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Introducción: La lamotrigina (LTG) es un fármaco antiepiléptico aromático, en los humanos se usa principalmente en el campo de la neurología y psiquiatría. Existen eventos adversos (EA), secundarios al uso de anticonvulsivantes, por ejemplo, la necrólisis epidérmica tóxica (NET), y el síndrome de Steven Johnson (SSJ), su incidencia está estimada en 1 de 1000 a 10000 exposiciones con una mortalidad mayor al 35 %. Objetivo: Describir la relación entre la presentación de eventos adversos y los polimorfismos de la UGT y HLA en pacientes que usaron lamotrigina. Metodología: Se realizó una revisión sistemática de la literatura, incluyendo una búsqueda en las bases de datos, con estudios del tipo ensayos clínicos controlados aleatorizados y estudios observacionales de cohortes, casos y controles. Resultados: La búsqueda inicial encontró un total de 96 artículos, de los cuales luego de aplicar los criterios de inclusión y exclusión quedaron 8. Conclusiones: Al momento del desarrollo de este estudio, no hay suficiente evidencia que relacione como factor de causalidad las variantes de las HLA y UGT con la predicción de los eventos adversos (EA), con el uso de lamotrigina. Además, es importante decir que en Colombia no tenemos estudios, y que no conocemos estas variantes genéticas por lo tanto la extrapolación no se puede realizar, llevando a recomendar que se deben hacer estudios en población colombiana con las variantes planteadas como posibles relacionadas con los EA a lamotrigina.
Introduction: Lamotrigine (LTG) is an aromatic antiepileptic drug, mainly used in humans in the field of neurology and psychiatry. There are secondary adverse events (AEs) related to the use of anticonvulsants, for example, toxic epidermal necrolysis (TEN) and Steven Johnson syndrome (SJS). Their incidence is estimated at 1 in 1,000 to 10,000 exposures with a mortality greater than 35%. Objective: To describe the relationship between the presentation of adverse events and UGT and HLA polymorphisms in patients who used lamotrigine. Methodology: A systematic review of the literature was carried out, including a database search, with studies such as randomized controlled clinical trials and observational cohort and case-control studies. Results: The initial search found a total of 96 articles. After applying the inclusion and exclusion criteria, only 8 were considered. Conclusions: At the time of the development of this study, there was not enough evidence to relate HLA and UGT variants as a causal factor to predict adverse events (AEs) due to the use of lamotrigine. Furthermore, it is important to say that in Colombia there are no studies, and that we do not know these genetic variants, therefore it is not possible to extrapolate it. Thus the recommendation is that studies should be carried out in the Colombian population using variants proposed as possibly relating AEs. to lamotrigine.
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Objective:To explore the therapeutic mechanism of Canhuang tablets on the mRNA and protein expression of farnesoid X receptor (FXR), uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) and multidrug resistance associated protein 2 (MRP2) in the liver of jaundiced rats induced by α-naphthalene isothiocyanate (ANIT). Method:The rats were divided into normal group, model group, Canhuang tablets (CHP) group and ursodeoxycholic acid tablets (UDCA) group. The jaundice model was reproduced by ANIT. After the intervention of the corresponding drugs, the contents of total bilirubin (TBIL), total bile acid (TBA), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in serum and the liver histopathology were examined to evaluate the therapeutic effect of CHP. The relative mRNA and protein expressions of FXR, UGT1A1 and MRP2 in rat liver tissues were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Result:CHP can significantly reduce the increase of TBIL, TBA, ALT, AST and ALP caused by ANIT in rat serum, and inhibit the liver pathological changes, which showed that the removing jaundice effect of CHP was better than UDCA. Compared with the normal group, ANIT significantly inhibited the mRNA levels of FXR, UGT1A1 and MRP2 in rat liver tissues after modeling (P<0.01). Compared with the model group, CHP and UDCA significantly increased the mRNA levels of target genes of each protein after intervention (P<0.01), and CHP was superior to UDCA in improving the mRNA level of bilirubin metabolizing enzyme UGT1A1 (P<0.01). In the aspect of affecting protein expression, compared with the normal group, ANIT modeling significantly increased the expression of FXR in rats (P<0.05). CHP intervention showed a tendency to promote the expression of FXR, while UDCA did not, but there was no significant difference between them. In the aspects of promoting bilirubin metabolism and bile excretion, the expressions of UGT1A1 and MRP2 were significantly decreased by ANIT modeling (P<0.01), while the expressions of UGT1A1 and MRP2 proteins were significantly increased after treatment of CHP (P<0.01). CHP was superior to UDCA in increasing the expression of bilirubin and bile acid efflux protein MRP2 (P<0.01). Conclusion:The jaundice abating mechanism of CHP is related to activating FXR mRNA expression in liver, promoting the mRNA and protein expression of bilirubin metabolizing enzyme UGT1A1 and bile acid transporter MRP2, improving liver metabolism of free bilirubin and promoting bile acid excretion from the liver, and alleviating cholestatic liver injury.
