Study On The Distribution Of Body Constitution In Patients With Anti-tuberculosis Drug-induced Liver Injury And Its Correlation With ABC Transporter SNP

BackgroundAnti-tuberculosis drug-induced liver injury(ATDILI)is one of the most common and serious adverse drug reactions in clinical practice,and it is also one of the most important drug-induced liver injury(DILI)in China,with an incidence rate of more than 30%,which can lead to liver failure and even death of patients.It is important to identify risk factors for early detection and prevention of ATDILI.Traditional Chinese medicine(TCM)believes that the physical state determines the susceptibility of the disease,and is of great value to the prognosis of the disease and to guide the diagnosis and prevention.ABC transporter is an important phase III drug transporter,and its genetic polymorphism can lead to changes in pharmacodynamics and pharmacokinetics.Finding rules from the perspective of ATDILI constitution and genetic susceptibility factors,exploring the susceptibility of ATDILI and its possible host genetic mechanism are important for revealing the connotation of TCM constitution,and providing scientific theoretical reference for the use of TCM advantages in clinical diagnosis and treatment and disease prevention significance.Objective①To explore the susceptibility genetic factors of ATDILI with study of the correlation between ABC transporter gene polymorphism(SNP)and ATDILI;②To explore the ATDILI susceptible TCM constitutions with the study of the distribution of ATDILI patients’ TCM constitutions;③To explore the possible host genetic mechanisms’ of the ATDILI susceptible TCM constitutions by analyzing the correlation between ATDILI susceptible TCM constitutions and ATDILI susceptibility genetic factors.MethodsThis study used a case-control study.A total of 643 patients were enrolled.Among them,289 patients who took anti-tuberculosis drugs had liver damage(ATDILI group),and 354 patients who took anti-tuberculosis drugs without liver damage(non-ATDILI)group).Collect and sort out the case information,clinical test indicators and blood samples of the research subjects,extract DNA from the blood samples,and screen the SNP detection sites of ABC transporters;use Sequenom MassARRAY mass spectrometry technology to detect SNPs at 64 sites of ABC transporters in all patients;Select 51 loci that meet Hardy-Weinberg genetic balance for statistical analysis.Chi-square test,multiple logistic regression analysis,Haploview,etc.were used to detect alleles,genotypes,genetic models,LD linkage disequilibrium detection and the association between haplotypes and ATDILI to screen ATDILI susceptible SNP loci.Based on Professor Wang Qi’s 9 TCM constitutions classification criteria,the two groups of people’s physique distribution were investigated and summarized.The Chi-square test,Kruskal Wallis rank sum test,and the multiple Logistic regression analysis method were used to analyze ATDILI’s main physique distribution and concurrent analysis.The physique distribution characteristics were analyzed,and the clinical characteristics of ATDILI susceptible physique were analyzed by layers.Multivariate Logistic regression analysis was used to analyze the association between susceptible physique and susceptible SNP loci of ATDILI;the 2×4 bifurcation analysis method was used to analyze the influence of physique-gene interaction on ATDILI.Results1.The risk of ATDILI in patients with the CC genotype at the rs28656907(T>C)locus of the ABCB1 gene is reduced by 0.58 times(OR=0.58,95%CI:0.36-0.93,P<0.05),both dominant and additive genetic models are the protect the genetic models.Hierarchical analysis showed that the TC and CC genotypes at this locus were the protective factors for abnormal liver biochemical tests of ATDILI(TC genotype:OR=0.57,95%CI:0.35-0.93,P<0.05;CC genotype:OR=0.48,95%CI:0.23-0.97,P<0.05),both additive and dominant genetic models are protective genetic models for abnormal liver damage in liver biochemical examination(additive:OR=0.48,95%CI:0.23-0.99,P<0.05;Dominant:OR=0.54,95%CI:0.34-0.86,P<0.01).Haplotype analysis showed that among the haplotypes composed of rs4148727(A>G)-rs28656907(T>C)-rs10261685(A>C)in the ABCB1 gene,the risk of ATDILI in carriers of the ACA haplotype was