CYP3A5和MDR1基因多态性与肝移植病人他克莫司浓度/剂量比的关系

 目的研究肝移植术后患者的细胞色素P4503A5酶(CYP3A5)和多药耐药蛋白(MDR1)基因多态性对他克莫司浓度/剂量比的影响。方法记录患者的体重、他克莫司剂量和血药浓度等指标,采用聚合酶链式反应-限制性内切片段长度多态性(PCR-RFLP)方法对肝移植患者进行基因分型,比较不同基因型患者之间他克莫司的浓度/剂量比。结果CYP3A5*1/*1和*1/*3型患者的他克莫司浓度/剂量比明显低于*3/*3型患者(P<0.01),MDR1的3435和2677位点各基因型分组之间无明显差异(P>0.05)。结论CYP3A5基因*3多态性与他克莫司血药浓度/剂量比具有显著相关性,*1/*1和*1/*3型的患者拟取得相似的血药浓度要比*3/*3型患者服用更高剂量的他克莫司,用药前检测基因型可以更有效地对他克莫司进行剂量调整。     

供体CYP3A5基因多态性影响肝移植术后FK506的个体化用药

 目的探讨肝移植术后口服免疫抑制剂FK506的剂量及其全血谷浓度的个体差异与供体的肝药酶P450 3A5(CYP3A5)基因多态性的关系。方法观察44例接受肝移植的受体在术后1,2周及1月的FK506服药剂量和全血药物谷浓度,并利用PCR-限制性片断长度多态性(PCR-RFLP)方法和DNA直接测序法检测对应供体CYP3A5基因内含子3第6 986位A/G单核苷酸多态性(CYP3A5*3),分析基因多态性与FK506服用剂量及全血谷浓度/剂量比值(C/D)的相关性。结果肝移植术后FK506的口服需药量在个体间存在极大差异,在术后2周及1月,CYP3A5*3/*3基因型患者需要的剂量最小,分别为(0.074±0.042)和(0.084±0.045)mg·kg^-1·d^-1,而C/D比值明显高于*1/*1基因型患者。结论肝移植术后受体FK506服用剂量的个体化差异与供体CYP3A5*1/*3基因多态性密切相关,分析供体CYP3A5*1/*3基因多态性可以为肝移植术后FK506的个体化用药提供可靠的参考指标。     

CYP3A5*3和MDR1 G2677T/A基因多态性对肾移植患者环孢素A血药浓度及疗效的影响

 目的 研究CYP3A5*3和MDR1 G2677T/A基因突变对肾移植术后患者服用环孢素A浓度/剂量×体表面积（C/D′）、不良反应和急性排斥反应的影响。方法 采用聚合酶链反应（PCR）和限制性内切片段长度多态性（RFLP）方法检测141例肾移植患者CYP3A5*3和MDR1 G2677T/A基因型,比较不同基因型患者之间环孢素A的C/D′值、不良反应和急性排斥反应发生率的差异。结果 CYP3A5*1/*1和CYP3A5*1/*3患者术后7、14、30 d的C₀/D′值明显低于CYP3A5*3/*3型患者(P<0.01),组间不良反应和急性排斥反应发生率却无显著性差异(P>0.05);MDR1 2677位点野生型(GG)、突变杂合型(GT﹢GA)、突变纯合型(TT﹢AA﹢AT)3组基因型患者术后7、14、30 d的C₀/D′值及7、30 d的C₂/D′值存在显著性差异(P<0.05),突变型C/D′值高于野生型,3个月内的急性排斥反应发生率低于野生型(P<0.05),组间不良反应发生率无显著性差异(P>0.05)。结论 CYP3A5*3和MDR1 G2677T/A基因多态性与C/D′值显著相关,且MDR1 G2677T/A与肾移植患者的急性排斥反应存在联系,用药前检测两种基因型有助于环孢素A的个体化给药。     

