线粒体基因A3243G突变阳性糖尿病患者的临床特点分析
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摘要
目的探讨线粒体基因(mtDNA)A3243G突变阳性糖尿病患者的血糖、血压特征,及其与mtDNA突变负荷间的关系。方法选择福建医科大学附属第一医院2006-2017年初筛的mtDNA A3243G突变阳性的20例糖尿病患者及20例mtDNA A3243G突变阴性的糖尿病患者,通过直接测序进一步确认致病性突变。收集这些糖尿病患者的血糖、血压等临床资料并进行分析总结。对突变阳性患者外周血和(或)尿沉渣组织的DNA进行聚合酶链反应(PCR)-限制性片段长度多态性(RFLP)分析,计算突变负荷(即突变型mtDNA所占的比例)。结果经测序确认为mtDNA A3243G突变阳性的糖尿病患者20例,其中9例为单纯糖尿病,11例合并线粒体脑肌病。mtDNA A3243G突变阳性组的体质量指数(BMI)、糖化血红蛋白低于对照组[(17.81±2.07)比(25.23±4.01)kg/m~2,(7.56±2.18)%比(9.58±2.50)%;均P<0.01],血乳酸高于对照组[(3.60±1.97)比(1.72±0.40)mmol/L,P<0.05]。单纯糖尿病患者的突变负荷、舒张压、血乳酸水平低于糖尿病合并脑肌病患者[(15.03±6.03)%比(55.84±14.67)%,(69.6±9.7)比(78.0±7.5)mm Hg,(1.94±0.25)比(4.49±1.91)mmol/L,均P<0.05],年龄、BMI高于糖尿病合并脑肌病组[(51.2±15.6)比(32.6±10.0)岁,(19.0±2.1)比(16.8±1.5)kg/m~2,均P<0.05]。同时对6例患者的尿液和血液进行mtDNA A3243G突变定量检测,发现尿液的突变负荷大于血液中的突变负荷。结论 mtDNA A3243G突变单纯糖尿病患者突变负荷低于糖尿病合并脑肌病/肌病患者。对于BMI<20kg/m~2、血乳酸偏高的母系遗传糖尿病患者,很有必要进行mtDNA突变检测,尿沉渣组织更适合可用于mtDNA诊断。
Objective To examine the characteristics of blood glucose and blood pressure in diabetic patients with positive mitochondrial DNA A3243 G mutation and their relationships with mitochondrial DNA mutation load.Methods A total of 40 diabetic patients with(n=20)and without(n=20,control)positive mitochondrial DNA A3243 Gmutation were enrolled from the First Affiliated Hospital of Fujian Medical University between 2006 and2017. Direct sequencing was used to confirm the pathogenic mutation of mitochondrial DNA A3243 G. The clinical data were collected and analyzed. The polymerase chain reaction(PCR)-restriction enzyme digestion analysis was used for quantitative calculation of mutation load which was defined as the proportion of the mutant mtDNA in blood and/or urinary sediment tissue DNA. Results All 20 diabetic patients with positive mtDNA A3243 Gmutation were verified by sequencing. Among these 20 diabetic patients,9 were simple diabetes and 11 complicated with mitochondrial encephalomyopathy. The body mass index(BMI)and haemoglobin A1 c(HbA1 c)in mtDNA A3243 G mutation positive group were lower,while blood lactate level was higher than those in control group[(17.8±2.1)vs(25.2±4.0)kg/m~2,(7.56±2.18)% vs(9.58±2.50)%,and(3.60±1.97)vs(1.72±0.40)mmol/L respectively,all P<0.05]. The mutation load,diastolic blood pressure,and blood lactate levels were lower,while mean age and BMI were higher in simple diabetics than those in diabetics combined with encephalomyopathy [(15.03±6.03)% vs(55.84±14.67)%,(69.6±9.7)vs(78.0±7.5)mm Hg,(1.94±0.25)vs(4.49±1.91)mmol/L,(51.2±15.6)vs(32.6±10.0)years,(19.0±2.1)vs(16.8±1.5)kg/m~2 respectively,all P<0.05]. Quantitative detection of mtDNA A3243 G mutations in urine and blood DNA from 6 patients revealed that in urine mtDNA A3243 G mutations load was greater than in blood. Conclusion In diabetic patients with positive mtDNA A3243 G mutantion,the mutation load in simple diabetics is lower than in diabetics combined with encephalomyopathy. It is necessary to detect mtDNA A3243 Gmutation in maternally inherited diabetic patients with high blood lactate and BMI<20 kg/m~2,and urinary sediment tissue is more suitable for detection of mitochondrial DNA mutation.