reduced(OR=0.77,95%CI:0.56-0.98,P<0.05),while the risk of ATDILI in ATA haplotype carriers increased(OR=1.34,95%CI:1.07-1.83,P<0.01).2.The risk of ATDILI is reduced under the recessive inheritance model of ABCB4 gene rs2071645(G>C)(OR=0.50,95%CI:0.25-0.98,P<0.05),but alleles,genotypes and ATDILI subtypes Hierarchical analysis did not find any correlation with ATDILI,and the results need to be further verified.3.The AG and GG genotypes at rs3829888(A>G)of the ABCB11 gene are ATDILI susceptibility genotypes(AG genotype:OR=1.97,95%CI:1.25-3.12,P<0.01;GG genotype:OR=8.67,95%CI=1.02-73.54,P<0.05),the additive and dominant genetic models are both ATDILI risk genetic models;hierarchical analysis shows that the AG genotype at this locus is hepatocellular injury and cholestasis The risk genotypes of the three subtypes of ATDILI and the mixed type(hepatocellular injury type:OR=2.15,95%CI:1.19-3.91,P<0.05;cholestasis type:OR=2.09,95%CI:1.14-3.83,P<0.05;mixed type:OR=3.77,95%CI:1.92-7.39,P<0.001),GG genotype is the extremely high risk genotype of cholestatic ATDILI,(OR=29.20,95%CI:3.08-276.67,P<0.01);Dominant inheritance models are all three subtypes of ATDILI pathogenesis genetic models(hepatocellular injury type:OR=2.15,95%CI:1.19-3.88,P<0.05;cholestasis type:OR=2.70,95%CI:1.22-5.97,P<0.05;mixed type:OR=3.90,95%CI:2.01-7.56,P<0.001);additive and recessive genetic models are cholestasis type ATDILI with extremely high risk inheritance Model(additive:OR=27.96,95%CI:2.83-276.74,P<0.01;recessive:OR=26.87,95%CI:2.85-253.17,P<0.01).The risk of ATDILI in patients with CT genotype at the rs2216504(C>T)site of the ABCB11 gene increased by 1.67 times(OR=1.67,95%CI:1.02-2.73,P<0.05).The dominant model is the genetic model of ATDILI risk(OR=1.63,95%CI:1.01-2.64,P<0.05),stratified analysis showed that the risk of mixed ATDILI in patients with CT genotypes at this site increased by 2.40 times(OR=2.40,95%CI:1.13-5.09,P<0.05);The dominant inheritance model is a mixed ATDILI risk genetic model(OR=2.38,95%CI:1.15-4.92,P=0.019).Haplotype analysis showed:ABCB11 gene rs10930343(AAA>DEL)-rs3829888(A>G)-rs3814382(G>A)-rs3814381(C>T)-rs2216504(C>T)-rs4148765(C>T)The risk of AAAGGCTC haplotype ATDILI was increased by 1.71-fold(OR=1.71,95%CI:1.09-3.25,P=0.025)in haploids composed of dots.4.The risk of ATDILI in carriers of the GA genotype at the rs4148211(G>A)locus of the ABCG8 gene increased by 1.70 times(OR=1.70,95%CI:1.14-2.53,P<0.01);the dominant inheritance model is its risk inheritance model(OR=1.55,95%CI:1.05-2.27,P=0.026).Layer analysis showed that the risk of cholestatic DILI in patients with GA genotype at this site increased by 3.15 times(OR=3.15,95%CI:1.56-6.35,P=0.001),and the risk of mixed DILI increased by 2.87 times(OR=2.87),95%CI:1.52-5.41,P<0.01);the dominant genetic model is a risk factor for the onset of cholestasis and mixed ATDILI(cholestasis:OR=2.75,95%CI:1.38-5.51,P<0.01;Mixed type:OR=2.52,95%CI:1.35-4.70,P<0.01).5.There was no correlation between ABCC2,ABCC4,ABCC10 and ABCG2 gene polymorphisms and ATDILI(P>0.05).6.Analysis of the main physique distribution of ATDILI patients shows that:The main physique distribution of ATDILI patients is mainly qi deficiency(25.95%),damp heat(20.76%),yin deficiency(18.69%)and phlegm dampness(14.88%);Compared with the ATDILI group,the ATDILI group showed a significant decrease in the proportion of peace and quality,while the distribution ratio of Qi deficiency and damp-heat quality increased significantly,both with significant statistical differences(P<0.01);multiple logistic regression analysis showed that the risk of ATDILI in patients with damp-heat quality increased by 1.93 times(OR=1.93,95%CI:1.25-2.97),the risk of ATDILI in patients with Qi deficiency and constitution increased 2.1 times(OR=2.10,95%CI:1.40-3.15),and moderate quality is a strong protective factor for the risk of ATDILI(OR=0.21,95%CI:0.10-0.41),all have significant statistical significance(P<0.01).7.The distribution analysis of the constitution of ATDILI patients with clamp showed that 73.36%of ATDILI patients had a biased physique with clamp,which was significantly higher than that of non-ATDILI patients(P<0.01).Qi deficiency,damp-heat,yin-deficiency,and phlegm-dampness accounted for ATDILI patients’ 80.27%.The proportions of the four