葡萄糖醛酸转移酶UGT2B7-A268G基因多态性对癫痫儿童丙戊酸血药浓度的影响

目的：在丙戊酸（VPA）单药治疗的癫痫儿童中评估葡萄糖醛酸转移酶UGT2B7-A268G位点的遗传基因多态性对VPA血清浓度的影响。方法：本研究纳入200例癫痫患儿的丙戊酸血药浓度,测定VPA稳态血清浓度。对UGT2B7编码区的A268G采用聚合酶链反应（RPLF）扩增进行基因鉴定分型。根据UGT2B7基因多态性分析VPA血清药物浓度与基因多态性的关系。结果：携带变异UGT2B7-268G一个基因型或纯合基因患儿的VPA血清药物浓度显著高于携带AA基因的患儿。由于儿童个体差异较大,根据年龄、体质量调整VPA浓度后与基因多态性仍然显著关联（P<0.05）。UGT2B7-A268G的基因多态性与Hardy-Weinberg平衡一致（P>0.05）,其中UGT2B7-268A>G等位基因频率分布的是30.00%,而G突变的基因分布频率为70.00%。结论：癫痫患儿UGT2B7基因的A268G突变可能改变丙戊酸的药物代谢动力学过程,并且不受年龄、体质量等因素干扰。UGT2B7的基因多态性对儿童丙戊酸血药浓度产生影响,测定其基因型对于获得适当的丙戊酸稳态浓度和设定起始用药剂量有积极意义     

汉族癫痫患者UGT1A6基因多态性对丙戊酸血药浓度的影响

摘 要 目的:检测尿苷二磷酸葡萄糖醛酸基转移酶（UGT）1A6 A541G基因多态性在黑龙江汉族癫痫患者中的分布频率,探讨UGT1A6基因多态性对丙戊酸血药浓度的影响。方法: 应用荧光偏振免疫法（FPIA）测定患者丙戊酸钠的血药浓度,PCR-RFLP技术检测患者UGT1A6 541位点的基因多态性,PCR扩增产物进行测序验证方法的可靠性。结果:97例汉族癫痫患者中有28名（28.9%）携带突变型UGT1A6基因,其中服用单位剂量（mg·kg^-1）引起的血药浓度AA基因型患者为（3.06±0.80）μg·ml^-1,AG基因型患者为（2.21±0.66）μg·ml^-1,两者差异有统计学意义（P<0.05）。结论:UGT1A6基因多态性对丙戊酸血药浓度有影响,临床上对于含突变型UGT1A6基因的患者服用丙戊酸时应较常规减少用药减量。     

UGT基因多态性对丙戊酸药动学影响研究

癫痫是慢性反复发作性短暂脑功能失调综合征,是神经系统常见疾病之一。丙戊酸(valproate acid,VPA)是一线抗癫痫药物,具有疗效好、起效快、复发率低等优点。但在临床应用中个体差异较大。丙戊酸在体内大部分经尿苷二磷酸葡萄糖醛酸基转移酶(UDP-glucuronosyltransferase,UGT)代谢。现综述UGT基因多态性对丙戊酸代谢影响的研究情况,为临床丙戊酸的个体化用药提供参考。     

