血管紧张素转换酶基因多态性（I/D）与慢性阻塞性肺病、肺心病的关系分析
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摘要
近来，大量国内外研究证明肾素—血管紧张素—醛固酮(RAS)与慢性阻塞性肺病(COPD)，肺心病，肺通气障碍，肺动脉高压等有关。在RAS中血管紧张素转换酶(ACE)是一关键酶。ACE是肾素—血管紧张素—醛固酮系统以及缓激肽系统的重要调节因素，影响人体多种生理机能。由于肺毛细血管内皮细胞是血中ACE的主要来源，长期以来人们以期利用血液中ACE改变来诊断肺部疾病，但在各种肺部疾病中血液ACE活性改变报告不一。近年来ACE基因作为研究各类疾病的遗传易感性的“候选基因”而受到人们的关注。ACE在16内含子存在287bp插入片段及缺失片段，呈现ACE基因插入/缺失多态性，在人类存在D、I两种等位基因，组成DD型(纯合子)II(纯合子) ID (杂合子)三种基因类型。ACE基因插入/缺失多态性与血清ACE水平有显著相关性，DD型血清ACE水平最高，II型血清ACE水平最低，杂合子ID型的介于两则之间(1)。本课题拟探明ACE 活性及其基因多态性与COPD，肺心病的相关性。
方法1、设立健康人对照组、慢性阻塞性肺部疾病(COPD)组、肺心病组各30例。2、用小剂量全血DNA抽提试剂盒提取白细胞DNA。3、
用多聚酶链反应(PCR)检测各组中的ACE基因I/D多态性(分别有三种基因型DD、ID、II)。4、用紫外分光光度法检测血清中的ACE(sACE)，
结果 1、在ACE基因多态性中，DD型的 sACE水平最高，II型的sACE水平最低，ID基因型的介于两者之间，DD、ID、II的sACE值分别为565.02±128.79u/l、516.30±117.01u/l、 439.13±105.18u/l。2、COPD及肺心病中sACE值明显高于健康人组，存在显著性差异(P<0.01) 3、COPD及肺心病组与健康人组的I/D等位基因比较有显著性差异(x2=6.599,P=0.01)。COPD组、肺心病组以D等位基因占优势，而健康人对照组以I等位基因占优势。4、COPD组、肺心病组中等位基因D与I的优势比(OR)均为2.6。结论 1、 ACE基因多态性与sACE有相关性。2、COPD及肺心病中sACE增高可能是携带D等位基因的个体高表达ACE。3、ACE 的D等位基因可能是患COPD及肺心病的危险因素。
Recent studies have shown that the renin-angiotensin system (RAS) correlates with diseases of respiratory system, such as chronic obstructive pulmonary disease (COPD) and pulmonary heart disease. Angiotensin-converting enzyme (ACE) is a key enzyme in RAS, which plays an important role in the RAS and bradykinin system and affects a variety of physiological function in human body. As circulating ACE mainly originates from the pulmonary capillary endothelial cells, it has been hoped to diagnose lung diseases by measuring ACE levels, but reports about ACE levels in lung diseases are varied. Recent interests considered the ACE gene as a candidate sensitive gene to research relative diseases. The ACE gene contains a polymorphism based on the presence (insertion [1]) or absence (deletion [D]) in three genotypes (DD and II homozygotes and ID heterozygotes). The polymorphism of ACE is significantly correlated with
serum ACE (sACE). The sACE levels in DD individuals have been found to be the highest of those in the ACE three genotypes, while in II individuals is the lowest and ID individuals is in between. The aim of this work was to investigate the relationship of polymorphism of ACE gene with COPD and pulmonary heart disease.
Method: ⑴ 30 healthy subjects, 30 patients with COPD, 30 patients with pulmonary heart disease were studied. ⑵ DNA of white blood cells was abstracted followed by instruction of the reagent box drawing DNA of white blood cells from small dose of whole blood. ⑶ ACE genotypes (DD, ID, II genotypes respectively) were determined in all subjects by polymerase chain reaction (PCR). Meanwhile, sACE levels was determined in all subjects by ultraviolet spectrophotometry.