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OBJECTIVE: To investigate the metabolic characteristics of bakuchiol mediated by cytochrome P450 enzyme (CYP) and UDP-glucuronosyltransferase (UGT) in rat liver microsomes (RLMs) or human liver microsomes (HLMs), and to compare the metabolic gender differences. METHODS: Bakuchiol was incubated at 37? with male and female RLMs or HLMs in the presence of nicotinamide adenine dinucleotide phosphate (NADPH) or uridine 5′-diphosphoglucuronic acid (UDPGA). The residual concentrations of bakuchiol were measured in each incubation system using high performance liquid chromatography (HPLC). The metabolic stability and metabolic gender differences of bakuchiol were evaluated by the remaining percentage of bakuchiol after incubation.RESULTS: When bakuchiol was metabolized by CYP in RLMs, the intrinsic clearance (Clint) value in male RLMs ?(326.6±15.4) mL·min-1·kg-1?was significantly higher than that of female RLMs ?(77.2±4.8) mL·min-1·kg-1? (P<0.01). When bakuchiol was metabolized by UGT in RLMs, female RLMs had a significantly higher Clint value ?(419.1±24.1) mL·min-1·kg-1? than male RLMs ?(164.5±8.4) mL·min-1·kg-1? (P<0.01). When bakuchiol was metabolized by both CYP and UGT in RLMs, male RLMs had a significantly higher Clint value ?(1063.1±27.2) mL·min-1·kg-1? than female RLMs ?(781.2±16.5) mL·min-1·kg-1?(P<0.01). When bakuchiol was metabolized by CYP in HLMs, male HLMs had a significantly higher Clint value ?(24.8±2.1) mL·min-1·kg-1? than female HLMs ?(17.6±1.0) mL·min-1·kg-1? (P<0.01). There were no significant gender differences in the metabolism of bakuchiol when it was metabolized by UGT in HLMs. The Clint values were 176.4±26.5 and (165.9±8.6) mL·min-1·kg-1, respectively. The metabolic parameters of bakuchiol mediated by CYP and UGT in HLMs had no significant gender differences. The Clint values were 262.5±20.9 and (236.2±10.5) mL·min-1·kg-1, respectively. CONCLUSION Bakuchiol can be metabolized by CYP and UGT in RLMs or HLMs, and the metabolic parameters exhibit species differences and gender differences.
