursor protein in vascular endothelium[J].Physiology, 2017, 32(1):20-32.
[15]王薇,齐玥.中国女性心血管病流行趋势[J].中国实用内科杂志(WANG Wei, QI Yue. Prevalence trend of cardiovascular diseases in Chinese women[J]. Chinese Journal of Practical Internal Medicine), 2014, 34(1):6-9.
[16]李威,王德征,张颖,等.天津市2010-2014年急性冠心病发病者吸烟情况分析[J].实用预防医学(LI Wei, WANG De-zheng,ZHANG Ying, et al. Smoking status of patients with acute coronary heart disease in Tianjin, 2010-2014[J]. Practical Preventive Medicine), 2018, 25(2):202-205.
[17] REGITZ-ZAGROSEK V. Sex and gender specific aspects—from cells to cardiovascular disease[M]//LEGATO M J. Principles of Gender-Specific Medicine—Gender in the Genomic Era(The Third Edition). Elsevier:Academic Press, 2017:341-362.
[18]王广娟,赵颖馨,张慧,等.吸烟及高同型半胱氨酸血症对冠心病的交互作用[J].实用医学杂志(WANG Guang-juan, ZHAO Ying-xin, ZHANG Hui, et al. Synergistic effect between smoking and hyperhomocysteinemia on incidence of coronary heart disease[J]. The Journal of Practical Medicine), 2018, 34(3):468-471.
[19]庄微,蔡晓敏,张启高.高同型半胱氨酸血症与心血管疾病相关性研究进展[J].中华实用诊断与治疗杂志(ZHUANG Wei,CAI Xiao-min, ZHANG Qi-gao. Advances in research on the relationship between hyperhomocysteinemia and cardiovascular diseases[J]. Journal of Chinese Practical Diagnosis and Therapy), 2014, 28(5):433-435.
[20]邵明学,朱建成,蔡娟,等.同型半胱氨酸与冠心病[J].现代生物医学进展(SHAO Ming-xue, ZHU Jian-cheng, CAI Juan, et al.Homocysteine and coronary heart disease[J]. Progress in Modern Biomedicine), 2013, 13(15):2983-2985.
[21] BOOTH G L, KAPRAL M K, FUNG K, et al. Relation between age and cardiovascular disease in men and women with diabetes compared with non-diabetic people:a population-based retrospective cohort study[J]. The Lancet, 2006, 368(9529):29-36.
[22] LOW WANG C C, HESS C N, HIATT W R, et al. Clinical update:cardiovascular disease in diabetes mellitus:atheroscle rotic cardiovascular disease and heart failure in type 2 diabetes mellitus-mechanisms, management, and clinical considerations[J].Circulation, 2016, 133(24):2459-2502.
[23] HATTERMANN K, MEHDORN H M, MENTLEIN R, et al. A methylation-specific and SYBR-green-based quantitative polymerase chain reaction technique for O6-methylguanine DNA methyltransferase promoter methylation analysis[J]. Analytical Biochemistry, 2008, 377(1):62-71.
[24] BASTIAN P J, ELLINGER J, WELLMANN A, et al. Diagnostic and prognostic information in prostate cancer with the help of a small set of hypermethylated gene loci[J]. Clinical Cancer Research, 2005, 11(11):4097-4106.
[25]唐军卫,黄玉晓,张鑫,等.冠心病病人危险因素年龄组分布及聚集状况[J].青岛大学医学院学报(TANG Jun-wei, HUANG Yu-xiao, ZHANG Xin, et al. Distribution and aggregation of risk factors of coronary heart disease in patients of different age groups[J]. Acta Academiae Medicinae Qingdao Universitatis), 2015, 51(3):325-328.
[2]夏燕美,姬国华.早发冠心病的临床危险因素分析与护理对策[J].中华全科医学(XIA Yan-mei, JI Guo-hua. Clinical risk factor analysis and nursing strategy of premature coronary artery disease[J]. Chinese Journal of General Practice), 2017, 15(1):173-175.
[3] UDALI S, GUARINI P, MORUZZI S, et al. Cardiovascular epigenetics:from DNA methylation to microRNAs[J]. Molecular Aspects of Medicine, 2013, 34(4):883-901.
[4] DUAN L, LIU Y, WANG J, et al. The dynamic changes of DNA methylation in primordial germ cell differentiation[J]. Gene, 2016,591(2):305-312.
[5] IWATA A, NAGATA K, HATSUTA H, et al. Altered CpG methylation in sporadic Alzheimer’s disease is associated with APP and MAPT dysregulation[J]. Human Molecular Genetics,2014, 23(3):648-656.
[6] NICOLAS G, WALLON D, GOUPIL C, et al. Mutation in the3′untranslated region of APP as a genetic determinant of cerebral amyloid angiopathy[J]. European Journal of Human Genetics, 2016, 24(1):92-98.
[7] TIBOLLA G, NORATA G D, MEDA C, et al. Increased atherosclerosis and vascular inflammation in APP transgenic mice with apolipoprotein E deficiency[J]. Atherosclerosis, 2010,210(1):78-87.