Result: ⑴ sACE level in DD individuals had been found to be the highest of those in ACE three genotypes, while in II individuals was the lowest and in II individuals in between. The sACE levels were (565.02±128.79), (516.30±117.01) and (439.13±105.18)(/L, respectively for genotypes DD,ID and II. ⑵ The levels of sACE in patients with COPD or pulmonary heart disease were significantly higher than in healthy subjects (P<0.01). ⑶ The percentage of I/D allele in patients with COPD and pulmonary heart disease was significantly differed form healthy subjects
(x2=6.599, P=0.01). ⑷ The odds ratio of patients (COPD and pulmonary heart disease) with D allele and those with I allele was both 2.6.
Conclusion: ⑴ There was a close relation between the sACE levels and ACE genotypes of DD, ID and II. ⑵ The high expression of ACE is due to D allele. (3)The results suggested that D allele of ACE might be a risk factor for COPD and pulmonary heart disease.
方法1、设立健康人对照组、慢性阻塞性肺部疾病(COPD)组、肺心病组各30例。2、用小剂量全血DNA抽提试剂盒提取白细胞DNA。3、
用多聚酶链反应(PCR)检测各组中的ACE基因I/D多态性(分别有三种基因型DD、ID、II)。4、用紫外分光光度法检测血清中的ACE(sACE)，
结果 1、在ACE基因多态性中，DD型的 sACE水平最高，II型的sACE水平最低，ID基因型的介于两者之间，DD、ID、II的sACE值分别为565.02±128.79u/l、516.30±117.01u/l、 439.13±105.18u/l。2、COPD及肺心病中sACE值明显高于健康人组，存在显著性差异(P<0.01) 3、COPD及肺心病组与健康人组的I/D等位基因比较有显著性差异(x2=6.599,P=0.01)。COPD组、肺心病组以D等位基因占优势，而健康人对照组以I等位基因占优势。4、COPD组、肺心病组中等位基因D与I的优势比(OR)均为2.6。结论 1、 ACE基因多态性与sACE有相关性。2、COPD及肺心病中sACE增高可能是携带D等位基因的个体高表达ACE。3、ACE 的D等位基因可能是患COPD及肺心病的危险因素。
Recent studies have shown that the renin-angiotensin system (RAS) correlates with diseases of respiratory system, such as chronic obstructive pulmonary disease (COPD) and pulmonary heart disease. Angiotensin-converting enzyme (ACE) is a key enzyme in RAS, which plays an important role in the RAS and bradykinin system and affects a variety of physiological function in human body. As circulating ACE mainly originates from the pulmonary capillary endothelial cells, it has been hoped to diagnose lung diseases by measuring ACE levels, but reports about ACE levels in lung diseases are varied. Recent interests considered the ACE gene as a candidate sensitive gene to research relative diseases. The ACE gene contains a polymorphism based on the presence (insertion [1]) or absence (deletion [D]) in three genotypes (DD and II homozygotes and ID heterozygotes). The polymorphism of ACE is significantly correlated with
serum ACE (sACE). The sACE levels in DD individuals have been found to be the highest of those in the ACE three genotypes, while in II individuals is the lowest and ID individuals is in between. The aim of this work was to investigate the relationship of polymorphism of ACE gene with COPD and pulmonary heart disease.
Method: ⑴ 30 healthy subjects, 30 patients with COPD, 30 patients with pulmonary heart disease were studied. ⑵ DNA of white blood cells was abstracted followed by instruction of the reagent box drawing DNA of white blood cells from small dose of whole blood. ⑶ ACE genotypes (DD, ID, II genotypes respectively) were determined in all subjects by polymerase chain reaction (PCR). Meanwhile, sACE levels was determined in all subjects by ultraviolet spectrophotometry.
Result: ⑴ sACE level in DD individuals had been found to be the highest of those in ACE three genotypes, while in II individuals was the lowest and in II individuals in between. The sACE levels were (565.02±128.79), (516.30±117.01) and (439.13±105.18)(/L, respectively for genotypes DD,ID and II. ⑵ The levels of sACE in patients with COPD or pulmonary heart disease were significantly higher than in healthy subjects (P<0.01). ⑶ The percentage of I/D allele in patients with COPD and pulmonary heart disease was significantly differed form healthy subjects
(x2=6.599, P=0.01). ⑷ The odds ratio of patients (COPD and pulmonary heart disease) with D allele and those with I allele was both 2.6.