ABCB1和CYP3A5基因多态性对环孢素血药浓度的影响

 目的 研究ABCB1基因和CYP3A5基因多态性与重症肌无力患者环孢素药物浓度的关系。方法 在129名重症肌无力(MG患者中,采用荧光PCR法测定ABCB1 C3435T基因型;RFLP法分析ABCB1 C1236T, ABCB1 G2677A/T;Mismatch RFLP(错配PCR+RFLP法分析CYP3A5*3基因型,并对这4个位点进行连锁分析。收集患者临床资料,对患者环孢素血药浓度进行检测,并对患者遗传学数据和血药浓度等进行分析。结果 连锁分析显示4个SNP中,ABCB1 C1236T,G2677A/T和C3435T之间紧密连锁,ABCB1中最常见的两个单倍体型分别是1236C-2677G-3435C和1236T-2677T-3435T。另外发现CYP3A5*1/*3位点、ABCB1 C1236T位点和ABCB1 G2677T位点的多态性变化对环孢素血药浓度有明显影响,突变型组的血药浓度明显比野生型组高。同时根据ABCB1单倍体型分组,各组之间药物浓度比较都是TT-TT-TT> CT-GT-CT > CC-GG-CC。结论 药物基因学研究对环孢素治疗重症肌无力患者的临床合理用药有指导意义。     

CYP2C9及VKORC1基因多态性对华法林剂量和抗凝效果的影响

 目的 研究中国汉族人性别、年龄、体重、身高、血清白蛋白、细胞色素P450(CYP)2C9和维生素K环氧化物还原酶复合体1(VKORC1)基因型与华法林剂量的关系,为华法林个体化给药方案提供建议。 方法 收集中国汉族健康受试者以及临床使用华法林的患者,记录患者的年龄、性别、体重、身高、白蛋白水平、凝血酶原时间国际标准化比值(PT-INR)、华法林剂量等指标。采用聚合酶链反应-限制性内切酶片段长度多态性(PCR-RFLP)技术检测CYP2C9和VKORC1基因型。 结果 221例健康受试者的CYP2C9基因型检测有200例为*1/*1型,1例为*1/*2型,20例为*1/*3型;VKORC1（-1639G>A）基因型检测有182例为突变纯合子AA型,39例为杂合子GA型,未发现GG型;190例患者的CYP2C9基因型检测有174例为*1/*1型,16例为*1/*3型,未发现*2突变;VKORC1（-1639G>A）基因型检测有163例为AA型,25例为GA型,2例为纯合子GG型。VKORC1（-1 639G>A）AA组,华法林平均剂量明显低于GA+GG组[(2.61±0.84) mg·d-1比(4.44±0.87) mg·d-1],CYP2C9*1/*3基因型组华法林平均剂量明显低于*1/*1型组[(2.29±0.83) mg·d-1比(2.98±1.09) mg·d-1];CYP2C9基因型、VKORC1基因型分别解释了华法林剂量个体差异的4.2%和39.4%,包括年龄、体重、CYP2C9基因型以及VKORC1基因型的多变量模型能解释个体间剂量差异约55.0％（r2=0.55）。 结论 包括年龄、体重、CYP2C9和VKORC1基因型的多变量个体化给药方案对临床提高华法林使用的安全性有潜在的意义。     