[8] VAN DE PARRE T J, GUNS P J, FRANSEN P, et al. Attenuated atherogenesis in apolipoprotein E-deficient mice lacking amyloid precursor protein[J]. Atherosclerosis, 2011, 216(1):54-58.
[9] KRISTENSEN L S, MIKESKA T, KRYPUY M, et al. Sensitive Melting Analysis after Real Time-Methylation Specific PCR(SMART-MSP):high-throughput and probe-free quantitative DNA methylation detection[J]. Nucleic Acids Research, 2008,36(7):e42
[10] MACKENZIE G, GREIG M, HAY I, et al. Competing risk analysis of factors related to long-term incidence of CHD[J].Journal of Epidemiology and Community Health, 2017, 71(1):33-36.
[11] TEODORIDIS J M, STRATHDEE G, BROWN R. Epigenetic silencing mediated by CpG island methylation:potential as a therapeutic target and as a biomarker[J]. Drug Resistance Updates, 2004, 7(4-5):267-278.
[12] FUKUMOTO R, KAWAI M, MINAI K, et al. Conflicting relationship between age-dependent disorders, valvular heart disease and coronary artery disease by covariance structure analysis:possible contribution of natriuretic peptide[J]. PLoS One,2017, 12(7):e0181206.
[13] ABBOTT R D, CURB J D, RODRIGUEZ B L, et al. Age-related changes in risk factor effects on the incidence of coronary heart disease[J]. Annals of Epidemiology, 2002, 12(3):173-181.
[14] D′USCIO L V, HE T, KATUSIC Z S. Expression and processing of amyloid precursor protein in vascular endothelium[J].Physiology, 2017, 32(1):20-32.
[15]王薇,齐玥.中国女性心血管病流行趋势[J].中国实用内科杂志(WANG Wei, QI Yue. Prevalence trend of cardiovascular diseases in Chinese women[J]. Chinese Journal of Practical Internal Medicine), 2014, 34(1):6-9.
[16]李威,王德征,张颖,等.天津市2010-2014年急性冠心病发病者吸烟情况分析[J].实用预防医学(LI Wei, WANG De-zheng,ZHANG Ying, et al. Smoking status of patients with acute coronary heart disease in Tianjin, 2010-2014[J]. Practical Preventive Medicine), 2018, 25(2):202-205.
[17] REGITZ-ZAGROSEK V. Sex and gender specific aspects—from cells to cardiovascular disease[M]//LEGATO M J. Principles of Gender-Specific Medicine—Gender in the Genomic Era(The Third Edition). Elsevier:Academic Press, 2017:341-362.
[18]王广娟,赵颖馨,张慧,等.吸烟及高同型半胱氨酸血症对冠心病的交互作用[J].实用医学杂志(WANG Guang-juan, ZHAO Ying-xin, ZHANG Hui, et al. Synergistic effect between smoking and hyperhomocysteinemia on incidence of coronary heart disease[J]. The Journal of Practical Medicine), 2018, 34(3):468-471.
[19]庄微,蔡晓敏,张启高.高同型半胱氨酸血症与心血管疾病相关性研究进展[J].中华实用诊断与治疗杂志(ZHUANG Wei,CAI Xiao-min, ZHANG Qi-gao. Advances in research on the relationship between hyperhomocysteinemia and cardiovascular diseases[J]. Journal of Chinese Practical Diagnosis and Therapy), 2014, 28(5):433-435.
[20]邵明学,朱建成,蔡娟,等.同型半胱氨酸与冠心病[J].现代生物医学进展(SHAO Ming-xue, ZHU Jian-cheng, CAI Juan, et al.Homocysteine and coronary heart disease[J]. Progress in Modern Biomedicine), 2013, 13(15):2983-2985.
[21] BOOTH G L, KAPRAL M K, FUNG K, et al. Relation between age and cardiovascular disease in men and women with diabetes compared with non-diabetic people:a population-based retrospective cohort study[J]. The Lancet, 2006, 368(9529):29-36.
[22] LOW WANG C C, HESS C N, HIATT W R, et al. Clinical update:cardiovascular disease in diabetes mellitus:atheroscle rotic cardiovascular disease and heart failure in type 2 diabetes mellitus-mechanisms, management, and clinical considerations[J].Circulation, 2016, 133(24):2459-2502.
[23] HATTERMANN K, MEHDORN H M, MENTLEIN R, et al. A methylation-specific and SYBR-green-based quantitative polymerase chain reaction technique for O6-methylguanine DNA methyltransferase promoter methylation analysis[J]. Analytical Biochemistry, 2008, 377(1):62-71.
[24] BASTIAN P J, ELLINGER J, WELLMANN A, et al. Diagnostic and prognostic information in prostate cancer with the help of a small set of hypermethylated gene loci[J]. Clinical Cancer Research, 2005, 11(11):4097-4106.
[25]唐军卫,黄玉晓,张鑫,等.冠心病病人危险因素年龄组分布及聚集状况[J].青岛大学医学院学报(TANG Jun-wei, HUANG Yu-xiao, ZHANG Xin, et al. Distribution and aggregation of risk factors of coronary heart disease in patients of different age groups[J]. Acta Academiae Medicinae Qingdao Universitatis), 2015, 51(3):325-328.