| ObjectiveType 2 diabetes mellitus(T2DM)combined with lower extremity arterial disease(LEAD)is a common complication of diabetes mellitus.The risk of LEAD in patients with type 2 diabetes was significantly higher than that in non-diabetic patients.The onset of the disease is early,the severity of the disease is severe,and the treatment is difficult.LEAD is a complex disease caused by environmental and genetic factors.Gene polymorphism,many non-genetic factors(such as age,duration of diabetes,smoking,dyslipidemia,hyperglycemia,and hypertension)play an important role in the development of LEAD,but the results vary.At present,most studies have shown that leptin(LEP)and leptin receptor(LEPR)gene polymorphism are closely related to obesity,hypertension,cardiovascular and cerebrovascular diseases.Now,there is no report on the effects of LEP and LEPR gene polymorphism and the interaction with non-genetic factors on lower extremity arterial diseases.Objects and methods1.Selection of research objectsA total of 789 patients with type 2 diabetes who were admitted to the Department of endocrinology of the Affiliated Hospital of North China University of Science and technology from May 2015 to November 2016 were selected as subjects,380 cases of LEAD,and 409 cases of non-LEAD.2.Laboratory testThe fasting venous blood were collected for 8ml to detect the levels of the total cholesterol(TC),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),triglyceride(TG),Hb Alc and leptin(LEP).3ml venous blood in EDTA anticoagulant tube was used for DNA extraction and polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)to detect LEP gene G2548 A locus,rs3828942 locus,LEPR gene Gln223 Arg locus and Prol019 Pro locus polymorphism.3.Statistical analysisEpidata 3.0 was used to establish the data base of the patients with diabetesmellitus,and the data were analyzed with SPSS 17.0.The quantitative data are expressed as(x ± s).The t test was used for comparison between two groups,and the qualitative data were analyzed by ? 2 test.The risk factors were analyzed by multivariate logistic regression analysis.The effects of environment and gene interaction of LEP and LEPR gene on LEAD were analyzed by cross analysis.Experimental results1.Relationship between non-genetic risk factors and LEADAfter the comparison of the basic data of LEAD patients with that of non-LEAD patients,Hb A1c,TC,LDL-C,Leptin,diabetes,hypertension,coronary heart disease and cerebral infarction were statistically significant(P<0.05).There was no significant difference in age,sex,BMI,TG,HDL,and smoking in patients with LEAD and non-LEAD(P>0.05).The prevalence of LEAD increased with age in patients with diabetes mellitus,and after the chi square test it was statistically significant(?2??=108.874,P<0.001).Multivariate logistic regression showed that:Age,high TC blood syndrome,high LDL-C blood syndrome,high Hb Alc levels,duration of diabetes,hypertension,coronary heart disease,cerebral infarction,and the increase of leptinis were associated independently with risk of LEAD(P<0.005).2.Relationship between leptin and leptin receptor gene polymorphism and LEADThe distributions of LEP gene 2548G/A locus AA,AG,GG genotype in non-LEAD group were: 44.3%,42.3%,13.4%,and those in LEAD group were:34.7%,44.7%,20.5%.There was a statistical difference between the two groups(?2=10.623,P =0.005).The G allele frequency(38.6%)in the LEAD group was higher than that in the control group(34.6%),and the difference was statistically significant(?2=11.447,P=0.001).Multivariate regression analysis showed that the risk of LEAD in patients carrying LEP gene 2548G/A locus GG genotype was 1.998(95%CI:1.316~3.017)times of those carrying AA genotype.GG genotype may be a susceptible genotype of LEAD.The distributions of LEP gene rs3828942 locus AA,AG,GG genotype in non-LEAD group were: 45.5%,43.0%,11.5%,and those in LEAD group were:43.2%,43.7%,13.2%.There was no statistical difference between the two groups(?2=0.703,P=0.704).The G allele frequency(35.0%)in the case group was higher than that in the control group(33.0%),and the difference was not statistically significant(?2=0.697,P=0.404).Multivariate analysis showed that rs3828942 polymorphism was not associated with LEAD.The distributions of LEP gene Gln223 Arg locus AA,AG,GG genotype in non-LEAD group were: 10.3%,39.4%,50.4%,and those in LEAD group were: 6.3%,35.3%,58.4%.There was a statistical difference between the two groups(?2=6.922,P=0.031).The G allele frequency(76.1%)in the LEAD group was higher than that in the control group(70.0%),and the difference was statistically significant(?2=7.195,P=0.007).Multivariate regression analysis showed that the risk of LEAD in patients carrying LEP gene Gln223 Arg locus GG genotype was 1.816(95%CI: 1.100~3.108)times of those carrying AA genotype,suggesting that GG genotype may be a susceptible genotype of LEAD.The distributions of LEP gene Pro1019 Pro locus GG,AG,AA genotype in non-LEAD group were: 16.6%,44.3%,39.1%,and those in LEAD group were:11.3%,41.6%,47.1%.There was a statistical difference between the two groups(?2=7.200,P=0.027).Compared with the control group(61.2%),the A allele frequency(67.9%)in the LEAD group was statistically significant(?2=7.599,P=0.006).Multivariate regression analysis showed that the risk of LEAD in patients carrying LEPR gene Pro1019 Pro locus AA genotype was 1.717(95%CI: 1.109~2.658)times of those carrying GG genotype,suggesting that AA genotype may be a susceptible genotype of LEAD.3.Analysis of the effect of gene-gene interaction on LEADThere was no interaction based on additive models of the 4 polymorphism(P>0.05).4.Analysis of the effect of gene-environment interaction on LEADBy cross analysis,there was an interaction based on additive models between the Gln223 Arg locus and LDL-C(U=2.658,P=0.008);There was positive interaction between Gln223 Arg locus and LDL-C,the synergy index(S)= 5.310,attributable proportion of interaetion(AP)=0.570,AP *=0.812,relative excess risk of interaction(RERI)= 1.918,odds ratio of interaction(ORint)= 2.269.There was an interaction based on additive models between the Gln223 Arg locus and Hb A1c(U=2.152,P=0.031);There was positive interaction between Gln223 Arg locus and Hb A1 c,the synergy index(S)= 7.027,attributable proportion of interaetion(AP)=0.522,AP*=0.858,relative excess risk of interaction(RERI)= 1.332,odds ratio of interaction(ORint)= 2.074.In this study,the 4 polymorphism did not show theinteraction based on additive model with TC,hypertension,coronary heart disease,andcerebral infarction.Conclusions1.High TC,high LDL_C,high Hb Alc,duration of diabetes,age,high leptin,hypertension,coronary heart disease and cerebral infarction are the risk factors of LEAD in diabetic population.2.LEP gene locus G2548 A,LEPR gene locus Gln223 Arg and Prol019 Pro polymorphism are independent risk factors for LEAD in patients with diabetes mellitus.3.LEP gene 2548G/A locus,rs3828942 locus,and LEPR gene Gln223 Arg locus,Pro1019 Pro locus showed no interaction based on additive model.4.There are positive interactions to the patients of LEAD between LEPR gene Gln223 Arg and HbA1 c,LDL-C. |
PDF Full Text Request