| [Objective](1)To analysis the distribution of gene polymorphism forCYP3A4*18A、CYP3A4*18B and MDR1C3435T in hyperlipidemia population.(2)To determine the patients`steady plasma concentration of atorvas-tatin by HPLC.(3)To inspect the variations of concentration for totalcholesterol,triglycerides,low-density and high-density lipoproteincholesterol in serum before and after treatment with atorvastatin.(4)Toinvestigate the correlations between gene polymorphisms of CYP3A4*18A、CYP3A4*18B、MDR1C3435T and plasma concentration of atorvastatin or it`sefficacy.The investigation could provide the basic proof for clinicalindividual drug therapy based on genetic background.[METHODS] Refer to2007" Chinese adult dyslipidemia prevention guide",115patients with hyperlipidemia were selected,their while liver andkidney function were normal. Atorvastatin was used as the only lipid-adjusting drugs, therapeutic regimen was20mg qd..1.To acquire the wholeblood from the patients being selected, extract DNA by absorption column,UV method was used to determine the content and purity of DNA;To establishPCR-RFLP method for detection of genotype of CYP3A4*18A, CYP3A4*18Band MDR1C3435T,the PCR products of CYP3A4*18B were sequenced directly toconfirm the reliability of the PCR-RFLP method.The genotypes of selectedpopulation is tested in Hardy-Weinberg genetic equilibrium by chi-squaretest.2.To collect blood samples with steady trough concentration, the concentration of Atorvastatin were measured by HPLC-UV method after beenextracted by ethyl acetate, progesterone was used as internal control.Probability-probability plot and Quantile-quantile plot test were usedto analyze the normality of blood concentration.3.The levels of totalcholesterol(TC),low density lipoprotein cholesterol(LDL-C),high densitylipoprotein cholesterol(HDL-C), triglyceride(TG) in serum of thepatients were analized by automatic biochemical analyzer with the methodof homogeneous oxidation,the samples were collected from the patientsbefore therapy and continued treatment4weeks later.The Q-Q plot testwas used to inspect the normality of the data for lipid concentration.4.The patients were divided into three teams according to the genotype,wild-type homozygote, mutant heterozygote and mutant homozygote.Thedifferences of atorvastatin blood concentration and lipid-adjustingefficacy for different genotypes were analized by ANOVA one-way test withSPSS13.0software. SNK-q test was used in the comparison of inter-teams.Pearson correlation analysis was used to find out the correlation betweenblood concentrations of atorvastatin and change rate of lipid-adjusting,p＜0.05was considered as statistically significant difference.[RESULTS]1. The genotypes’distributions of CYP3A4*18A, CYP3A4*18B andMDR1C3435T for the115patients were consistent with Hardy-Weinberggenetic equilibrium (p＞0.05).The mutation frequency of CYP3A4*18A was3.48%, genotype frequency of TT was94.78%, TC was3.48%, CC was1.74%;mutation frequency of CYP3A4*18B was23.48%,GG genotype frequency was60.0%, GA was33.04%, AA was6.96%; mutation frequency of MDR1C3435T was31.74%,CC genotype frequency was46.96%,CT was42.61%, TT was10.43%.2.The overall level of ATV concentration was4.87±2.11ng mL-1(x±s).For CYP3A4*18A,ATV plasma concentration for the patientswith TT genotype was4.79±2.57ng mL-1, the patients with TC genotype was5.06±1.26ng mL-1,CC genotype was5.72±2.84ng mL-1.For MDR1C3435T gene,ATV plasma concentration for the patients with CC genotype was4.81±1.99ng mL-1,CT genotype was4.65±2.88ng mL-1,TT genotype was5.67±3.83ng mL-1.ATV concentration showed no significant differences forthe three genotype teams of CYP3A4*18A or MDR1C3435T(p＞0.05).For theCYP3A4*18B gene,ATV concentration for the patients with GG genotype was4.36±2.39ng mL-1, GA genotype was5.09±2.16ng mL-1,the data of patientswith AA genotype was7.06±4.17ng mL-1,ATV concentrations of homozygousmutant group were significantly higher than those of heterozygous andhomozygous wild type group(F=4.289,p=0.016＜0.05),the latter two groupshad no significant difference(p=0.395>0.05).3.The change of serum lipidin the three genotype teams of CYP3A4*18A and MDR1C3435T showed nosignificant differences(p＞0.05).The efficacy to total cholesterol forthe CYP3A4*18B homozygous mutant patients was significant better than theother two genotype teams(p＜0.05),but the efficacy for patients withheterozygous and wild homozygous of CYP3A4*18B showed no significantdifference(p＞0.05).The change rate of LDL-C for the three genotype teamsof CYP3A4*18B showed significantly differences,from high to low efficacyteams were mutant homozygous, mutant heterozygous and wild typehomozygous (Both p＜0.05). The efficacy to TG and HDL,three genotype teamsof CYP3A4*18B showed no significant differences(p＞0.05).4.There wassignificant correlation between the plasma concentration of ATV and thechange rate of total cholesterol(p=0.031＜0.05).The change rate of TG,LDL-C and HDL-C were not correlated with the ATV plasma concentrationsignificantly(p＞0.05).[Conclusions]1.The frequency of homozygous wild-type(CC genotype) forMDR1C3435T is46.96%,mutant heterozygous （CT genotype）is42.61%,mutanthomozygous(TT genotype) is10.43%in the population of hyperlipidemia,the mutation frequeny of MDR1C3435T is31.74%.The frequency of homozygouswild-type(TT genotype) for CYP3A4*18A is94.78%,mutant heterozygous （TC genotype）is3.48%,mutant homozygous(CC genotype) is1.74%in thepopulation of hyperlipidemia, the mutation frequeny of CYP3A4*18A is3.48%. The frequency of homozygous wild-type(GG genotype) for CYP3A4*18Bis60.0%,mutant heterozygous（GA genotype）is33.04%,mutant homozygous(AAgenotype) is6.96%in the population of hyperlipidemia, the mutationfrequeny of CYP3A4*18A is3.48%.2.The SNPs of MDR1C3435T and CYP3A4*18Ado not affect the plasma concentration of atorvastatin,but the plasmaconcentration of atorvastatin for the man who carries CYP3A4*18B gene ismore than the others. The plasma concentration of ATV and it,slipid-adjusting efficacy is positively correlated within a certainrange.3.The SNPs of MDR1C3435T and CYP3A4*18A do not affect atorvastatin,slipid-adjusting efficacy,the efficacy of atorvastatin for the patientscarry the CYP3A4*18B are more significant than the others. |
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