UGT1A6、UGT1A9基因多态性对汉族癫痫患者丙戊酸血药浓度的影响

目的:考察尿苷二磷酸葡萄糖醛酸转移酶(UGT)1A6和1A9基因多态性对汉族癫痫患者丙戊酸血药浓度的影响。方法:选取2014年1月—2015年4月于我院门诊就诊的汉族癫痫患者107例,均使用丙戊酸单药治疗,治疗时间为3个月~6年。采用酶放大免疫法测定患者体内丙戊酸稳态血药浓度,采用基质辅助激光解吸电离飞行时间质谱法检测其UGT1A6(rs2070959、rs6759892)和UGT1A9(rs13418420、rs2741045、rs2741049、rs6731242、rs72551330)基因型,并考察基因多态性与丙戊酸标准化血药浓度(CDR)的相关性。结果:未检出UGT1A9 rs72551330突变型,其余6个位点基因型频率均符合Hardy-Weinberg平衡(P>0.05)。UGT1A6 rs2070959、rs6759892突变基因携带(AG+GG或TG+GG型)者丙戊酸CDR值显著低于其野生纯合子携带(AA或TT型)者,差异均有统计学意义(P<0.05);而携带UGT1A9 rs13418420、rs2741045、rs2741049和rs6731242野生纯合子和突变基因的患者丙戊酸CDR值比较,差异均无统计学意义(P>0.05)。结论:汉族癫痫患者UGT1A6 rs2070959、rs6759892基因多态性与丙戊酸血药浓度有关,且UGT1A6 rs2070959、rs6759892突变基因携带者可能需要更高的丙戊酸剂量。     

CYP2C19基因多态性对精神分裂症患者丙戊酸血药浓度的影响

目的 研究CYP2C19基因多态性与精神分裂症患者心境稳定剂丙戊酸血药浓度的关系。方法 选择奥氮平治疗且服用丙戊酸钠作为心境稳定剂的精神分裂症患者160例,采集血液,测定CYP2C19基因型以及丙戊酸血药浓度,比较各个基因型血药浓度的差异。结果 *2/*2型(122.06±41.30)mg·L^-1和*2/*3型(132.34±51.34)mg·L^-1患者丙戊酸血药浓度明显高于野生*1/*1型(79.41±25.14)mg·L^-1(P<0.05或P<0.01);*1/*1、*1/*2和*1/*3型之间、*2/*2和*2/*3型之间血药浓度无统计学差异;PM型血药浓度(122.13±42.85)mg·L^-1与EM型(80.59±48.60)mg·L^-1比较,显著升高(P<0.05)。 结论 *2/*2和*2/*3型CYP2C19患者,其血液丙戊酸浓度较高,该类患者服用丙戊酸作为精神分裂症心境稳定剂时,宜适当降低用药剂量。     

UGT1A6(T19G)基因多态性对神经外科预防使用丙戊酸钠血药溶度的影响

目的:探讨UGT1A6(T19G)基因多态性对丙戊酸钠血药浓度的影响.方法:回顾性分析2020年1月—2020年12月我院神经外科围手术期使用丙戊酸钠患者55例,对其行UGT1A6(T19G)基因检测及丙戊酸血药浓度监测,评估UGT1A6(T19G)基因多态性对丙戊酸血药浓度的影响.结果:UGT1A6(T19G)基因频...     

UGT2B7基因多态性与抗结核药物性肝损伤的相关性分析

目的 探讨UGT2B7基因多态性与抗结核药物性肝损伤(ADIH)易感性之间的关系.方法 在2012年9月到2013年3月唐山市结核病医院确诊并接受规范抗结核治疗的结核病患者中选取发生肝损伤的182例结核病患者为病例组,未发生肝损伤的193例患者为对照组.对两组患者进行流行病学调查、提取DNA、鉴定基因型并进行统计学分析.结果 UGT2B7基因第268位点突变型杂合子AG和802位点突变型杂纯合子TT携带者发生抗结核药致肝损伤的危险性高于野生型携带者,差异有统计学意义(P＜0.01);在调整了个别不相关因素的影响后,UGT2B7 268和802位点的基因多态性与抗结核药致肝损伤的发生仍显著相关.结论 UGT2B7的-268和802位点基因多态性与抗结核药物性肝损伤具有相关性.     

