运动后血清肌酸激酶异常增高的现象与ACE/CK-MM基因多态性研究
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摘要
众所周知,在高强度运动后可出现肌痛、肌酸激酶增高等横纹肌溶解的症状,甚至导致严重并发症。近来人们逐渐发现,部分健康人群在中低强度下就出现血清CK的异常增高,其增高幅度与运动负荷程度不成比例,有学者将其称为运动后血清肌酸激酶“高反应者”(high responders,HR)。在排除体格、身体素质、训练状况、身体脂肪含量以及某些酶的缺乏后,人们考虑这种现象与基因差异有关。近几年来我们在临床中也发现数例年轻士兵为HR,在中低强度负荷训练后,以乏力、黑尿等症状就诊,伴血清CK明显增高,个别患者还反复发作;还发现一对健康父子同为HR,故推测HR与基因有关,并通过对部分HR的基因分析,已发现两个可能与之有关的耐力基因。为了查明HR在中国汉族中的分布特点并探讨其与ACE基因I/D多态性及CKMM基因限制性酶切片段多态性的关系,我们筛选出201名健康新兵进行3000m跑,对肌酸激酶、乳酸脱氢酶、碱性磷酸酶、转氨酶及电解质进行检测,通过百分位点初步确定HR界定值,然后采用PCR及限制性酶切多态性分析技术,对所有受试者ACE、CKMM基因多态性进行检测、分析。结果发现,在我们的运动强度下HR的CK值为577IU/L,有20名受试者为高反应者。中国汉族正常对照组与高加索正常对照组在ACE/CK-MM基因多态性方面均有明显差异(P＜0.05)。中国汉族HR的ACE基因频率I=53%,D=48%,较对照组D等位基因频率高,但无统计学意义(P=0.06);DD基因型(25%)也明显高于对照组的基因型分布,无统计学意义(P=0.06);无论DD基因型还是D等位基因与高加索正常对照人群之间的差异无统计学意义(P=0.10,P=0.29)。中国汉族HR CK-MM基因频率A=65%,G-35%,较汉族对照组G等位基因频率高(P＜0.05);GG基因型也明显高于汉族对照组(P＜0.05);无论基因型还是基因频率与高加索正常人群对照相比较均无统计学差异(P＞0.05)。结论:1.普通人群中约有10%左右个体运动后易出现肌酸激酶高反应,在本研究中高反应者肌酸激酶值为577 IU/L。2.ACE I/D基因多态性与运动后肌酸激酶高反应有关,D等位基因较I等位基因易出现高反应。本研究的高反应者基因分布接近于高加索正常人群。3.CK-MM A/G基因多态性与运动后肌酸激酶高反应明显相关,G等位基因更易出现高反应。本研究的高反应者基因分布接近于高加索正常人群。4.同时具有GG、DD基因型个体更易出现运动后肌酸激酶异常升高,D+G基因频率有相加作用。
It was well known that high intensity exercise may lead to myalgia, increased creatine kinase(CK) and other symptoms of rhabdomyolysis,and even result in serious complications.However,some healthy people had CK markedly elevated after moderate or mild intensity exercise and their proportional was asymmetric,some scholars named these people,that CK obviously elevation post moderate or mild intensity exercise,as high responders(HR).After excluded the medical,fitness,training status,body fat content,and the lack of some enzymes,people considered that genetic differences made the phenomenon.In recent years our department also diagnosed several young soldiers as HR,which with fatigue,dark urinary and significantly increased serum CK after low-intensity training,some individuals recurrent episodes.A man and his son both were diagnosed as HR, so the HR related with gene.We thought two genes may related with HR after analyzed the gene of some HR.In order to find out distribution characteristic of HR in the Han population of China and to study the their relation with the ACE gene I/D polymorphism and restriction fragment length polymorphism(RFLP) in the CK-MM gene,we screened 201 healthy recruits to run 3000m and tested some the creatine kinase and other biochemical index.We defined the CK value of HR by centile of biochemical index,then detected and analyzed ACE/CKMM gene polymorphism by PCR and RFLP technique.The results showed that the CK value of HR was 577 IU/L,there are 20 subjects were the high responders.Between Han control and Caucasian control,the allele frequencies and genotype of ACE/CK-MM gene polymorphisms had significant difference(P＜0.05).the ACE gene frequency of HR in Han as follows:I=53%,D=48%,the D allele frequency was higher than Han control group,but no statistical significance(P=0.06); DD genotype(25%) also was significantly higher than that of control without statistical difference(P=0.06);Between the Caucasus group and Han HR, DD genotype and D allele were no statistical difference(P=0.10,P=0.29). CK-MM gene frequency of Han HR as follows:A=65%,G=35%.G allele frequency of HR was higher than that of Han control(P＜0.05);GG genotype also obviously higher than that of Han control(P＜0.05);The allele frequency and genotype of CK-MM in HR of Han population were no significant difference from that of Caucasian(P＞0.05),compared to the Han controls were significant difference with Caucasian(p＜0.05).Conclusion:1.To general population,about 10%of individuals prone to abnormal CK elevated post exercise,and the threshold of HR was 577IU/L in this study.2.ACE D allele related with HR.In this study,the ACE genotype distribution of the HR closed to the Caucasus control.3.CK-MM G gene had obviously correlated with HR.The CK-MM genotype distribution of the HR closed to the Caucasus control.4.The individual with GG/DD genotype was more vulnerable to CK abnormal increase post-exercise,D and G allele had a additive action to muscle injury after exercise.
