Study On Gene Polymorphism And Expression Of Adenosine Triphosphate Binding Cassette Transporter G2

Part1 Study on the rs2231142(C> A)Polymorphisms of adenosine triphosphate binding cassette transporter G2 in han population of northeastern SichuanObjective: To analyze and compare the distribution frequency of ABCG2 rs2231142(C> A)polymorphism genotypes and alleles in han population of northeastern Sichuan,and to explore the association of this gene polymorphism with gout susceptibility.Methods:(1)169 cases of primary gouty arthritis patients and 193 cases of healthy physical examination were enrolled into the study.The Peripheral blood,clinical data of primary gouty arthritis patients and healthy physical examination were collected.(2)119 cases of gout patients with complete clinical data were used for grouping,which was according to whether or not tophus were divided into 14 cases of gouty tophus group and 105 cases of non-gouty tophus group;according to the incidence of gout sooner or later were divided into 52 cases of early-onset of gout group and 67 cases of late-onset of gout group.(3)To detect the genotypes frequencies of all the research objects by using TaqMan?R probe method Real-time fluorescence quantitative PCR.(4)Statistical analysis was performed using SPSS22.0 statistic analysis software.Chi-square test and calculate the relevant odds ratio(OR)and 95% confidenve interval(95%CI)were used for statistical analysis.P value of <0.05 was considered to indicate a significant difference.Results:(1)Genotypes distribution were accords with the law of Hardy-Weinberg equilibrium in all the research objects(P>0.05).(2)AA genotype frequency in the gout group was significantly higher than the controls group[36.1%,5.7%;P < 0.01;OR=9.345,95%CI(4.712,18.534)].CA genotype frequency in the gout group was significantly higher than the controlsgroup[45.0%,32.6%;P0.05;OR=1.686,95%CI(1.100,2.585)].CC genotype frequency in the gout group was significantly lower than the control group[18.9%,61.7%;P < 0.01;OR=0.145,95%CI(0.090,0.235)].A and C allele in the gout group compared with the control group,there was a statistically significant difference between the two[58.6%,22.0%;41.4%,78.0%;P<0.001;OR=5.008,95%CI(3.623,6.923)].(3)AA genotype frequency in the gouty tophus group was significantly higher than the non-gouty tophus group[71.4%,33.3%;P<0.01;OR=5.000,95%CI(1.464,17.080)].There was no significant difference in CA and CC genotype frequency between the gouty tophus group and non-gouty tophus group(P>0.05,respectively).A and C allele in the gouty tophus group compared with the non-gouty tophus group,there was a statistically significant difference between the two[85.7%,57.1%;14.3%,42.9%;P=0.004;OR=4.500,95%CI(1.508,13.427)].(4)AA genotype frequency in the early-onset of gout group was significantly higher than the late-onset of gout group[51.9%,26.9%;P < 0.01;OR=20.622,95%CI(9.728,43.716)].CA genotype frequency in the early-onset of gout group was significantly lower than the late-onset of gout group[32.7%,53.7%;P < 0.05;OR=0.418,95%CI(0.197,0.888)].There was no significant difference in CC genotype frequency between the early-onset of gout group and late-onset of gout group(P>0.05).A and C allele in the early-onset of gout group compared with the late-onset of gout group,there was a statistically significant difference between the two[66.4%,53.7%;33.6%,46.3%;P=0.047;OR=1.698,95%CI(1.004,2.872)].Conclusion: The polymorphism of ABCG2 rs2231142(C> A)may be related to the incidence of primary gout in han population of northeastern Sichuan.The AA and CA genotype of carrying the A allele is more susceptible to gout,and CC genotype,C allele may have protective effect on the incidence of gout.Subgroup analysis found that AA Genotypes increased the risk of tophus and early-onset gout.C allele may have protective effect on the incidence of tophus,and CA genotype,C allele reduced the risk of early-onset of gout.Part2 Study on the expression of adenosine triphosphate binding cassette transporter G2 m RNA and protein in the peripheral blood mononuclear cells of patients with primary Gouty arthritisObjective: To measure the level of ATP-binding cassette sub-family G member 2(ABCG2)m RNA and protein in peripheral blood monocyte(PBMCs)from patients with gouty arthritis(GA)and healthy controls(HC),and to discuss the role of ABCG2 in the pathogenesis of GA.Methods:(1)82 GA and 46 HC were enrolled into the study.The Peripheral blood,clinical and laboratory data of AG and HC were collected.(2)According to the clinical symptoms of gout patients were divided into 41 cases of acute gouty arthritis(AGA group)and 41 cases of intermittent gouty arthritis(IGA group);According to the level of serum uric acid of above gout patients were divided into 31 cases of UA <420?mol / L gout group(NUA group)and 51 cases of UA> 420?mol / L gout group(HUA group);(3)RT-q PCR and Western blot was employed to study the expression of ABCG2 m RNA and protein in PBMCs.Spearman's correlations were used to evaluated relationship among ABCG2 and the index which were detected.(4)Statistical analysis was performed using SPSS22.0 statistic analysis software.Independent-Samples T test and One-Way ANOVA or Wilcoxon test and Spearman's correlations were used for statistical analysis.P value of <0.05 was considered to indicate a significant difference.Results:(1)The expression levels of ABCG2 m RNA and portein in the PBMCs of IGA patients significantly increased compared to AGA patients(P<0.05,P<0.01)and HC(P<0.01,P<0.05),there was no significant difference in the difference between AGA group and HC group(P>0.05,respectively);(2)The expression levels of ABCG2 m RNA and portein in the PBMCs of HUA patients significantly increased compared to NUA patients(P<0.01,respectively)and HC(P<0.01,P<0.05),there was no significant difference in the difference between NUA group and HC group(P>0.05,respectively);(3)The expression levels of ABCG2 m RNA in the PBMCs was correlated with s UA(r=0.235,P<0.05),negative correlation with creatinine(r=-0.227,P<0.05),no correlation with ESR(r =-0.181,P> 0.05)and high sensitivity C reactive protein(r = 0.027,P> 0.05).(4)There was no significant difference in age composition ratio between GA group and HC group(P> 0.05).The GA group WBC,neutrophil(GR)and mononuclear cells(Mo),TG,TC,LDL,s UA,GLU,creatinine and hs CRP were significantly higher than the normal control group(P < 0.05,respectively),while HDL was significantly lower than the control group(P < 0.01).Conclusion:(1)ABCG2 was expressed in PBMCs and participates in the uric acid transport and was not associated with gout inflammation;(2)ABCG2 and creatinine was negatively correlated,suggesting that the presence of gout in patients with renal dysfunction may be down ABCG2 in PBMCs expression,which may be beneficial to the protection of the kidney,the specific mechanism needs further study.(3)Gout patients was more prone to blood lipids,blood glucose and renal dysfunction compared with the normal population.The changes of blood lipid in gout patients are similar to those of atherosclerosis,it may be accelerated the occurrence of atherosclerosis and cardiovascular and cerebrovascular diseases.