癫痫患者CYP2D6*10基因多态性对丙戊酸钠血药浓度的影响

目的:比较不同基因型与血药浓度之间的关系,为实行个体化用药方案提供参考。方法:采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)法检测癫痫患者CYP2D6*10的基因型,高效液相色谱法测定丙戊酸钠的血药浓度。结果:CC、CT、TT3种基因型基因分别占20.9%、24.4%、54.7%;3种基因型癫痫患者服用丙戊酸钠达稳态血药浓度后测得的血药浓度、标准化血药浓度之间差异无统计学意义(P>0.05)。结论:CYP2D6*10基因多态性对癫痫患者服用丙戊酸钠的血药浓度没有影响。     

Plasma level of AcMPAG in renal transplant recipients is related to the polymorphism of UGT2B7 211G 〉 T

AIM： To clarify whether Mycophenolic acid（MPA） or its metabolite AcMPAG can cause gastrointestinal disturbances, and to explore the effect of UGT2B7 SNP 211G 〉 T on the pharmacokinetic of AcMPAG. METHODS： Twenty-four renal transplant patients were enrolled in this study. Pharmacokinetic study was performed on day 14 after transplantation and symptoms were recorded on the same day. Multiple blood samples were collected before dosing and 0.5,1,1.5,2,4,6,8, 10 and 12 hours after morning dosing. Plasma concentrations of MPA, MPAG and AcMPAG were detected by HPLC. Genotype of UGT2B7 211G 〉 T was determined using PCR-RFLP method. RESULTS： No significant difference was observed between patients with and without side effects for AUC（0-12） of MPA and MPAG. The values of AUC（0-12）/dose of MPA, MPAG and AcMPAG were （39.7 ± 12.3）,[第一段]     

UGT1A9 C-2152T和UGT2B7 G211T突变在中国汉族人群中的分布

目的：建立Touch-down PCR/RFLP的方法检测UGT1A9C-2152T突变,建立PCR/RFLP的方法检测UGT2B7G211T突变,确定其在中国汉族人群中的突变频率。方法：采用Touch-down PCR/RFLP方法,对100名无亲缘关系的汉族男性志愿者进行UGT1A9C-2152T的基因分型。采用PCR/RFLP方法,对363名无亲缘关系的汉族志愿者（其中男性263名、女性100名）进行UGT2B7 G211T的基因分型。结果：在100名中国汉族男性受试者中,末发现UGT1A9C-2152T的突变,与亚洲人通过测序报道的结果基本一致。在363名汉族受试者中,UGT2B7G211T突变发生频率为0.158,与日本人通过测序报道的结果基本一致。中国男性和女性的等位基因频率分别为0.128和0.110,男性的突变频率比女性高（χ^2=6.784,P=0.034）。结论：用PCR/RFLP的方法对UGT2B7 G211T突变分型的方法简便、快速、重复性好,可用于大样本人群的基因检测。UGT2B7G211T突变在中国汉族人中发生频率较高。     

UGT1A9C-2152T和UGT2B7G211T突变在中国汉族人群中的分布

目的建立Touch-down PCR/RFLP的方法检测UGT1A9 C-2152T突变,建立PCR/RFLP的方法检测UGT2B7 G211T突变,确定其在中国汉族人群中的突变频率.方法采用Touch-down PCR/RFLP方法,对100名无亲缘关系的汉族男性志愿者进行UGT1A9 C-2152T的基因分型.采用PCR/RFLP方法,对363名无亲缘关系的汉族志愿者(其中男性263名、女性100名)进行UGT2B7 G211T的基因分型.结果在100名中国汉族男性受试者中,末发现UGT1A9 C-2152T的突变,与亚洲人通过测序报道的结果基本一致.在363名汉族受试者中,UGT2B7 G211T突变发生频率为 0.158,与日本人通过测序报道的结果基本一致.中国男性和女性的等位基因频率分别为 0.128 和 0.110,男性的突变频率比女性高(χ2=6.784, P=0.034).结论用PCR/RFLP的方法对UGT2B7 G211T突变分型的方法简便、快速、重复性好,可用于大样本人群的基因检测.UGT2B7 G211T突变在中国汉族人中发生频率较高.     