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Objective To investigate the therapeutic effects of emodin on acute cholestatic hepatitis and mec-hanism thereof. Methods Fifty Sprague - Dawley (SD)rats were randomly divided into 5 groups and were treated with emodin,ursodeoxycholic acid,dexamethasone,or normal saline respectively for 4 days. On the fifth day gastric perfusion of alpha - naphthylisothiocyanate(ANIT)was performed to establish models of choiestatic hepatitis. Four to six hours after the establishment of model the above mentioned agents were given continuously. Forty - eight hours after the model establishment blood samples were collected from abdominal aorta to examine the total bilirubin(TB),direct bilirubin (DB),alanine aminotransferase(ALT),total bile acid(TBA),aspartate aminotransferase(AST),alkaline phosphatase (ALP),gamma glutamine transferase(GGT). Specimen of liver was collected to undergo pathological examination. Real - time PCR was used to detect the mRNA expression of farnesoid X receptor(FXR),small heterodimer partner (SHP ),bile salt export pump (BSEP ),uridine diphosphate glucuronosyltransferase 2 family polypeptide B4 (UGT2B4). Results The serum levels of total bilirubin (TB),direct bilirubin (DB),alanine aminotransferase (ALT),total bile acids (TBA),aspartate aminotransferase (AST),and alkaline phosphatase (ALP)of the model group were respectively (68. 1 ± 26. 1)μmol/ L,(46. 3 ± 20. 1)μmol/ L,(483 ± 228)U/ L,(159. 1 ± 57. 9)μmol/L,(2. 0 ± 0. 5)U/ L,(996 ± 382)U/ L,(324 ± 120)U/ L. The levels of TB,DB,ALT,TBA,AST,ALP of the emodin group were respectively (15. 0 ± 8. 7)μmol/ L,(10. 8 ± 3. 9)μmol/ L,(147 ± 71)U/ L,(60. 1 ± 22. 7)μmol/ L, (295 ± 104)U/ L,(222 ± 59)U/ L,and were all significantly lower than those of the model group (all P < 0. 05). The levels of TB,DB,ALT,TBA,AST,GGT,ALP of the emodin group were all significantly lower than those of the ursode-oxycholic acid group (all P < 0. 05). The levels of TB,DB,ALT,TBA,GGT,AST were all significantly lower than those of the dexamethasone group (all P < 0. 01). The expression levels of FXR,SHP,BSEP,UGT2B4 mRNA in the emodin group (1. 087 ± 0. 285,0. 892 ± 0. 390,0. 902 ± 0. 149,1. 785 ± 0. 403)were all significantly higher than those of the model group (0. 152 ±0. 088,0. 559 ±0. 194,0. 561 ±0. 123,0. 177 ±0. 039,all P <0. 05). Conclusions By decreasing the levels of TB,DB,ALT,TBA,AST,ALP and reducing pathological changes,emodin has a protective effect on cholestatic hepatitis. It has better effects than ursodeoxycholic acid and dexamethasone. These effects may be due to promoting FXR,SHP,BSEP and UGT2B4 expression.
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Objective To investigate the relationship between uridine diphosphate glucuronosyltransferases (UGT) 1A6 rs2070959 polymorphism and serum concentration of valproic acid (VPA) in patients with epilepsy.Methods We selected 200 cases of epilepsy patients treated in our hospital from June 2014-January 2017.All the patients were treated with VPA monotherapy > 3months.When the VPA of patients reached steady state,we detected the VPA blood level.The genotypes and allele frequencies of UGT1A6 rs2070959 in 200 epilepsy patients were determined.The average standard deviation of 1-fold VPA was used as high VPA group and vice versa as low VPA group.The genotype and allele frequencies of UGT1A6 rs2070959 were compared between the two groups,and the influencing factors of VPA concentration in epileptic patients were analyzed.Results The frequencies of A genotype,AG genotype and GG genotype were 77.01%,18.39% and 4.60% in the high VPA group,67.26%,19.47% and 13.27% in the low VPA group,with no significant difference between the two groups (P > 0.05);the allele G frequency in the high VPA group was significantly lower than that in the low VPA group (13.79%).The age and weight of the high VPA group were significantly higher than those of the low VPA group (P < 0.05),and the gender composition of the high VPA group was not significantly different from that of the low VPA group (P >0.05).Logistic regression analysis showed that age and weight gain were positively correlated with VPA concentration (P < 0.05),allele G expression was negatively correlated with VPA concentration (P <0.05).Conclusions Allele G expression at UGT1 A6 rs2070959 is associated with decreased VPA concentration in epileptic patients,which may require a higher dose of VPA.
