1999 년도산학협동연구비 Korean Circulation J 2001;315:512-519 한국인에서 Endothelial Nitric Oxide Synthase(eNOS) 유전자변이빈도및관동맥연축과의상관관계 김정희 2 최성준 1 김보영 2 이정우 2 강정아 2 임대승 2 이민수 2 김윤철 2 성보영 2 임병관 1 성인환 2 전은석 1 Incidence of enos Gene Mutation in Korean Patients with Coronary Artery Spasm Jung-Hee Kim, MD 2, Seong-Choon Choe, MD 1, Bo-Young Kim, MD 2, Jeong-Woo Lee, MD 2, Jung-A Kang, MD 2, Dae-Seung Lim, MD 2, Min-Soo Lee, MD 2, Yoon-Cheol Kim, MD 2, Bo-Young Sung, MD 2, Byung-Kwan Lim, MS 1, In-Whan Seong, MD 2 and Eun-Seok Jeon, MD 1 1 Department of Internal Medicine, Soonchunhyang University College of Medicine, Bucheon, 2 Department of Internal Medicine, Chungnam National University College of Medicine, Taejeon, Korea ABSTRACT BackgroundCoronary artery spasm is an important mechanism in producing myocardial ischemia. But the exact mechanism of the spasm is not well known. We investigated the mutation of endothelial nitric oxide synthase enos that produce nitric oxide and relationship between enos mutation and coronary artery spasm. Materials and MethodsBlood were drawn from the patients with angiographically proven coronary artery spasm and normal controls. DNA were extracted and polymerase chain reaction and restriction analysis with Nae I were performed to find T 786 C mutation of 5 -flanking region in enos gene. Clinical and lipid parameters were reviewed. ResultsThe prevalence of coronary artery spasm in Chungnam National University Hospital was 19% 125/664. The 85 cases of coronary artery spasm and 131 cases of normal control were compared. The men in spasm group were more prevalent than the women p0.0001 and rate of smoking were higher in spasm group p0.0001. The T 786 C mutation of 5 -flanking region were 20% in spasm group and 3% in normal control p0.0001. There were no differences in clinial profiles between mutant group and non-mutant group. But total cholesterol p0.04, triglyceride p0.006, LDL-cholesterol p0.02 in mutant group were higher than non-mutant group. ConclusionThe T 786 C mutation of 5 -flanking region in enos gene was related to coronary artery spasm and lipid profiles in Korea. Korean Circulation J 2001;305:512-519 KEY WORDSEndothelial nitric oxide synthase Nitric oxide Coronary artery spasm Mutation Lipid. 서 론 512
대상및방법 환자군및대조군선택 혈액채취 513
Genomic DNA(gDNA) 추출 enos 유전자증폭시발체 (primer) 제작 중합효소연쇄반응과중합효소연쇄반응산물에대한제한분석 (Restriction analysis) 통계적분석 결과 대상 enos gene 변이의빈도 514 Korean Circulation J 2001;313:512-519
형협심증 환자에서 돌연변이가 관찰된 군과 관찰되지 않은 군의 임상적 특징에는 큰 차이가 없었고, 남녀간 고 찰 의 차이도 없었다. 그러나 돌연변이가 있었던 군에서 총 콜레스테롤치(p 0.04), 중성지방치(p 0.006), 저밀도 한 병원에서 3년간 관동맥조영술의 19%에서 관동맥 지단백 콜레스테롤치(p 0.02)가 통계적으로 유의하게 연축이 보고되었다는 것은 상당히 의미 있는 보고로 생 높게 나타났다. 고밀도 지단백 콜레스테롤치는 통계적 각이 되는데 그 이유는 관동맥 연축을 의심하지 않고는 으로 차이가 없었다(Table 3 and 4). 유발검사를 하지 못하기 때문에 실제로 우리 나라에서 Fig. 1. Schematic diagram which demonstrated endothelial nitric oxide (enos) gene and Nae I restriction site. Fig. 2. Electrophoretic photograph of polymerase chain reaction (PCR) and restriction anlaysis by Nae I restriction enzyme (Lane 1, 3, 5, 7, 9 PCR product before Nae I restriction, Lane 2, 4, 6, 8, 10 PCR product after Nae I restriction). Lane 2, 6 were restriction fragment length polymorphism (RFLP) positive and lane 4, 8, 10 were RFLP negative. Fig. 3. Sequence of the amplified polymerase chain reaction (PCR) product with Nae I restiction sitepositive sample and -negative sample. Base thymine (T) was replaced to cytosine (C) and Nae I restriction site was appeared. Thus the PCR product was cleaved. 515
Table 1. Difference of clinical profiles between coronary artery spasm group and spasm-negative group Spasm Spasm p Numbers 85 131 Sex M/F 74/11 81/50 0.0001 Age yr 5810 5512 0.13 Diabetes 9 11% 18 14% 0.64 Hypertension 31 36% 33 25% 0.11 Obesity 11 13% 31 24% 0.08 Family history 3 4% 4 3% 0.84 Smoking 53 62% 27 21% 0.0001 ObesityBody mass index25 Table 2. Difference of lipid profiles and RFLP between coronary artery spasm group and spasm-negative group Unitmg/dl Spasm Spasm p Numbers 85 131 Triglyceride 16794 169156 0.93 Total cholesterol 18544 188 43 0.62 HDL-C 4410 47 17 0.09 LDL-C 11139 113 45 0.75 Nae I RFLP 17 20% 4 3% 0.0001 HDL-Chigh density lipoprotein-cholesterol LDL-Clow density lipoprotein-cholesterol RFLPrestriction fragment length polymorphism Table 3. Differnce of clinical profiles between mutant group and mutant-negative group in variant angina mutant grouprflp, mutant-negative group RFLP RFLP RFLP p Numbers 17 68 Sex M/F 15/2 59/9 0.81 Age yr 5610 5810 0.51 Diabetes 1 6% 8 12% 0.79 Hypertension 7 41% 24 35% 0.87 Obesity 3 18% 8 12% 0.81 Family history 2 12% 1 1% 0.19 Smoking 10 59% 43 63% 0.96 ObesityBody mass index25 RFLPrestriction fragment length polymorphism Table 4. Differnce of lipid profiles between mutant group and mutant-negative group in variant angina mutant grouprflp, mutant-negative group RFLP Unitmg/dl Spasm Spasm p Numbers 17 68 Triglyceride 214112 15687 0.04 Total cholesterol 214 42 17538 0.006 HDL-C 42 7 4411 0.66 LDL-C 134 45 10635 0.02 HDL-Chigh density lipoprotein-cholesterol LDL-Clow density lipoprotein-cholesterol RFLPrestriction fragment length polymorphism Table 5. Incidence of T 786 C mutation in Japan and Korea In Japan Controls Variant /homozygote 0% 2% /heterozygote 7% 28% /homozygote 93% 70% In Korea /homozygote 0% 0% /heterozygote 3% 20% /homozygote 97% 80% 516 Korean Circulation J 2001;313:512-519
요약 연구목적 : 대상및방법 : 517
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