CYP2C19 和 MTHFR 基因变异的交互作用 与食管癌易感性的关联研究

〔摘 要〕 目的：探讨细胞色素 P4502C19（CYP2C19）和亚甲基四氢叶酸还原酶（MTHFR）基因变异的交互作用与 食管癌易感性的关系。方法：收集 2017 年 5 月至 2018 年 12 月赣州市人民医院经胃镜和病理检查确诊为新发食管癌患者 100 例,同时随机抽取相同年龄组无肿瘤病史及家族史的一般人群 200 例（男女各 100 例）作为对照,采用序列特异性引 物聚合酶链反应（PCR–SSP）法检测患者和正常对照者的 CYP2C19（*1、*2、*3）和 MTHFR（C677T）基因型分布。 结果：两组研究对象的 MTHFR 基因型及等位基因频率分布比较,差异均无统计学意义（P ＞ 0.05）;食管癌组 EM、IM 占 比均低于正常对照组,差异具有统计学意义（P ＜ 0.05）;PM 占比方面,食管癌组高于正常对照组,差异具有统计学意义 （P ＜ 0.05）;食管癌组的 EM 型 *1/*1 型、IM 型 *1/*2 型基因分布均低于正常对照组,差异具有统计学意义（P ＜ 0.05）; 而在 PM 型中,食管癌组中 *2/*2 型、*2/*3 型基因分布均高于对照组,差异具有统计学意义（P ＜ 0.05）,两组均无 *3/*3 型 基因分布。结论：MTHFR（C677T）突变跟食管癌的发生无关,CYP2C19 基因慢代谢型增加了食管癌发病的危险性, MTHFR（C677T）与 CYP2C19 基因突变无关联性。     

青海撒拉族人群CYP2C19基因多态性研究

 目的 调查青海撒拉族健康人群中CYP2C19基因型及等位基因的分布特征。方法 采用聚合酶链式反应与限制性内切酶片段长度多态性技术（PCR-RFLP）,对99例健康撒拉族人群CYP2C19基因多态性进行分析。结果 撒拉族人群的CYP2C19*1、CYP2C19*2和CYP2C19*3 3种等位基因的发生频率分别为56.6%、29.8%和13.6%;CYP2C19*1/*1、CYP2C19*1/*2、CYP2C19*1/*3、CYP2C19*2/*2和CYP2C19*2/*3基因型出现频率分别为30.3%、36.4%、16.2%、6.1%和11.1%,CYP2C19*3/*3的纯合子基因型在本试验中未发现。结论 青海撒拉族人群 CYP2C19基因分布明显不同于维族、黎族、傣族、白族、畲族、回族和爱尔兰人,而相似于土族、独龙族、苗族、藏族和汉族。
     

CYP3A5和GSTP1基因多态性与多西他赛联合塞替派治疗转移性乳腺癌近期疗效的相关性研究

 目的 获得中国人群药物代谢酶CYP3A5基因和GSTP1基因两个位点的多态性及其与转移性乳腺癌近期疗效的相关性。方法 所有患者均采用多西他赛联合塞替派化疗方案,对患者每2周期疾病控制率（disease control rate, DCR）进行评估。采用基质辅助激光解吸电离飞行时间质谱（atrix assisted laser desorption ionization/ time of flight, MALDI-TOF）确定两个位点的基因型,比较不同基因型与该化疗方案近期疗效的关系。结果 93例转移性乳腺癌患者中具有CYP3A5 A6986G纯合突变型（GG）的DCR（77.8%）,显著高于AA+AG基因型的（57.4%） ( P<0.05);具有GSTP1 A313G突变型（AG+GG）的DCR（81.6%）显著高于野生型（AA）（57.4%）(P<0.05);对两个基因多态性位点的联合分析显示,同时具有GSTP1 AG+GG和CYP3A5 GG基因型的DCR最高,为84.2% (P<0.05)。结论 CYP3A5和GSTP1的基因多态性与化疗疗效有关, GSTP1 A313G突变型（AG+GG）和/或CYP3A5 A6986G纯合突变型（GG）的患者使用多西他赛联合塞替派方案近期化疗疗效最好,可为临床用药提供参考。     