Objective To examine the characteristics of blood glucose and blood pressure in diabetic patients with positive mitochondrial DNA A3243 G mutation and their relationships with mitochondrial DNA mutation load.Methods A total of 40 diabetic patients with(n=20)and without(n=20,control)positive mitochondrial DNA A3243 Gmutation were enrolled from the First Affiliated Hospital of Fujian Medical University between 2006 and2017. Direct sequencing was used to confirm the pathogenic mutation of mitochondrial DNA A3243 G. The clinical data were collected and analyzed. The polymerase chain reaction(PCR)-restriction enzyme digestion analysis was used for quantitative calculation of mutation load which was defined as the proportion of the mutant mtDNA in blood and/or urinary sediment tissue DNA. Results All 20 diabetic patients with positive mtDNA A3243 Gmutation were verified by sequencing. Among these 20 diabetic patients,9 were simple diabetes and 11 complicated with mitochondrial encephalomyopathy. The body mass index(BMI)and haemoglobin A1 c(HbA1 c)in mtDNA A3243 G mutation positive group were lower,while blood lactate level was higher than those in control group[(17.8±2.1)vs(25.2±4.0)kg/m~2,(7.56±2.18)% vs(9.58±2.50)%,and(3.60±1.97)vs(1.72±0.40)mmol/L respectively,all P<0.05]. The mutation load,diastolic blood pressure,and blood lactate levels were lower,while mean age and BMI were higher in simple diabetics than those in diabetics combined with encephalomyopathy [(15.03±6.03)% vs(55.84±14.67)%,(69.6±9.7)vs(78.0±7.5)mm Hg,(1.94±0.25)vs(4.49±1.91)mmol/L,(51.2±15.6)vs(32.6±10.0)years,(19.0±2.1)vs(16.8±1.5)kg/m~2 respectively,all P<0.05]. Quantitative detection of mtDNA A3243 G mutations in urine and blood DNA from 6 patients revealed that in urine mtDNA A3243 G mutations load was greater than in blood. Conclusion In diabetic patients with positive mtDNA A3243 G mutantion,the mutation load in simple diabetics is lower than in diabetics combined with encephalomyopathy. It is necessary to detect mtDNA A3243 Gmutation in maternally inherited diabetic patients with high blood lactate and BMI<20 kg/m~2,and urinary sediment tissue is more suitable for detection of mitochondrial DNA mutation.
引文
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[12]Gerbitz KD,van den Ouweland JM,Maassen JA,et al.Mitochondrial diabetes mellitus:a review[J].Biochim Biophys Acta,1995,1271(1):253-260.
[13]Patel MN.Oxidative stress,mitochondrial dysfunction,and epilepsy[J].Free Radic Res,2002,36(11):1139-1146.
[14]Carroll RW,Murphy R.Monogenic diabetes:a diagnostic algorithm for clinicians[J].Genes(Basel),2013,4(4):522-535.
[15]Maassen JA,Janssen GM,Hart LM.Molecular mechanisms of mitochondrial diabetes(MIDD)[J].Ann Med,2005,37(3):213-221.
[16]Strauss DS,Freund KB.Diagnosis of maternally inherited diabetes and deafness(mitochondrial A3243G mutation)based on funduscopic appearance in an asymptomatic patient[J].Br J Ophthalmol,2012,96(4):604.
[17]Wang S,Li R,Fettermann A,et al.Maternally inherited essential hypertension is associated with the novel 4263A>G mutation in the mitochondrial tRNAIle gene in a large Han Chinese family[J].Circ Res,2011,108(7):862-870.
[18]Fayssoil A,Laforêt P,Bougouin W,et al.Prediction of long term prognosis by heteroplasmy levels of the m.3243A>G mutation in patients with the mitochondrial encephalomyopathy,lactic acidosis and stroke-like episodes syndrome[J].Eur J Neurol,2017,24(2):255-261.
[19]Shanske S,Pancrudo J,Kaufmann P,et al.Varying loads of the mitochondrial DNA A3243Gmutation in different tissues:implications for diagnosis[J].Am J Med Genet,2010,130A(2):134-137.
[20]Xia CY,Liu Y,Liu H,et al.Clinical and molecular characteristics in 100Chinese pediatric patients with m.3243A>G mutation in mitochondrial DNA[J].Chin Med J,2016,129(16):1945-1949.
[21]Frederiksen AL,Andersen PH,Kyvik KO,et al.Tissue specific distribution of the 3243A>G mtDNA mutation[J].J Med Genet,2006,43(8):671-677.
[22]林晓贞,陈万金,王柠,等.应用RT ARMS-qPCR技术进行MELAS综合征线粒体DNA A3243G突变负荷研究[J].中华神经科杂志,2009,42(3):197-200.
[23] McDonnell MT,Schaefer AM,Blakely EL,et al.Noninvasive diagnosis of the 3243A> G mitochondrial DNA mutation using urinary epithelial cells[J].Eur J Hum Genet,2004,12(9):778-781.