Influence of UGT1A8 and UGT2B7 genetic polymorphisms on mycophenolic acid pharmacokinetics in Japanese renal transplant recipients

Objective UGT1A8 and UGT2B7 are important uridine diphosphate-glucuronosyltransferase isoforms for the glucuronidation of mycophenolic acid (MPA). The aim of this investigation was to elucidate MPA pharmacokinetics in UGT1A8 and UGT2B7 genotypes in Japanese renal transplant recipients. Methods Seventy-two recipients received repeated doses of mycophenolate mofetil and tacrolimus. On day 28 after renal transplantation, plasma MPA concentrations were measured for the next 24 h using high-performance liquid chromatography. UGT1A8*2 (A¹⁷³G) and UGT2B7*2 (Y²⁶⁸) were detected using a PCR-RFLP-based procedure. Results There were no significant differences in daytime and nighttime pharmacokinetics of MPA between UGT1A8 or UGT2B7 genotypes. The mean daytime dose-adjusted AUC_0–12 of MPA in UGT1A8*1/*1, *1/*2 and *2/*2 were 2.47, 2.33 and 2.57 ng·h/ml/mg/kg (P = 0.7711), and the mean nighttime AUC_0–12 were 2.15, 2.00 and 2.08 ng·h/ml/mg/kg (P = 0.4656). The mean daytime and nighttime dose-adjusted AUC_0–12 of MPA in UGT2B7*1/*1, *1/*2 and *2/*2 were 2.61, 2.24 and 2.03 ng·h/ml/mg/kg and 2.18, 1.94, and 1.45 ng·h/ml/mg/kg, respectively (P = 0.3475 and 0.2575). The mean nighttime C_max, t_max, and AUC_6–12/AUC_0–12 ratio (enterohepatic circulation and recirculation ratio) of MPA in all UGT1A8 and UGT2B7 genotypes were lower, longer, and higher, respectively, than the daytime values. Conclusions Both UGT1A8 and UGT2B7 allelic variants seem not to affect Japanese interindividual variability for plasma MPA concentration. Regardless of UGT1A8 and UGT2B7 genetic polymorphisms, the absorption of MPA through enterohepatic recirculation is higher at night.     

焦磷酸测序技术检测UGT1A3和UGT2B7在中国汉族人群中的基因多态性

目的建立焦磷酸测序技术(pyrosequencing)研究二相代谢酶UGT1A3和UGT2B7基因多态性在中国汉族人群中的分布。方法应用带生物素标记扩增引物并经PCR扩增和Beads分离,制备UGT1A3和UGT2B7焦磷酸测序单倍摸板。在PYroMarkID焦磷酸测序上进行焦磷酸测序,检测233血样的DNA标本的17个SNP位点,以确定血样DNA标本的的基因型。结果 233例血样的DNA标本中,UGT1A3等位基因有9种表型,分别为UGT1A3*1*1、UGT1A3*1*2、UGT1A3*1*3、UGT1A3*1*4、UGT1A3*1*5、UGT1A3*2*3、UGT1A3*2*4、UGT1A3*3*3和UGT1A3*3*5。UGT2B7-1和UGT2B7-2各有3种基因型,分别为G/G型、G/T型、T/T和C/C型、C/T型、T/T型。结论我国汉族人群中UGT1A3和UGT2B7基因突变较高。     

Limited influence of UGT1A1*28 and no effect of UGT2B7*2 polymorphisms on UGT1A1 or UGT2B7 activities and protein expression in human liver microsomes