Conclusion: ⑴ There was a close relation between the sACE levels and ACE genotypes of DD, ID and II. ⑵ The high expression of ACE is due to D allele. (3)The results suggested that D allele of ACE might be a risk factor for COPD and pulmonary heart disease.
引文
1．Brigitte R,Christine H,Francois A,et al.An Insertion/Deletion Polymorphism in the Angiotensin I-converting Enzyme Gene Accounting for Half the Variance of Serum Enzyme Levels. J Clin Invest 1990,86:1343-1346.
2．Alhenc-Gelas FJ,Richard D Courbon J.Distribution of plasma angiotensin I-converting enzyme in healthy men.Relationship to enviromental and hormonal parameter J Clin lab ,ed In Press.
3．Jan D,Schalekamp M,Bax W,et al.ACE in the human heart:effect of the deletion/insertion polymorphism.Circulation 1995;92:1387-1388.
4．Tiret L, Rigat B,Visvikis S, et al.Evidence from combined segregation and linkage analysis,that a variant of the angiotensin I-converting gene controls plasma ACE levels.Am J Hum Genet 1992;51:197-205.
5．Rigat B,Hubert C,Corvol P,et al.PCR detection of the insertion/deletion polymorphism of the human angiotensin converting enzyme gene(DCPI)(dipeptidyl carboxypeptidase 1)Nucleic Acid Res,1992,20:1433.
6．项坤三，王延庆，吴松华。血管紧张素I转移酶基因与中国人冠心病、糖尿病及高血压的关系。中华内分泌代谢杂志，1995，11：201。
7．王洪武，聂舟山，段蕴铀等。肺癌及慢性阻塞性肺疾病患者的血管紧张素转移酶基因多态性的研究。北京医学，2000，22(6)：364-365。
8．Millar EA, Angus RM, Hulks G, et al. Activity of the renin-angiotensin system in acute severe asthma and the effect of angiotensin II on lung function, Thrict, 1994,49:492-495.
9．Yamawaki I, Tamaoki J, Horii S, et al. Angiotensin II potentiates neurally mediated contraction of rabbit airway smooth muscles. Am Rev Resir dis, 1991,143:A343.
10．Kato H, Suzuki H, Tajima S, Ogata Y, et al. Angiotensin II stimulates collagen synthesis in cultures vascular smooth muscle cells. J Hyoertens 1991;9:17-22.
11．Imai T,Hirata Y,Emorn T,Yanagisawa M,et al.Induction of endothelin-1 gene by angiotensin and vasioressin in endothelial cells.Hypertension 1992;19:753.
12．Robert J,Van S,Emiel F,et al. The DD genotype of the angiotensin converting enzyme gene negatively associated with right ventricular hypertrophy in male patients with chronic obstructive pulmonary disease.Am J Respir Crit Care M ed ,1999,159:1791-1795.
13．Abraham WT,Raynolds MV, Gottschall B,et al. Importance of angiotensin converting enzyme in pulmonary hypertension. Cardiology .1995,86(Suppl.1):9-15.
14．Hiroshi K,Talashi O,Kazuto H, et al.Deletion polymorphisms in the angiotensin converting enzyme gene are associated with pulmonary hyopertension evoked by exercise challenge in patients with chronic obstructive pulmonary disease.Am J Respir Crit Care Med,2000;162:1235-1238.
15．Ucer G Yeldirim Z, Ataol E, et al.Serum angiotensin converting enzyme activity in pulmonary diseases:correlation with lung function parameters.Life Sci, 1997,61(11):1075-1082.
16．Hollinger MA,Giri SN,Patwell S.et al.Effect of acute lung injury on angiotensin converting enzyme in serum,lung lavage and effusate.Am Rev Respir Dis 1980;121:373.
17．Hubert C,Houot AM,Corvol P. et al.Structure of the angiotensin I-converting enzyme gene:two alternate promoters correspond to evolutionary steps of a duplicated gene,J Biol Chem 1991;266:15377-15383.
18．王琳，冯华松。肺心病患者肾素系统活性和心钠素及其与病情的关系。陕西医学杂志。1993；22(10)：569-582。
19．Lydie H,Mud R,Amma V,et al.Association of 3 gene polymorphisms weth atopic diseases. J Allergy Clin Immunol 1998;103(4):702-707.
