【作者】 邱晓燕； 焦正； 张明； 仲珑瑾； 梁惠琪； 马春来； 张亮； 钟明康；

【Author】 Xiao-yan Qiu1, Zheng Jiao1, 2*, Ming Zhang3, Long-jin Zhong1, Hui-qi Liang1, Chun-lai Ma1, Liang Zhang1, Ming-kang Zhong1 1. Clinical Pharmacy Laboratory, Huashan Hospital, Fudan University, Shanghai, People’s Republic of China 2. School of Pharmacy, Fudan University, Shanghai, People’s Republic of China 3. Department of Nephrology, Huashan Hospital, Fudan University, Shanghai, People’s Republic of China

【机构】 复旦大学附属华山医院临床药学室； 复旦大学附属华山医院肾病科；

【摘要】 目的:探讨中国汉族人中,肾移植患者的多药耐药基因(MDR1)外显子exon12C1236T、exon21G2677T/A、exon26 C3435T及药物代谢酶CYP3A4*18B和CYP3A5*3的单核苷酸多态性(SNP)对免疫抑制剂环孢素A(CsA)药动学的影响。方法:采用聚合酶链反应和限制性内切片段长度多态性(PCR-RFLP)的方法对103例肾移植术后的患者进行MDR1基因分型。CYP3A4*18B和CYP3A5*3的基因分型采用聚合酶链反应和链接酶反应(PCR-LDR)方法。单克隆抗体荧光免疫偏振法测定患者术后CsA的谷浓度(C0)及服药后2h浓度(C2)。比较不同基因型及不同单倍体型之间CsA浓度剂量比值的差异,并以CYP3A4*18B和CYP3A5*3作为分层因素,分层分析MDR1基因多态性对CsA药动学的影响,以消除CYP3A4*18B和CYP3A5*3的混杂因素影响。结果:肾移植术后一个月内,CYP3A5*3基因多态性与CsA的剂量校正C0有相关性。CYP3A5*3/*3型患者,其剂量校正C0在术后8-15天和16-30天,分别比野生型高25.5%(P=0.011),30.7%(P=0.015)。而CYP3A4*18B与CYP3A5*3不同,不仅与C0有关,而且与C2有关。野生型患者的剂量校正C2,在术后8-15天和16-30天比纯合突变型分别提高19.3%(P=0.008)和35.2%(P=0.008),剂量校正C0在术后16-30天比纯合突变型高39.7%(P=0.012)。通过在CYP3A5和CYP3A4表达和不表达的患者中,分别分析MDR1对CsA药动学的影响,以去除CYP3A5和CYP3A4的混杂因素。在CYP3A5和CYP3A4不表达的患者中,MDR1C1236T与CsA剂量校正C0显著相关。在CYP3A5不表达者中,MDR11236CC型患者,其剂量校正C0,在术后1-7天、8-15天和16-30天比TT型患者分别提高37.4%(P=0.011),39.3%(P=0.022)和26.4%(P=0.042)。在CYP3A4不表达者中,其剂量校正C0在术后1-7天、8-15天和16-30天,分别提高30.9%(P=0.016),36.3%(P=0.016)和23.3%(P=0.042)。在所有研究的5种MDR1单倍体中,只有CAC(1236-2677-C3435)单倍体与CsA剂量校正C0有相关性。在CYP3A5不表达者中,CAC单倍体携带患者,其剂量校正C0,在术后1-7天、8-15天和16-30天比非携带者分别提高23.3%(P<0.001),48.6%(P<0.001)和49.2%(P=0.015)。在CYP3A4不表达者中,其剂量校正C0在术后1-7天、8-15天和16-30天,分别提高22.8%(P<0.001),45.3%((P<0.001)和47.3%(P=0.011)。结论:在中国汉族肾移植患者中,CYP3A4*18B的遗传多态性也许能够解释肾移植术后1个月内CsA的药代动力学在个体之间的巨大差异性。携带有CYP3A4*18B等位基因的患者需要较高剂量的CsA已达到目标血药浓度。CYP3A5*3、MDR1C1236TSNPs以及MDR1CAC单倍体只与C0有关,而与C2无关,因此其与肾移植术后CsA的药代动力学的关系,尚需大规模的前瞻性的临床研究证实。更多还原

【Abstract】 Objective:To retrospectively evaluate the effects of MDR1, CYP3A4*18B and CYP3A5*3 genetic polymorphisms on cyclosporine A (CsA) pharmacokinetics in Chinese renal transplant patients during the first month. Methods: A total of 103 renal transplant recipients receiving CsA were genotyped for MDR1 (C1236T, G2677T/A, and C3435T), CYP3A4*18B and CYP3A5*3. The pre-dose and two hour post-dose concentrations of CsA (C0 and C2, respectively) were determined by fluorescence polarization immunoassay, and their relationships with corresponding genotypes and haplotypes were investigated. Results: Patients with a CYP3A4*1/*1 genotype were found to have a higher dose-adjusted concentration compared to those with CYP3A4*18B /*18B, as follows: for C2, 19.3% (P=0.014) during days eight to 15, 35.2% (P=0.008) during days 16 to 30 and for C0, 39.7% (P=0.011) during days 16 to 30. The dose-adjusted C0 was higher in patients with MDR1 1236CC compared to those with 1236TT in the first month post-operation. The dose-adjusted C0 in patients with the CYP3A5*3/*3 genotype was 25.5% and 30.7% higher than those with the wild-type genotype during days eight to 15 (P=0.011) and days 16 to 30 (P=0.015), respectively. Haplotype analysis revealed that the dose-adjusted C0 was higher in the first month following surgery in carriers of haplotype MDR1 CAC than in non-carriers. Polymorphisms of MDR1 and CYP3A5*3 did not affect dose-adjusted C2. Conclusion: The data suggests that the CYP3A4*18B genotype affects CsA pharmacokinetics during the first month following surgery in Chinese renal transplant recipients. Patients with CYP3A4*18B alleles may require higher doses of CsA to reach the target levels. Large prospective studies may be needed to further explore the impact of MDR1 and CYP3A5*3 polymorphisms on CsA pharmacokinetics in renal transplant recipients.更多还原