MDR1G2677T/A基因多态性对环孢素药物代谢动力学及药效学影响的系统评价

 目的 系统评价MDR1G2677T/A基因多态性与环孢素药物代谢动力学及药效学的关系。方法 计算机检索Cochrane图书馆、Pubmed、EMBase、Medline、CNKI 等数据库,检索时间截止至2008年10月。收集有关MDR1 G2677T/A基因多态性与环孢素药代动力学及药效关系的研究。采用Revman 5.0软件对符合纳入标准的研究进行Meta分析。结果 共纳入7篇回顾性研究包括844例肾移植患者,其中英文5篇,中文2篇。Meta分析结果表明,GG基因型患者的剂量调整谷浓度明显低于其他基因型患者(P<0.05),同时GG基因型患者的给药后2 h剂量调整浓度及平均日剂量与(TT+TA+AA)基因型患者之间有显著差异(P<0.05),而与(GT+GA)基因型患者之间无显著差异(P>0.05),G2677T/A基因多态性与急性排斥反应发生率之间无统计学意义的相关性(P>0.05)。结论 G2677T/A基因多态性与环孢素处置有显著相关性,但与急性排斥反应发生率没有关联,同时尚需高质量大样本量的前瞻性研究来证实。     

中国汉族和蒙古族健康人药物代谢酶CYP3A4、CYP2C9、CYP2C19、CYP2D6基因多态性分析

 目的 用基因分型技术对中国汉族、蒙古族健康人CYP3A4、CYP2C9、CYP2C19、CYP2D6进行基因多态性分析,并对汉族人和蒙古族人基因表型和基因频率进行比较。方法 聚合酶链反应-限制性片段长度多态性(PCR-RFLP)对CYP3A4、CYP2C9、CYP2C19、CYP2D6进行分型。结果 汉族、蒙古族健康人CYP3A4*5等位基因频率为0, CYP3A4*18等位基因频率分别为0.183 8、0.202 5,CYP2C9*2等位基因频率分别为0.011 0、0.025 3, CYP2C9*13等位基因频率分别为0、0.00 32,CYP2C19*2等位基因频率分别为0.386 0、0.4146, CYP2C19*3等位基因频率分别为0.051 5、0.044 3,CYP2D6*10等位基因频率分别为0.573 5、0.465 2。结论 汉族、蒙古族健康人群CYP3A4*18、CYP2C19*2、CYP2C19*3、CYP2D6*10等位基因频率均没有显著性差异;本试验在汉族、蒙古族健康人中未发现CYP3A4*5等位基因;仅在蒙古族健康受试人群中发现1人为CYP2C9*1/*13基因型;蒙古族CYP2C9*2等位基因频率远小于汉族（P=0.023）。     

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??OBJECTIVE The analyze the results of tacrolimus blood concentrations in patients with myasthenia gravis and the influencing factors, provide references for rational clinical application. METHODS The data of tacrolimus blood concentrations and medical records in patients with myasthenia gravis in hospital were collected and analyzed from January 2013 to December 2015.Tacrolimus whole-blood concentrations were measured by chemiluminescent microparticle immunoassay and CYP3A5^*3 gene polymorphism were detected by digital fluorescence molecule hybridization fluorescence. RESULTS The range of tacrolimus trough concentrations were from 0 to 64.71 ng??mL^-1 and the average of concentration was(5.22??5.80) ng??mL^-1 in 254 cases with 171 myasthenia gravis patients. The results showed that there was a certain correlation between concentration and dosage, and concentration was also related other factors such as the combination of drugs and CYP3A5^*3 gene polymorphism. CONCLUSION The concentration of tacrolimus are affected by many factors, so when tacrolimus is used, multiple factors should be considered comprehensively to achieve individualized treatment.     