AIMS: UGT1A1 and UGT2B7 are enzymes that commonly contribute to drug glucuronidation. Since genetic factors have been suggested to contribute to variability in activities and expression levels of these enzymes, a quantitative assessment of the influence of the major genotypes (UGT1A1*28 or UGT2B7*2) on enzyme activities was conducted. METHODS: Using a bank of microsomal samples from 59 human livers, the effect of UGT1A1*28 or UGT2B7*2 polymorphisms were investigated on rates of estradiol 3-glucuronidation (a marker of UGT1A1 enzyme activity) or zidovudine glucuronidation (a marker of UGT2B7 enzyme activity) and levels of immunoreactive protein for each enzyme. Glucuronidation rates for both enzymes were measured at K(m)/S(50) and 10 times K(m)/S(50) concentrations. RESULTS: UGT1A1 and UGT2B7 enzyme activities varied up to 16-fold and sixfold, respectively. Rates at K(m)/S(50) concentration closely correlated with rates at 10 times K(m)/S(50) concentration for both enzymes (but not at 1/10th K(m) for UGT2B7). Enzyme activities correlated with relative levels of immunoreactive protein for UGT1A1 and UGT2B7. Furthermore, rates of zidovudine glucuronidation correlated well with rates of glucuronidation of the UGT2B7 substrate gemcabene, but did not correlate with UGT1A1 enzyme activities. For the UGT1A1*28 polymorphism, consistent with levels of UGT1A1 immunoreactive protein, mean UGT1A1 activity was 2.5- and 3.2-fold lower for TA(6)/TA(7) (P < 0.05) and TA(7)/TA(7) (P < 0.001) genotypes in comparison with the TA(6)/TA(6) genotype. CONCLUSIONS: Relative to the observed 16-fold variability in UGT1A1 activity, these data indicate only a partial (approximately 40%) contribution of the UGT1A1*28 polymorphism to variability of interindividual differences in UGT1A1 enzyme activity. For the UGT2B7*2 polymorphism, genotype had no influence on immunoreactive UGT2B7 protein or the rate of 3'-azido-3'-deoxythymidine glucuronidation.     

Amino terminal domains of human UDP-glucuronosyltransferases (UGT) 2B7 and 2B15 associated with substrate selectivity and autoactivation

Despite the important role of UDP-glucuronosyltransferases (UGT) in the metabolism of drugs, environmental chemicals and endogenous compounds, the structural features of these enzymes responsible for substrate binding and selectivity remain poorly understood. Since UGT2B7 and UGT2B15 exhibit distinct, but overlapping, substrate selectivities, UGT2B7-UGT2B15 chimeras were constructed here to identify substrate binding domains. A UGT2B7-15-7 chimera that incorporated amino acids 61-194 of UGT2B15 glucuronidated the UGT2B15 substrates testosterone and phenolphthalein, but not the UGT2B7 substrates zidovudine and 11alpha-hydroxyprogesterone. Derived apparent K(m) values for testosterone and phenolphthalein glucuronidation by UGT2B7-15((61-194))-7 were similar in magnitude to those determined for UGT2B15. Moreover, glucuronidation of the non-selective substrate 4-methylumbelliferone (4MU) by UGT2B7-15((61-194))-7 and UGT2B15 followed Michaelis-Menten and weak substrate inhibition kinetics, respectively, whereas 4MU glucuronidation by UGT2B7 exhibited sigmoidal kinetics characteristic of autoactivation. Six UGT2B7-15-7 chimeras that incorporated smaller domains of UGT2B15 were subsequently generated. Of these, UGT2B7-15((61-157))-7, UGT2B7-15((91-157))-7 and UGT2B7-15((61-91))-7 glucuronidated 4MU, but activity towards the other substrates investigated here was not detected. Like UGT2B7, the UGT2B7-15((61-157))-7, UGT2B7-15((91-157))-7 and UGT2B7-15((61-91))-7 chimeras exhibited sigmoidal 4MU glucuronidation kinetics. The sigmoidal 4MU kinetic data were well modelled using both the Hill equation and the expression for a two-site model that assumes the simultaneous binding of two substrate molecules at equivalent sites. It may be concluded that residues 61-194 of UGT2B15 are responsible for substrate binding and for conferring the unique substrate selectivity of UGT2B15, while residues 158-194 of UGT2B7 appear to facilitate the binding of multiple 4MU molecules within the active site.     