[2]项坤三.特殊类型糖尿病[M].上海:上海科学技术出版社,2011.
[3]Hannah-Shmouni F,Sirrs S,Mezei MM,et al.Increased prevalence of hypertension in young adults with high heteroplasmy levels of the MELAS m.3243A>G mutation[J].JIMD Rep,2014,12:17-23.
[4]Laloi-Michelin M,Meas T,Ambonville C,et al.The clinical variability of maternally inherited diabetes and deafness is associated with the degree of heteroplasmy in blood leukocytes[J].J Clin Endocrinol Metab,2009,94(8):3025-3030.
[5]de Laat P,Koene S,van den Heuvel LP,et al.Clinical features and heteroplasmy in blood,urine and saliva in 34Dutch families carrying the m.3243A>G mutation[J].J Inherit Metab Dis,2012,35(6):1059-1069.
[6]殷峻,包玉倩.线粒体糖尿病的临床特征与应对[J].中华糖尿病杂志,2017,9(6):342-345.
[7]Bernier FP,Boneh A,Dennett X,et al.Diagnostic criteria for respiratory chain disorders in adults and children[J].Neurology,2002,59(9):1406-1411.
[8]中国血压测量工作组.中国血压测量指南[J].中华高血压杂志,2011,19(12):1101-1115.
[9]Goto Y,Horai S,Mmsuoka T,et al.Mitochondrial myopathy,encephalopathy,lacticacidosis, andstroke-likeepisodes(MELAS):acorrelative study of the clinica1features and mitochondrial DNA mutation[J].Neurology,1992,42(3Pt 1):545-550.
[10]Maassen JA,Essen E,van den Ouweland JMW,et al.Molecular and clinical aspects of mitochondrial diabetes mellitus[J].Exp Clin Endocrinol Diabetes,2011,109(3):127-134.
[11]Nesbitt V,Pitceathly RDS,Turnbull DM,et al.The UK MRC mitochondrial disease patient cohort study:clinical phenotypes associated with the m.3243A>G mutation implications for diagnosis and management[J].J Neurol Neurosurg Psychiatry,2013,84(8):936-938.
[12]Gerbitz KD,van den Ouweland JM,Maassen JA,et al.Mitochondrial diabetes mellitus:a review[J].Biochim Biophys Acta,1995,1271(1):253-260.
[13]Patel MN.Oxidative stress,mitochondrial dysfunction,and epilepsy[J].Free Radic Res,2002,36(11):1139-1146.
[14]Carroll RW,Murphy R.Monogenic diabetes:a diagnostic algorithm for clinicians[J].Genes(Basel),2013,4(4):522-535.
[15]Maassen JA,Janssen GM,Hart LM.Molecular mechanisms of mitochondrial diabetes(MIDD)[J].Ann Med,2005,37(3):213-221.
[16]Strauss DS,Freund KB.Diagnosis of maternally inherited diabetes and deafness(mitochondrial A3243G mutation)based on funduscopic appearance in an asymptomatic patient[J].Br J Ophthalmol,2012,96(4):604.
[17]Wang S,Li R,Fettermann A,et al.Maternally inherited essential hypertension is associated with the novel 4263A>G mutation in the mitochondrial tRNAIle gene in a large Han Chinese family[J].Circ Res,2011,108(7):862-870.
[18]Fayssoil A,Laforêt P,Bougouin W,et al.Prediction of long term prognosis by heteroplasmy levels of the m.3243A>G mutation in patients with the mitochondrial encephalomyopathy,lactic acidosis and stroke-like episodes syndrome[J].Eur J Neurol,2017,24(2):255-261.
[19]Shanske S,Pancrudo J,Kaufmann P,et al.Varying loads of the mitochondrial DNA A3243Gmutation in different tissues:implications for diagnosis[J].Am J Med Genet,2010,130A(2):134-137.
[20]Xia CY,Liu Y,Liu H,et al.Clinical and molecular characteristics in 100Chinese pediatric patients with m.3243A>G mutation in mitochondrial DNA[J].Chin Med J,2016,129(16):1945-1949.
[21]Frederiksen AL,Andersen PH,Kyvik KO,et al.Tissue specific distribution of the 3243A>G mtDNA mutation[J].J Med Genet,2006,43(8):671-677.
[22]林晓贞,陈万金,王柠,等.应用RT ARMS-qPCR技术进行MELAS综合征线粒体DNA A3243G突变负荷研究[J].中华神经科杂志,2009,42(3):197-200.
[23] McDonnell MT,Schaefer AM,Blakely EL,et al.Noninvasive diagnosis of the 3243A> G mitochondrial DNA mutation using urinary epithelial cells[J].Eur J Hum Genet,2004,12(9):778-781.