It was well known that high intensity exercise may lead to myalgia, increased creatine kinase(CK) and other symptoms of rhabdomyolysis,and even result in serious complications.However,some healthy people had CK markedly elevated after moderate or mild intensity exercise and their proportional was asymmetric,some scholars named these people,that CK obviously elevation post moderate or mild intensity exercise,as high responders(HR).After excluded the medical,fitness,training status,body fat content,and the lack of some enzymes,people considered that genetic differences made the phenomenon.In recent years our department also diagnosed several young soldiers as HR,which with fatigue,dark urinary and significantly increased serum CK after low-intensity training,some individuals recurrent episodes.A man and his son both were diagnosed as HR, so the HR related with gene.We thought two genes may related with HR after analyzed the gene of some HR.In order to find out distribution characteristic of HR in the Han population of China and to study the their relation with the ACE gene I/D polymorphism and restriction fragment length polymorphism(RFLP) in the CK-MM gene,we screened 201 healthy recruits to run 3000m and tested some the creatine kinase and other biochemical index.We defined the CK value of HR by centile of biochemical index,then detected and analyzed ACE/CKMM gene polymorphism by PCR and RFLP technique.The results showed that the CK value of HR was 577 IU/L,there are 20 subjects were the high responders.Between Han control and Caucasian control,the allele frequencies and genotype of ACE/CK-MM gene polymorphisms had significant difference(P＜0.05).the ACE gene frequency of HR in Han as follows:I=53%,D=48%,the D allele frequency was higher than Han control group,but no statistical significance(P=0.06); DD genotype(25%) also was significantly higher than that of control without statistical difference(P=0.06);Between the Caucasus group and Han HR, DD genotype and D allele were no statistical difference(P=0.10,P=0.29). CK-MM gene frequency of Han HR as follows:A=65%,G=35%.G allele frequency of HR was higher than that of Han control(P＜0.05);GG genotype also obviously higher than that of Han control(P＜0.05);The allele frequency and genotype of CK-MM in HR of Han population were no significant difference from that of Caucasian(P＞0.05),compared to the Han controls were significant difference with Caucasian(p＜0.05).Conclusion:1.To general population,about 10%of individuals prone to abnormal CK elevated post exercise,and the threshold of HR was 577IU/L in this study.2.ACE D allele related with HR.In this study,the ACE genotype distribution of the HR closed to the Caucasus control.3.CK-MM G gene had obviously correlated with HR.The CK-MM genotype distribution of the HR closed to the Caucasus control.4.The individual with GG/DD genotype was more vulnerable to CK abnormal increase post-exercise,D and G allele had a additive action to muscle injury after exercise.
引文
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1.Lin H,Chie W,Lien H.Epidemiological analysis of factors influencing an episode of exertional rhabdomyolysis in high school students.Am J Sports Med,2006,34:481-486.
2.Moeller J and McKeag D.Exercise-induced rhabdomyolysis.Sports Med Arthrosc Rev,2004,3:274-279.
3.Bagley WH,Yang H,Shah KH.Rhabdomyolysis.Intern Emerg Med,2007,2:210-8.
4.Hurley JK.Severe rhabdomyolysis in well conditioned athletes.Mil Med,1989,154:244-5.
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6.Roth SM,Martel GF,Rogers MA.Muscle biopsy and muscle fiber hypercontraction:a brief review.Eur J Appl Physiol,2000,83:239-45.
7.Visweswaran P,Guntupalli J.Rhabdomyolysis.Crit Care Clin,1999,15:415-28.