20．柳青、雷寒、王曦。血管紧张素转移酶基因型与重庆地区人群原发性高血压药物治疗的关系。中华医学遗传学杂志，2000，17(5)；340-341。
21．李颖、袁方、刘青。肺心病患者血管紧张素转换酶测定的临床观察[J]。现代中西医结合杂志1999；8(11)：1766-1767。
2．Alhenc-Gelas FJ,Richard D Courbon J.Distribution of plasma angiotensin I-converting enzyme in healthy men.Relationship to enviromental and hormonal parameter J Clin lab ,ed In Press.
3．Jan D,Schalekamp M,Bax W,et al.ACE in the human heart:effect of the deletion/insertion polymorphism.Circulation 1995;92:1387-1388.
4．Tiret L, Rigat B,Visvikis S, et al.Evidence from combined segregation and linkage analysis,that a variant of the angiotensin I-converting gene controls plasma ACE levels.Am J Hum Genet 1992;51:197-205.
5．Rigat B,Hubert C,Corvol P,et al.PCR detection of the insertion/deletion polymorphism of the human angiotensin converting enzyme gene(DCPI)(dipeptidyl carboxypeptidase 1)Nucleic Acid Res,1992,20:1433.
6．项坤三，王延庆，吴松华。血管紧张素I转移酶基因与中国人冠心病、糖尿病及高血压的关系。中华内分泌代谢杂志，1995，11：201。
7．王洪武，聂舟山，段蕴铀等。肺癌及慢性阻塞性肺疾病患者的血管紧张素转移酶基因多态性的研究。北京医学，2000，22(6)：364-365。
8．Millar EA, Angus RM, Hulks G, et al. Activity of the renin-angiotensin system in acute severe asthma and the effect of angiotensin II on lung function, Thrict, 1994,49:492-495.
9．Yamawaki I, Tamaoki J, Horii S, et al. Angiotensin II potentiates neurally mediated contraction of rabbit airway smooth muscles. Am Rev Resir dis, 1991,143:A343.
10．Kato H, Suzuki H, Tajima S, Ogata Y, et al. Angiotensin II stimulates collagen synthesis in cultures vascular smooth muscle cells. J Hyoertens 1991;9:17-22.
11．Imai T,Hirata Y,Emorn T,Yanagisawa M,et al.Induction of endothelin-1 gene by angiotensin and vasioressin in endothelial cells.Hypertension 1992;19:753.
12．Robert J,Van S,Emiel F,et al. The DD genotype of the angiotensin converting enzyme gene negatively associated with right ventricular hypertrophy in male patients with chronic obstructive pulmonary disease.Am J Respir Crit Care M ed ,1999,159:1791-1795.
13．Abraham WT,Raynolds MV, Gottschall B,et al. Importance of angiotensin converting enzyme in pulmonary hypertension. Cardiology .1995,86(Suppl.1):9-15.
14．Hiroshi K,Talashi O,Kazuto H, et al.Deletion polymorphisms in the angiotensin converting enzyme gene are associated with pulmonary hyopertension evoked by exercise challenge in patients with chronic obstructive pulmonary disease.Am J Respir Crit Care Med,2000;162:1235-1238.
15．Ucer G Yeldirim Z, Ataol E, et al.Serum angiotensin converting enzyme activity in pulmonary diseases:correlation with lung function parameters.Life Sci, 1997,61(11):1075-1082.
16．Hollinger MA,Giri SN,Patwell S.et al.Effect of acute lung injury on angiotensin converting enzyme in serum,lung lavage and effusate.Am Rev Respir Dis 1980;121:373.
17．Hubert C,Houot AM,Corvol P. et al.Structure of the angiotensin I-converting enzyme gene:two alternate promoters correspond to evolutionary steps of a duplicated gene,J Biol Chem 1991;266:15377-15383.
18．王琳，冯华松。肺心病患者肾素系统活性和心钠素及其与病情的关系。陕西医学杂志。1993；22(10)：569-582。
19．Lydie H,Mud R,Amma V,et al.Association of 3 gene polymorphisms weth atopic diseases. J Allergy Clin Immunol 1998;103(4):702-707.
20．柳青、雷寒、王曦。血管紧张素转移酶基因型与重庆地区人群原发性高血压药物治疗的关系。中华医学遗传学杂志，2000，17(5)；340-341。
21．李颖、袁方、刘青。肺心病患者血管紧张素转换酶测定的临床观察[J]。现代中西医结合杂志1999；8(11)：1766-1767。