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??OBJECTIVE To investigate the interindividual variabilities of plasma concentration and lipid-regulating efficacy of atorvastatin in patients with hyperlipidemia through the genotyping of CYP3A4*18A, *18B and MDR1 C3435T genes.METHODS One hundred and fifteen Chinese Han population with hyperlipidemia were genotyped by the PCR-RFLP (restriction fragment length polymorphism).The steady-state plasma trough concentrations of atorvastatin were measured by high performance liquid chromatography (HPLC)-UV.The levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TG) were monitored by the homogeneous enzyme method before treatment and 1 month after medication.RESULTS The mutation frequencies of CYP3A4*18A,*18B and MDR1 C3435T were 3.48%,23.48% and 31.74% respectively.It shows no statistically significant difference for the SNPs frequencies between the normal population reported and patients selected.Patients with CYP3A4*18B homozygous mutant (AA) showed a significantly higher plasma concentration of ATV compared with the G/A heterozygous mutat patients or the G/G wild-type homozygous (P=0.016).However,no significant difference could be shown in the patients with CYP3A4*18A and MDR1 C3435T genotypes(P??0.05).Neither CYP3A4*18A nor MDR1 C3435T could be shown a significant difference in the lipid lowering efficiency(P>0.05).Patients carrying the homozygous mutant (AA) of the CYP3A4*18B gene showed a significantly higher TC lipid-regulating effect compared with patients with the GA or GG genetic variant (P=0.02). The LDL-C change rates among the three genotype groups were significantly different, with AA group >GA group >GG group (P=0.01) and the regulation of TG and HDL-C for AA,GA or GG was compared without finding any significant difference (P>0.05).The TC changerates and plasma concentration were significantly correlated (P=0.031) before and after treatment,while there was no statistical significance in the correlation of the other three lipid change rates with plasma concentration (P??0.05).CONCLUSION The SNPs MDR1 C3435T and CYP3A4*18A do not affect the plasma concentration and efficacy of ATV. In ATV therapy, patients with the CYP3A4*18B gene exhibit higher plasma concentrations than the non-carriers, and the lipid-lowering efficacy was more pronounced.     

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??OBJECTIVE To investigate associations between CYP3A4/5 and POR single nucleotide polymorphisms(SNPs)and tacrolimus dose-corrected concentrations(??₀/D) in Chinese adult heart transplant recipients, providing individualized dose-adjustment for this population. METHODS A total of 90 Chinese adult heart transplant recipients in the early stage were enrolled. CYP3A4*1G G??A(rs2242480) genotype was assessed by pyrophosphate sequencing. CYP3A5*3 A>G(rs776746) and POR*28 C>T(rs1057868) genotype were determined by Sanger sequencing. Tacrolimus trough concentration(??₀) was evaluated by enzyme multiplied immunoassay technique(EMIT). Associations between genotypes and ??₀/D as well as time and dose to get the target range were completely analyzed. RESULTS Allele frequencies of all the evaluated SNPs were consistent with Hardy-Weinberg equilibrium (P>0.05). The ??₀/D in CYP3A5*3/*3 carriers was considerably higher than that in *1/*1and *1/*3 carriers. Moreover, time to get the target range was significantly shortened and required dosage was also significantly reduced in CYP3A5*3/*3 carriers. The ??₀/D in CYP3A4*1/*1G carriers was remarkably decreased in comparison with the wild type. After stratification by CYP3A5*3 genotypes, no associations were observed between CYP3A4*1G and POR*28 genotypes and tacrolimus ??₀/D. POR*28 was not related to ??₀/D, but significantly prolonged time to target range. CONCLUSION This study demonstrats that CYP3A4*1G and CYP3A5*3 polymorphisms are associated with tacrolimus concentrations, the test of these genotypes before transplantation may be useful for individualized medicine of tacrolimus.     

ELISA法监测肾移植患者他克莫司全血浓度448例次

 目的 通过监测肾移植后患者他克莫司全血浓度,观察并建立他克莫司在三联免疫抑制用药方案中的理想治疗窗,为临床合理应用提供参考。方法 用ELISA法测定他克莫司全血浓度,对75例患者的448例次监测结果进行比较分析。结果他克莫司全血浓度随移植后时间延长而逐渐下降。肾移植后1个月内、第2～3个月、第4～6个月和>6个月时,用ELISA法监测他克莫司全血谷浓度的治疗窗范围应分别为,8～15,6～12,5～10和3～8μg·L^-1,较为适宜。结论 常规监测他克莫司全血浓度,按推荐治疗窗调整给药方案,可获得满意的免疫抑制治疗效果。     