8.Clarkson PM,Hubal MJ.Exercise-induced muscle damage in humans.Am J Phys Med Rehabil,2002,81:S52-69.
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19.Heled Y,Bloom MS,Wu TJ,et al.CK-MM and ACE genotypes and physiological prediction of the creatine kinase response to exercise.J Appl Physiol,2007,103:504-10.
20.Kauwe JS,Wang J,Mayo K,et al.Alzheimer's disease risk variants show association with cerebrospinal fluid amyloid beta.Neurogenetics,2009,10:13-7.
21.Osawa N,Koya D,Araki S,et al.Combinational effect of genes for the renin-angiotensin system in conferring susceptibility to diabetic nephropathy.J Hum Genet,2007,52:143-51.
22.Zakrzewski-Jakubiak M,de Denus S.Ten renin-angiotensin system-related gene polymorphisms in maximally treated Canadian Caucasian patients with heart failure.Br J Clin Pharmacol.2008,65:742-51.
23.Myerson S,Hemingway H,Budget R,et al.Human angiotensin I converting enzyme gene and endurance performance.J Appl Physiol.1999,87:1313-1316.
24.Heled Y,Moran DS,Mendel L,et al.Human ACE I/D polymorphism is associated with individual differences in exercise heat tolerance.J Appl Physiol.2007,7:72-76.
25.Alvarez R,Terrados N,Ortolanv R,et al.Genetic variation in the rennin-angiotension system and athletic performance.EurJ Appl physiol,2000,82:117-120.
26.Alvarez R,Reguero JR,Batalla A,et al.Angiotensin-converting enzyme and angiotensin Ⅱ recetptor Ⅰ polymorphisms:association with early coronary disease.Cardiovasc Res,1998,40:375-379.
27.Chen Yamin,Offer Amir,Moran Sagiv,et al.ACE ID genotype affects blood creatine kinase response to eccentric exercise.J Appl Physiol,2007,103:2057-2061.
28.Amir O,Amir R,Yamin C,et al.Human,environmental & exercise:the ACE deletion allele is associated with Israeli elite endurance athletes.Exp Physiol,2007,92:881-886.
29.Roman BB,Wieringa B,Koretsky AP.Functional equivalence of creatine kinase isoform in mouse skeletal muscle.J Biol Chem,1997,272:17790-17794.
30.Zhang J,Yu KF.What's the relative risk?A method of correcting the odds ratio in cohort studies of common outcomes.JAMA,1998,280:1690-1691.
31.Saks V,Kaambre T,Guzun R,et al.The creatine kinase phosphotransfer network:thermodynamic and kinetic considerations,the impact of the mitochondrial outer membrane and modelling approaches.Subcell Biochem,2007,46:27-65.
32.Robinson R,Carpenter D,Shaw MA,et al.Mutations in RYRl in malignant hyperthermia and central core disease.Hum Mutat,2006,27:977-989.
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[1]Lin H,Chie W,Lien H.Epidemiological analysis of factors influencing an episode of exertional rhabdomyolysis in high school students[J].Am J Sports Med,2006,34(3):481-486.
[2]Totsuka M,Nakaji S,Suzuki K,et al.Break point of serum creatine kinase release after endurance exercise[J].J Appl Physiol,2002,93(4):1280-1286.
[3]Heled Y,Bloom MS,Wu T J,et al.CK-MM and ACE genotypes and physiological prediction of the creatine kinase response to exercise[J].J Appl Physiol,2007,103(2):504-510.
[4]高炳宏,陈佩杰,董强刚,等.上海地区汉族优秀游泳运动员ACE基因I/D多态性研究[J].中国运动医学杂志,2006,25(5):517-521.
[5]席翼,张秀丽,胡扬,等.中国优秀马拉松运动员ACE基因I/D多态性频率分布特征[J].中国运动医学杂志,2006,25(4):391-394.
[6]Chen Yamin,Offer Amir,Moran Sagiv,et al.ACE ID genotype affects blood creatine kinase response to eccentric exercise[J].J Appl Physiol,2007,103(6):2057-2061.
[7]周多奇,胡扬,刘刚,等.中国北方汉族人群肌型肌酸激酶基因(CKMM)A/G多态研究[J].遗传,2005,27(4):535-538.
[8]Heled Y,Moran DS,Mendel L,et al.Human ACE I/D polymorphism is associated with individual differences in exercise heat tolerance[J].J Appl Physiol.2004,97(1):72-76.