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This study attempts to discuss the correlation between UGT1A1*28 as uridine diphosphate glucuronosyltransferase gene promoter and coding region Gly71Arg gene polymorphism with neonatal hyperbilirubinemia of neonates in Wuhan.A total of 168 neonates were divided into the hyperbilirubinemia group (case group,n=108) and healthy neonates group (control group,n=60).Their DNA was obtained through blood extraction.The gene exon mutation of UGT1A1 was detected by Sanger sequencing,which revealed the relationship between UGT 1A 1*28 and Gly71Arg polymorphism with neonatal hyperbilirubinemia of neonates.The results showed that:(1) The frequency of UGT1Al*28 allele mutation in the case group and the control group was 9.3% and 10% respectively,with the difference being not significant between the two groups (P>0.05).(2) The frequency of Gly71Arg allele mutation in the case group and the control group was 35.1% and 21.7% respectively,with the difference being significant between the two groups (P<0.01).(3) The serum bilirubin level of Gly71Arg mutant homozygous and heterozygous subgroups (n=66) in the case group was 302.7±31.4 μmol/L,which was significantly higher than 267.3±28.5 μmol/L of the wild subgroup (n=42) (P<0.01).It was suggested that the occurrence of neonatal hyperbilirubinemia of neonates in Wuhan was not associated with UGT 1A1*28 gene polymorphism,but closely with the Gly71Arg gene polymorphism.Meanwhile,the Arg allele mutation was related to the degree of jaundice.
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Objective To develop a high-resolution melting ( HRM ) assay for rapidly screening Gilbert syndrome ( GS) and Crigler-Najjar syndrome ( CNS) associated with UGT1A1 defects.Method Methodology was developed .Then, we applied the established method to analyze 61 clinical samples from neonatal patients with severe unexplained unconjugated hyperbilirubinemia .Neonates with known risk factors for developing hyperbilirubinemia , such as ABO hemolysis, G6PD deficiency, sepsis, hypoxic ischemic encephalopathy were excluded .Five pairs of PCR primers were designed to detect the five common mutations (G211A, C686A, C1091T, C1352T and T1456G) in Asia population.PCR and HRM Assay conditions were optimized.UGT1A1 genotyping in clinical samples was performed by using the established HRM analysis , and all results were subsequently confirmed by direct DNA sequencing .Results The mutants were readily differentiated by using HRM analysis .In this study, 42 neonates were identified with UGT1A1 mutation, and 4 different known variants were detected .Conclusion HRM analysis in this study was economical, convenient, rapid, effective for screening UGT1A1 gene mutations, which can serve as an reliable method for the clinical diagnosis of GS and CNS and the large-scale molecular epidemiological research of UGT1A1 gene-related diseases.
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Objective:To investigate the effects and mechanism of Ginseng polysaccharides (GPS) on glycosaminoglycan synthesis in rat chondrocytes in vitro.Methods:Chondrocytes isolated from knee joint cartilage of rats were subcultured.After monolayers were established,chondrocytes induced by 20 ng·ml-1 IL-1β were treated with GPS respectively at the concentration of 2,10 and 50 ng ·ml-1for 48 h.The effects of GPS on chondrocytes were analyzed using MTT assay for chondrocyte proliferation and 1,9-dimethylmethylene blue assay for GAG content.mRNA content in xylosyltransferase-Ⅰ(XylT-Ⅰ),galactosyltransferase-Ⅰ(GalT-Ⅰ),galactosyltransferase Ⅱ(GalT-II) and galactose-β-1,3-glucuronosyltransferase Ⅰ(GlcAT-Ⅰ) and that in matrix-degrading enzymes matrix metalloproteinases-3 (MMP-3) and aggrecanase-1and aggrecanase-2 was also detected.Results:GPS had no toxic or proliferating effect on chondrocytes at all observed concentrations.GPS at the concentrations of 10 ng·ml-1 and 50 ng·ml-1 up-regulated GAG synthesis (P<0.01).10 ng·ml-1 GPS enhanced the mRNA expression of XylT-Ⅰ (P<0.01),and 50 ng·ml-1 GPS enhanced the mRNA expression of XylT-Ⅰ,GalT-Ⅰ and GalT-II (P<0.05,P<0.01).GPS did not inhibit the mRNA expression of matrix-degrading enzymes MMP-3 and aggrecanase-1and aggrecanase-2 enhanced by IL-1β.Conclusion:GPS can improve GAG synthesis in IL-1β stimulated chondrocytes in vitro,which is due to the expression promotion of GAG synthesis enzymes without the expression inhibition of matrix-degrading enzymes.
