Association Of Synergistic Effects Of Wuzhi Capsules On Tacrolimus And CYP3A4,CYP3A5,MDR1 And PXR Polymorphisms

Background and ObjectivesTacrolimus has a strong immunosuppressive effect and few adverse reactions.It is widely used as first-line immunosuppressive agent to prevent rejection after organ transplantation.Our previous studies found that wuzhi capsules can significantly improve the plasma concentration of tacrolimus for the first time.A large number of studies have confirmed that the CYP3A5a*3 genotype is strongly associated with tacrolimus plasma levels,which is the most consistent conclusion in the study of tacrolimus.For the study of the correlation between CYP*18B,MDR1 PXR gene polymorphism and tacrolimus blood concentration,the current research opinions are not uniform.There are a lot of in vitro experiments to confirm that wuzhi capsules improve tacrolimus blood concentration by inhibiting the CYP3A and P-gp,However,whether the interaction between wuzhi capsule and tacrolimus is related to polymorphisms of CYP3A,MDR1,PXR and so on,has not yet been reported.A retrospective analyse was used to study the correlation of CYP3A4,CYP3A5,MDR1 and PXR gene polymorphisms with the effect of wuzhi capsules on tacrolimus in the Chinese renal transplant recipients..Research MethodsA total of 74 patients were continually administered with wuzhi capsules as WZC(+)group.A total of 96 patients took tacrolimus alone as WZC(-)group.CYP3A4*18B,CYP3A5*3,MDR1 1236C>T,MDRI 2677G>T/A,MDR1 3435C>T polymorphisms were determined by restriction fragment length polymorphism(RFLP)analysis.The PXR 7635A>G and PXR 24381A>C genotypes were determined by PCR-direct sequencing.PXR 6bp deletions genotypes was determined by Allelic Special-Touch down PCR.The FK506 whole blood levels of renal transplant recipients were measured by chemiluminescent microparticle immunoassay.The Kolmogorov-Smirnov test was used to assess distribution.The values were expressed as mean±standard deviation(SD)if they conformed to a normal distribution;otherwise,median(quartiles 1-3)was applied.Statistically significant difference between two populations were assessed using the independent samples t-test if they showed homogeneity of variance;otherwise,the Mann-Whitney U test was applied.The chi-square test or Fisher ’ s exact tests was used to examine differences in count data.Analysis of covariance(ANCOVA)was performed after In-transformed tacrolimus C0/D(InC0/D)which showed homogeneity of variance and conformed to a normal distribution,with RBC and tacrolimus C0/D on 0 month as independent covariates.Spearman’s correlation was used to analyze the correlation between various factors and tacrolimus C0/D.ResultsFor CYP3A5*3 genotype,on 1 month,tacrolimus Ca/D of patients with GG genotype in wuzhi capsule(+)group was significantly higher than that in wuzhi capsule(-)group of the same genotype(P=0.022),but on 3 months to 3 years,the differences were no statistical significance(P>0.05).On 1 months to 3 years,tacrolimus C0/D of patients with GA/AA genotype in wuzhi capsule(+)group was significantly higher than that of the patients with the same genotype in wuzhi capsule(-)group(P<0.001).For PXR7635 G>A genotype,tacrolimus C0/D of patients with GG was significantly higher than those with GA/AA in wuzhi(+)group on 3 months to 2 years and there was no significant difference in wuzhi capsule(-)group,whether Co-administeration of wuzhi capsules and tacrolimus or not,tacrolimus C0/D of CYP3A4*18B,MDR1 1236C>T,MDR1 2677G>T/A,MDR1 3435C>T,PXR 6bp deletions,PXR 24381 A>C genotype was not significantly different.The correlation between the factors and the concentration of tacrolimus C0/D was analyzed with Sperman’s correlation.In wuzhi capsule(-)group,The factors of Sex,ALT,RBC,Hb,CYP3A4*18B,CYP3A5a*3 and MDR1 2677G>T/A related to tacrolimus C0/D.In wuzhi(+)group,only the factor of PXR7635G>A related to tacrolimus C0/D.Conclusions1.Wuzhi capsule can significantly improve tacrolimus C0/D in CYP3A5*3 GA/AA genotype,but no significant effect on patients with CYP3A5a*3GG genotype.2.When combined with wuzhi capsules,PXR 7635 G>A mutation was associated with tacrolimus C0/D.3.When combined with wuzhi capsules,CYP3A4*18B,MDR1 1236C>T,MDR1 2677G>T/A,MDR1 3435C>T,PXR 6bp deletions,PXR 24381A>C polymorphisms were not associated with tacrolimus C0/D.