[9]Cerit M,Colakoglu M,Erdogan M,et al.Relationship between ace genotype and short duration aerobic performance development[J].Eur J Appl Physiol,2006,98(5):461-465.
2.Moeller J and McKeag D.Exercise-induced rhabdomyolysis.Sports Med Arthrosc Rev,2004,3:274-279.
3.Bagley WH,Yang H,Shah KH.Rhabdomyolysis.Intern Emerg Med,2007,2:210-8.
4.Hurley JK.Severe rhabdomyolysis in well conditioned athletes.Mil Med,1989,154:244-5.
5.Heled Y,Bloom MS,Wu TJ,et al.CK-MM and ACE genotypes and physiological prediction of the creatine kinase response to exercise.J Appl Physiol.2007,103:504-10.
6.Kauwe JS,Wang J,Mayo K,et al.Alzheimer's disease risk variants show association with cerebrospinal fluid amyloid beta.Neurogenetics,2009,10:13-7.
7.Osawa N,Koya D,Araki S,et al.Combinational effect of genes for the renin-angiotensin system in conferring susceptibility to diabetic nephropathy.J Hum Genet,2007,52:143-51.
8.Zakrzewski-Jakubiak M,de Denus S.Teh renin-angiotensin system-related gene polymorphisms in maximally treated Canadian Caucasian patients with heart failure.Br J Clin Pharmacol.2008,65:742-51.
9.Myerson S,Hemingway H,Budget R,et al.Human angiotensin I converting enzyme gene and endurance performance.J Appl Physiol.1999,87:1313-1316.
10.Heled Y,Moran DS,Mendel L,et al.Human ACE I/D polymorphism is associated with individual differences in exercise heat tolerance.J Appl Physiol.2007,7:72-76.
11.Alvarez R,Terrados N,Ortolanv R,et al.Genetic variation in the rennin-angiotension system and athletic performance.Eur J Appl physiol,2000, 82:117-120.
12.Alvarez R,Reguero JR,Batalla A,et al.Angiotensin-converting enzyme and angiotensin II recetptor I polymorphisms:association with early coronary disease.Cardiovasc Res,1998,40:375-379.
13.Chen Yamin,Offer Amir,Moran Sagiv,et al.ACE ID genotype affects blood creatine kinase response to eccentric exercise.J Appl Physiol,2007,103:2057-2061.
14.Amir O,Amir R,Yamin C,et al.Human,environmental & exercise:the ACE deletion allele is associated with Israeli elite endurance athletes.Exp Physiol,2007,92:881-886.
15.Roman BB,Wieringa B,Koretsky AP.Functional equivalence of creatine kinase isoform in mouse skeletal muscle.J Biol Chem,1997,272:17790-17794.
16.Zhang J,Yu KF.What's the relative risk?A method of correcting the odds ratio in cohort studies of common outcomes.JAMA,1998,280:1690-1691.
17.Saks V,Kaambre T,Guzun R,et al.The creatine kinase phosphotransfer network:thermodynamic and kinetic considerations,the impact of the mitochondrial outer membrane and modelling approaches.Subcell Biochem,2007,46:27-65.
18.Robinson R,Carpenter D,Shaw MA,et al.Mutations in RYR1 in malignant hyperthermia and central core disease.Hum Mutat,2006,27:977-989.
19.Roth SM,Martel GF,Rogers MA.Muscle biopsy and muscle fiber hypercontraction:a brief review.Eur J Appl Physiol,2000,83:239-45.
20.Visweswaran P,Guntupalli J.Rhabdomyolysis.Crit Care Clin,1999,15:415-28.
21.Byrnes WC,Clarkson PM,White JS,et al.Delayed onset muscle soreness following repeated bouts of downhill running.J Appl Physiol.1985,59:710-5.
22.Schwane JA,Johnson SR,Vandenakker CB,et al.Delayed-onset muscular soreness and plasma CPK and LDH activities after downhill running.Med Sci Sports Exerc.1983,15:51-6.
23.Clarkson PM,Nosaka K,Braun B.Muscle function after exercise-induced muscle damage and rapid adaptation.Med Sci Sports Exerc.1992,24: 512-20.
24.Sorichter S,Koller A,Haid C,et al.Light concentric exercise and heavy eccentric muscle loading:effects on CK,MRI and markers of inflammation.Int J Sports Med,1995,16:288-92.
25.Sayers SP,Clarkson PM,Lee J.Activity and immobilization after eccentric exercise:Ⅱ.Serum CK.Med Sci Sports Exerc,2000,32:1593-7.
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