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La hiperbilirrubinemia no conjugada es una condición producida por una alteración en el proceso de conjugación y excreción de la bilirrubina. La glucoronosiltransferasa uridin difosfato es la responsable en la conjugación de la bilirrubina, es codificada por el gen de UGT1A1 localizado en el brazo q del cromosoma 2 locus 37.1. La variación genética del UGT1A1 puede producir diferentes fenotipos desde el más severo llamado Sindrome Crigler-Najjar Tipo I y II, pasando por el Sindrome de Gilbert; hasta una hiperbilirrubinemia transitoria neonatal o síndrome LUCEY-DRISCOLL (HBLRTFN) fenotipo OMIM 237900 con producción de kernicterus y parálisis cerebral pero con resolución espontánea, todos ellos de herencia autosómica recesiva causada por mutación homocigota o heterocigota en el gen UGT1A1. En este reporte se presenta un caso en un recién nacido que a los 7 días presenta hiperbilirrubinemia severa con kernicterus, y la prueba genética muestra mutación heterocigota del *28 del gen UGT1A1
Unconjugated hyperbilirubinemia is produced by alteration in conjugation and excretion process of bilirubin. Glucoronosiltransferasa Uridine diphosphate enzyme is involved in bilirubin conjugation. Is encoded by the UGT1A1 gene located in chromosome 2q locus 37.1. UGT1A1 genetic variation can produce different phenotypes Crigler-Najjar Syndrome Type I and II, Gilbert Syndrome, and hyperbilirubinemia transited familial LUCEY-DRISCOLL (HBLRTFN) syndrome with kernicterus production but with spontaneous resolution, all autosomal recessive. We present here a case of newborn 7 days old with severe hyperbilirubinemia , kernicterus, and genetic testing shows heterozygous mutation of the UGT1A1 * 28 gene
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Humanos , Vulnerabilidad SexualRESUMEN
Glycyrrhizin is a major bioactive component of liquorice, which exerts multiple biochemical and pharmacological activities and is frequently used in combination with other drugs in the clinic. Mycophenolate mofetil (MMF), an immunosuppressant widely used in transplant patients, is metabolized by UDP-glucuronyltransferases (UGTs). Although significant evidence supports that glycyrrhizin could interact with the cytochrome P450s (CYPs), few studies have addressed its effects on UGTs. The present study aimed at investigating the regulatory effects of diammonium glycyrrhizinate (GLN) on UGTs in vitro and in vivo. We found that long-term administration of GLN in rats induced overall metabolism of MMF, which might be due to the induction of UGT1A protein expression. Hepatic UGT1A activity and UGT1A mRNA and protein expression were significantly increased in GLN-treated rats. UGT1A expression levels were also increased in the intestine, contradicting with the observed decrease in intestinal UGT1A activities. This phenomenon may be attributed to different concentrations of glycyrrhetinic acid (GA) in liver and intestine and the inhibitory effects of GA on UGT1A activity. In conclusion, our study revealed that GLN had multiple effects on the expression and activities of UGT1A isoforms, providing a basis for a better understanding of interactions between GLN and other drugs.