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??OBJECTIVE To identify the specific cytochrome P450 (CYP) enzymes involved in the metabolism of dipfluzine hydrochloride (Dip) in the rat liver microsomes. METHODS The rat liver microsomes were prepared and incubated with Dip. The Dip metabolites (M1, M2, M4 and M5) were identified by LC-MS/MS, and the CYP isoenzymes were identified by the combination of the selective CYP inhibitor study, correlation analysis and a panel of recombinant rat CYP expereiment, respectively. RESULTS The RESULTS from the experiments of selective CYP inhibitors, correlation analysis and recombinant rat CYP isoenzymes indicated that CYP2A1, CYP3A and CYP2C11 contributed to the formation of M1 and M5 in the rat liver microsomes. CYP3A, CYP2A1, CYP1A2, CYP2C11 and CYP2E1 metabolized Dip to M2. CYP3A, CYP2A1, CYP2E1 and CYP2C11 contributed to M4 formation. And the recombinant rat CYP researches further indicated that CYP3A2 exhibited more activity than CYP3A1. CONCLUSION CYP3A and CYP2A1 are the major CYP isoenzymes responsible for catalyzing Dip to the four metabolites formation in the rat liver microsomes.     

酸枣仁、远志和桔梗水提液对大鼠肝CYP450酶活性 及mRNA表达的调控作用

目的:研究中药酸枣仁、远志和桔梗水提液对P450同工酶在酶活性及mRNA水平的调控作用。方法:大鼠ig给予酸枣仁、远志和桔梗水提液7 d后取肝脏称重并制备肝微粒体,采用紫外分光光度法测定大鼠肝微粒体细胞色素b5(Cytb5),P450的含量及红霉素N-脱甲基酶(ERD)的活性,高效液相色谱法测定非那西汀O-脱乙基酶(PHD)的活性和对硝基苯酚羟化酶(pNPH),采用RT-PCR技术检测大鼠肝中5种P450同工酶CYP1A1,CYP1A2,CYP2E1,CYP3A1及CYP3A2mRNA的表达。结果:与对照组比较,各给药组大鼠肝指数无显著变化;酸枣仁组显著降低Cytb5含量和ERD活性,增加pNPH活性,远志组显著升高pNPH,极显著降低ERD活性,桔梗组显著降低CYP450含量,并明显升高PHD和pNPH活性;在mRNA水平上,各组的CYP1A1基因均未能检出;酸枣仁组诱导CYP2E1,CYP3A1和CYP3A2基因表达,远志组抑制CYP3A1,诱导CYP3A2的mRNA表达,桔梗组诱导CYP1A2,CYP2E1,CYP3A2的mRNA表达;酸枣仁的CYP2E1、桔梗的CYP1A2和CYP2E1mRNA水平基本与酶活性水平相平行。结论:酸枣仁、远志水提液诱导CYP2E1的活性;桔梗水提液诱导CYP1A2,CYP2E1的活性,酶活性变化可能主要通过影响基因转录来实现。提示若酸枣仁和远志与经CYP2E1和CYP3A代谢的药物同用,桔梗若与经CYP1A2,CYP2E1,CYP3A代谢的药物同服,有可能会影响这些药物的临床疗效,临床用药时应慎重考虑。     

Effects of herbal medicinal products and food supplements on induction of CYP1A2, CYP3A4 and MDR1 in the human colon carcinoma cell line LS180

A selection of popular herbal medicinal products and food supplements were analysed for their potential to modulate the expression of the cytochrome P450 enzymes CYP1A2 and CYP3A4 and the transporter protein MDR1. A total of 31 products were analysed. Nine of the products have been approved by the Medical Products Agency (MPA) in Sweden and are marketed as herbal medicinal products. Twenty-two of the products have not been assessed by the MPA and are marketed as food supplements. LS180 cells were exposed to extracts from the different herbal products and real-time quantitative polymerase chain reaction, RT-QPCR, was subsequently used to analyse the relative mRNA levels of CYP1A2, CYP3A4 or MDR1 in treated and non-treated cells. Our results show that 17 of 31 products tested induced a two-fold expression or more for at least one of the genes analysed. Four products, of which a ginger-supplement was the most potent, induced all three genes.