不同丙戊酸盐和丙戊酸剂型对丙戊酸血药浓度的影响

目的：研究不同丙戊酸盐和丙戊酸剂型对丙戊酸血药浓度的影响。方法：回顾性分析某院270例丙戊酸血药浓度监测报告,记录患者姓名、年龄、体质量、丙戊酸的用法与用量、联合用药情况（合用药品及其用法与用量）和血药浓度监测结果。分析不同丙戊酸盐和丙戊酸剂型对丙戊酸标准血药浓度的影响。结果：二元Logistic回归分析结果表明丙戊酸盐类型和剂型对标准血药浓度有显著影响（P<0.05）;丙戊酸镁的标准血药浓度[（9.18±3.54）μg·kg·mL^-1·mg^-1]大于丙戊酸钠盐的标准血药浓度[（6.76±2.54）μg·kg·mL^-1·mg^-1];丙戊酸缓释片的标准血药浓度[（8.38±3.49）μg·kg·mL^-1·mg^-1]大于丙戊酸普通片的标准血药浓度[（6.88±2.54）μg·kg·mL^-1·mg^-1],差异均具有显著性（P<0.05）。结论：不同丙戊酸盐和丙戊酸剂型对丙戊酸血药浓度存在明显影响,如何选择丙戊酸盐和丙戊酸剂型对丙戊酸的合理运用具有重要意义。     

Glucuronidation of 1'-hydroxyestragole (1'-HE) by human UDP-glucuronosyltransferases UGT2B7 and UGT1A9.

Estragole (4-allyl-1-methoxybenzene) is a naturally occurring food flavoring agent found in basil, fennel, bay leaves, and other spices. Estragole and its metabolite, 1'-hydroxyestragole (1'-HE), are hepatocarcinogens in rodent models. Recent studies from our laboratory have shown that glucuronidation of 1'-HE is a major detoxification pathway for estragole and 1'-HE, accounting for as much as 30% of urinary metabolites of estragole in rodents. Therefore, this study was designed to investigate the glucuronidation of 1'-HE in human liver microsomes in vitro and identify the specific uridine diphosphate glucuronosyltransferase (UGT) isoforms responsible for 1'-HE glucuronidation. The formation of the glucuronide of 1'-HE (1'-HEG) followed atypical kinetics, and the data best fit to a Hill equation, resulting in apparent kinetic parameters of Km = 1.45 mM, Vmax = 164.5 pmoles/min/mg protein, and n = 1.4. There was a significant intersubject variation in 1'-HE glucuronidation in 27 human liver samples, with a CV of 42%. A screen of cDNA expressed UGT isoforms indicated that UGT2B7 (83.94 +/- 0.188 pmols/min/mg), UGT1A9 (51.36 +/- 0.72 pmoles/min/mg), and UGT2B15 (8.18 +/- 0.037 pmoles/min/mg) were responsible for 1'-HEG formation. Glucuronidation of 1'-HE was not detected in cells expressing UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, and UGT1A10. 1'-HE glucuronidation in 27 individual human liver samples significantly (p < 0.05) correlated with the glucuronidation of other UGT2B7 substrates (morphine and ibuprofen). These results imply that concomitant chronic intake of therapeutic drugs and dietary components that are UGT2B7 and/or UGT1A9 substrates may interfere with estragole metabolism. Our results also have toxicogenetic significance, as UGT2B7 is polymorphic and could potentially result in genetic differences in glucuronidation of 1'-HE and, hence, toxicity of estragole.     

癫痫患者丙戊酸与拉莫三嗪药代动力学相互作用研究

目的 考察癫痫患者合用丙戊酸(VPA)与拉莫三嗪(LTG)的药代动力学相互作用.方法 将纳入的癫痫患者随机分为3组:VPA组276例、LTG组254例和合用组334例.VPA组患者每天口服VPA 500～1000 mg,LTG组每天口服LTG 100～200 mg,合用组用VPA+LTG.高效液相色谱-质谱联用法测定V...