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Animales , Masculino , Ratas , Medicamentos Herbarios Chinos , Química , Farmacología , Glucuronosiltransferasa , Química , Metabolismo , Ácido Glicirrínico , Química , Farmacología , Interacciones de Hierba-Droga , Intestinos , Química , Cinética , Hígado , Química , Ratas Sprague-DawleyRESUMEN
As a bilirubin metabolic disorder, Gilbert syndrome belongs to the category of congenital non-hemolytic jaundice. Deficiency or decrease in the activity of bilirubin-uridine diphosphate glucuronyltransferase (UGT) is an important reason for the pathogenesis of Gilbert syndrome. UGT1A1, an isoenzyme of UGT, is a key enzyme to direct bilirubin in the liver. Mutations in UGT1A1 gene lead to the structural abnormality of UGT, and thus result in the decrease or loss of the ability of UGT to bind bilirubin. This article summarizes the research advances in the role of UGTA1 and its polymorphism in the pathogenesis and diagnosis of Gilbert syndrome.
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This study was designed to investigate the inhibitory effects of sciadopitysin on the catalytic activities of human 12 kinds of UDP-glucuronosyltransferases (UGTs) in vitro. The risk of drug-drug interactions (DDI) is predicted by in vitro-in vivo extrapolation (IV-IVE). Methods A panel of recombinant human UGT isoforms and human liver microsome (HLM) as well as a series substrates including 4-methyl umbelliferone (4-MU), trifluoperazine (TFP) and N-3-carboxypropyl-4-hydroxy-1, 8-naphthalimide (NCHN) (UGT1A1 specific fluorescent probe substrates) were used to characterize the inhibitory effects of sciadopitysin on human UGTs in vitro. The half maximum inhibitory concentration (IC50) and the constant of inhibition kinetics (Ki) were obtained by nonlinear regression using GraphPad Prism 6.0 software. The potential risk of DDI induced by UGT1A1 was predicted based on in vitro parameters. The results demonstrated that sciadopitysin had strong inhibitory effects on UGT1A1, UGT1A3, UGT1A8 and UGT1A10, with the remaining activity being below 30% at a final concentration of 10 μmol·L-1. For UGT1A1, UGT1A3, UGT1A8 and UGT1A10, the IC50 was 0.20 μmol·L-1 to 1.34 μmol·L-1, the inhibition kinetic constant Ki was 0.07 μmol·L-1 to 2.12 μmol·L-1. The AUC ratio of UGT1A1 can be increased by 19% to 147% at the oral dose of 240 mg·d-1. The sciadopitysin competitively inhibited the formation of 4-MU-O-glucuronide by UGT1A1, UGT1A3, UGT1A8, and UGT1A10. At the same time, the inhibition of NCHN-O-glucuronidation by UGT1A1 was consistent with the competitive inhibition. The strong inhibition of sciadopitysin on UGT1A1 led to reduction of the metabolism of UGT1A1 substrates, and increased the risk of DDI. When co-administrated with other drugs, special attentions should be given to the DDI from inhibition of drug metabolism enzymes to prevent serious clinical consequences.
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UDP-glucuronosyltransferase (UGT), a kind of phase II drug-metabolizing enzyme, catalyzes the conjugation of glucuronic acid and drugs. UGTs are widely expressed in brain, but at a relatively low level compared to that in liver. Brain UGTs are inducible or inhibitable, which influences the drug distribution in the central nervous system. UGTs, cytochrome P450 (CYPs) and transporters act together to effect pharmacokinetics of drugs in brain. Several drugs have the capacity to cross the blood brain barrier after glucuronidation and certain drugs may be subject to direct glucuronidate in brain by the function of UGTs. The brain UGTs' isoforms, localization, induction, inhibition, and interaction with CYP and transporters are introduced in this review. The process of drug glucuronidation and distribution in brain is summarized for five drugs. A deep insight into the process of drug metabolism and distribution in brain may provide a valuable reference for drug design for the central nervous system.
