The Effect Of Vitamin D Receptor Gene Polymorphism On Amlodipine Blood Concentration And Antihypertensive Response

Objective: To evaluate the effect of VDR FokI gene polymorphisms on concentration and antihypertensive effect of amlodipine.Methods:1. Culture the LS174T cell and HepG2cell, and give different concentrations of1,25-dihydroxyvitamin D3(0.01μmol/L、0.05μmol/L0.25μmol/L) to interfere, which is a vitamin D receptor agonist. After incubating for24、36、48hours, Using real-time fluorescence quantitative PCR to determin the mRNA level of VDR and CYP3A4and observe the effect of VDR agonists on CYP3A4mRNA expression.2.112patients with mild or moderate primary hypertension were selected randomly, after treatment of4weeks with amlodipine, there are60patients completing the trial. Measuring blood pressure before treatment and after4weeks administration, analysising genotyping of VDR FokI (T>C) and CYP3A4*1G (G> A) through using the polymerase chain reaction (PCR) product direct sequencing method; HPLC-MS/MS method determinate serum concentrations of amlodipine, evaluating the effect of VDR, FokI(T> C) and CYP3A4*1G (G> A) gene polymorphisms on amlodipine concentration and antihypertensive effect.Results:1. The impact of VDR agonists on expression level of CYP3A4in LS174T/HepG2cells:In LS174T cells, drug intervention group CYP3A4expression levels were significantly higher compared with the control group(P<0.05);drug upregulate the expression level of CYP3A4in the0-0.05μmol/L concentration with a dose-dependent manner; drug upregulate the expression level of CYP3A4within the range of0h-36h with a time-dependent correlation; in HepG2cells, CYP3A4expression level of the drug intervention group was significantly higher than the control group (P<0.05), but have no time and dose dependent manner.2.VDR FokI(T>C) gene polymorphism of recipients with hypertensive population and the effect on plasma concentration and antihypertensive effects of amlodipine:Among60patients with hypertension, the frequency of VDR FokI(T>C) genotypes of wild-type (TT) is the lowest (26.7%), followed by the heterozygous (TC)(30%), and mutant homozygous (CC) have the highest frequency(43.3%). The C allele frequency (58.3%) is higher than the T allele frequency (41.6%). As for the VDR FokI sites of TT, TC and CC genotype, plasma concentration showed no trend, three group have no significant difference in different time points (P>0.05). The bit point mutation of VDR FokI have no significant correlation with the antihypertensive effect of amlodipine.4. Correlation analysis of the effect VDR FokI and CYP3A4*1G gene polymorphism on amlodipine blood drug concentration and antihypertensive effect:In the patients with CYP3A4GA/AA, blood drug concentration of VDR FokI TT, TC, CC type have a trend of decreasing, but no significant difference. TT, TC, CC in patients on antihypertensive efficacy rates were100.00%、37.50%and71.43%respectively. There was significant difference between group comparison (P<0.05).Conclusions:1. VDR agonists can upregulate CYP3A4expression in LS174T cells and HepG2cells;2. Gene polymorphism of VDR FokI have no significant impact on plasma concentration and antihypertensive effects of amlodipine.