high-frequency physiques combined with stagnation of Qi were significantly increased in ATDILI patients(P<0.05),accounting for 21.12%(49:232)of these four high-frequency constitutions,which is ATDILI’s easy Sense and physique.8.The analysis of the clinical characteristics of ATDILI patients’ susceptible physique showed that the distribution of Qi-deficiency patients in ATDILI patients was positively correlated with age and incubation period.The blood TP level was lower than that of patients with combined qi stagnation,and the TBA level was higher than that of damp-heat patients.The differences were statistically significant(P<0.05);patients with damp-heat quality were mainly distributed in people under 30 years of age or people with moderate to severe liver injury(P<0.05),and their distribution ratio was negatively correlated with the course of the disease(P<0.05)The blood ESR and CRP levels of patients with damp-heat type were significantly higher than those of patients with combined qi stagnation(P<0.05);the distribution of moderate to severe liver injury in patients with combined qi stagnation was significantly increased(P<0.01),and their blood TBA The level was significantly higher than that of patients with damp-heat,and the ALT level was significantly higher than that of patients with damp-heat and Qi-deficiency.The difference was statistically significant(P<0.05).9.Carriers of the ABCB11_rs3829888 locus additive genetic model have a 2.29-fold increase in risk of Qi-deficiency ATDILI(OR=2.29,95%CI:1.23-4.28,P<0.01),and carriers of the ABCB11_rs4148211 locus additive genetic model have Qi-deficiency ATDILI The risk increased by 2.37 times(OR=2.37,95%CI:1.34-4.18 P<0.01);carriers of the ABCB11_rs2216504(C>T)locus additive genetic model had a 2-fold increase in risk of ATDILI(OR=2.00,95%)CI:1.03-3.89,P<0.05).In addition,no correlation was found between the calm quality and the combined Qi stagnation quality and the SNP locus of the ABC transporter(P>0.05).10.There is a negative interaction between the ABCB1_rs28656907 locus and the ABCB1_rs28656907 site.Based on the additive model,there is a negative interaction(RERI=0.55,S<1,P<0.01).The risk of ATDILI in patients with the ABCB1_rs28656907 locus additive genetic model is reduced by 0.23 times.(95%CI:0.100-0.536);In addition,there was no interaction between Qi deficiency,damp-heat and combined Qi stagnation and ABC transporter susceptible SNP sites(P>0.05).Conclusions1.The SNP at the rs28656907(T>C)locus of the ABCB1 gene may be a susceptibility protection factor for ATDILI;the rs2071645(G>C)locus of the ABCB4 gene only reduces the risk of ATDILI under the recessive genetic model,and the results need to be verified;2.SNPs at ABCB11 gene rs3829888(A>G),rs2216504(C>T)locus and ABCG8 gene rs4148211(G>A)locus may be ATDILI susceptibility risk factors.3.No association between ABCC2,ABCC4,ABCC10 and ABCG2 gene polymorphisms and ATDILI was found.4.The main physiques of the ATDILI group are mainly qi deficiency,damp-heat,yin deficiency and phlegm dampness.Among them,the qi deficiency,damp-heat and qi stagnation are ATDILI susceptible and dangerous constitutions,and the peaceful constitution is ATDILI susceptible and protective constitution.5.The clinical characteristics of patients in the ATDILI group who are susceptible to physique:Most patients with Qi deficiency are middle-aged and elderly patients,with relatively slow onset,low blood TP levels and high TBA levels;most patients with damp-heat type are young,with relatively short course of disease and moderate to severe liver disease The proportion of blood damage is relatively high,and the blood ESR and CRP levels are relatively high;the proportion of moderate to severe liver damage is relatively high in patients with qi stagnation,and the blood TBA and ALT levels are relatively high.6.SNPs at the ABCB11_rs3829888(A>G)locus and ABCG8_rs4148211(G>A)may be risk factors for ATDILI patients with Qi deficiency;ABCB11_rs2216504(C>T)may be risk factors for ATDILI patients with damp-heat type of constitution.7.There is a positive interaction between the peaceful constitution and SNP at ABCB1_rs28656907 locus,the risk of ATDILI is reduced in patients with the peaceful constitution and the additive genetic model at ABCB1_rs28656907 locus.