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Objective To analyze the relationship between UDP-glucuronosyltransferase 1A1 (UGT1A1)gene polymorphism and delayed diarrhea caused by FOLFIRI treatment.Methods Two hundred and one blood samples were taken from patients with metastatic digestive tract tumor before chemotherapy by FOLFIRI and then the UGT1A1 * 28 genetic polymorphism was performed.All the cases treated with FOLFIRI were chosen to be observed and recorded by situation of the delayed diarrhea during chemotherapy,and to analyze the relationship between UGT1A1 * 28 genetic polymorphism and grade 3 and 4 delayed diarrhea.Results The distributions of the genotypes in 201 metastatic digestive tract tumor patients were as follows:UGT1A1 * 28 wild-type genotype TA6/6 (155,77.11%),heterozygous genotype TA6/7 together with homozygous genotype TA7/7 (46,22.89%).In the 201 cases,the incidences of grade 1 and 2 delayed diarrhea in the patients carrying wild-type genotype and mutant type were respectively 45.16% (70/155),39.13% (18/46).The incidences of grade 3 and 4 delayed diarrhea were respectively 9.68% (15/155),19.57% (9/46),with no statistical difference (x2 =3.318,P =0.190).Conclusion The UGT1A1 * 28 polymorphism TA6/7 or TA7/7 can not increase the risk of grade 3 or more severe delayed diarrhea for the patients with metastatic digestive tract tumor after receiving FOLFIRI treatment.
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Objective To test the mutation locus of uridine diphospho-glucuronosyltransferase gene (UGT1A1) in a Chinese patient with Crigler-Najjar syndrome type Ⅰ and her family members,analyzing the genetic characteristics of the pedigree.Methods Genomic DNA was extracted from the patient and her family members and other 50 full-term infants with normal serum bilirubin as a healthy control group.Fifty cases of full-term newborn whose serum bilirubin level were nomal were study as controls.The promoter and all exons of UGT1A1 gene were amplified by the method of polymerase chain reactions (PCR),and mutations were identified by direct sequencing.Results The propositus and her miscarriage sister were homozygous for a nonsense mutation at nucleotide number 715 (715C > T) in exon 1 of gene UGT1A1,substituting of stop codon (TAG) for glutamine (CAG) at position 239 (Q239X).The other 5 members were heterozygous in the same mutation locus.A TA insertion mutation and a G71R mutation in exon 1 were observed in the family members.The patient and her sister were homozygous of A(TA)7TAA mutation while other four were heterozygous.Propositus,grandmother,mother and her younger brother were heterozygous of G71 R mutation.No mutation was found in exons 2-5.No mutation was found in other fifty healthy cases in the healthy control group.Conclusions Q239X homozygous mutations is considered to be the lethal gene in this Crigler-Najjar syndrome family.Collaborative G71 R and A(TA)7TAA mutations may further reduce the enzyme activity of UGT1A1,causing varying degrees of bilirubin disorder.
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OBJECTIVE: To establish a method to determine the genotypes of the three SNPs(-31T/C, -133C/T and -140T/C) of uridinediphosphateglucuronosyltransferase 1A3(UGT1A3)and to preliminarily analyze the genetic frequencies of different genotypes among healthy Chinese population. METHODS: The genotypes of the three SNPs(-31T/C, -133C/T and -140T/C) of 123 healthy Chinese subjects(89 males, 34 females) were screened by two-step allele specific amplification(two-step ASA) method. Firstly, a pair of primers of the first exon for the amplification of the genomic DNA were designed. And then two parallel reaction tubes were set up for each site which included a forward primer whose last base of 3'end pair with wild type or mutant type and an identical reverse prime for the amplification of the target gene. The amplification Results were examined to differentiate the genotypes. The validity was verified by direct sequencing. RESULTS: Among the 123 subjects, the frequencies of -31T and -31C were 0.776 and 0.224, of -133C and -133T were 0.935 and 0.065, of -140T and -140C were 0.915 and 0.085, respectively. Based on the genotyping Results, there were 59 cases of UGT1A3*1/*1, 1 case of UGT1A3*2/*2, 17 cases of UGT1A3*1/*2, 12 cases of UGT1A3*1/*4, and 1 case of UGT1A3*4/*4. CONCLUSION: There are genetic polymorphisms of UGT1A3 in healthy Chinese population and the mutation frequency is remarkable. The gene detection method established in this study is verified to be sensitive and specific.
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Objective To investigate the role of UDP-glucuronosyltransferase 1A (UGT1A),nuclear factor erythroid-2-related factor 2 (Nrf2) and kelch-like ECH-associated protein 1 ( Keapl ) in the tumorigenesis of colonic carcinoma.Methods The expressions of UGT1 A,Nrf2 and Keapl were detected in normal colonic mucosa(24 cases),adenoma tissue (30 cases) and adenocarcinoma tissue (77 cases) by immunohistochemistry,and the relationship between their expressions and the clinical pathological characteristics was analyzed.Results The positive rates of UGT1 A in normal colonic mucosa,adenoma and adenocarcinoma tissue were 83.3% ( 20/24),80.0% ( 24/30 ) and 53.2% ( 41/77 ),respectively.The positive rate of UGT1A in adenocarcinoma was lower than those in colonic mucosa and adenoma ( all P <0.05 ).On the contrary,the positive rates of Nrf2 in adenoma [70.0% (21/30) ] and adenocarcinoma tissue [ 87.0% (67/77) ] were higher than that in normal colonic tissue [ 41.7% (10/24),all P =0.000 ].The positive rates of Keapl in normal colonic mucosa,adenoma and adenocarcinoma tissue were 54.2% ( 13/24),70.0% (21/30) and 61.0% (47/77),respectively ( normal colonic tissue vs adenocarcinoma tissue,P =0.040 ; adenoma vs adenocarcinoma,P =0.002 ).There was no correlation between the expression of UGT1 A,Nrf2 and the clinicopathologic features of colon carcinoma,while the differences of Keapl positive rates in the various degrees of tumor differentiation [ moderately-well differentiated vs poorly differentiated:70.0% (35/50) vs 44.4% (12/27) ] and invasion [T1-T2 vs T3-T4:78.8% (26/33) vs 47.7% (21/44) ]were statistically significant (all P < 0.05 ).Conclusion The decreased expression of UGT1A and the dysregulation of Nrf2/Keapl system may play a role in colonic tumorigenesis.
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Patients with co-existing hereditary spherocytosis (HS) and UDP-glucuronosyltransferase 1A1 (UGT1A1) deficiency as Gilbert's syndrome (GS) have been reported, and previous studies have demonstrated an increased risk for developing gallstones in patients with co-inheritance of GS and HS. We experienced an interesting case of HS showing persistent jaundice after splenectomy, and upon further evaluation, the 25-year-old female patient was found to have HS combined with UGT1A1 deficiency. Sequence analysis of the UGT1A1 gene revealed that she was a compound heterozygote with p.[G71R; Y486D] + [Y486D] mutations, which suggests Crigler-Najjar syndrome type II rather than GS. Careful evaluation of inappropriately elevated bilirubin level compared with the degree of hemolysis is important, reflecting the therapeutic implication of splenectomy and cholecystectomy.
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Adulto , Femenino , Humanos , Síndrome de Crigler-Najjar/genética , Glucuronosiltransferasa/deficiencia , Heterocigoto , Homocigoto , Ictericia/etiología , Mutación Missense/genética , Mutación Puntual/genética , Esferocitosis Hereditaria/complicaciones , Esplenectomía/efectos adversosRESUMEN
Objective To analyze the relationship between the genetic polymorphisms of uridinediphosphate glucuronosyltransferase 1A9 (UGT1A9) and mycophenolic acid (MPA)pharmacokinetics in Chinese kidney recipients.Methods Gene mutations (C-440T/T-331C,C-2152T,T-275A,T98C) were detected in 196 recipients by PCR-LDR.On the 28th day after transplantation,the plasma samples which were obtained at the time points of predose,0.5 h and 2 h after administration were measured by an immunoassay method (Emit Mycophenolic Acid Assay,Dade Behring).MPA-AUC0-12 was calculated based on these three data.Correlation between single nucleotide polymorphisms (SNPs) and MPA pharmacokinetics was analyzed.Results C-2152T,T-275A and T98C genotypes of UGT1A9 were not found in 196 recipients.The frequency of C-440T/T-331C gene mutation was 14.29% (28/196).The mean value of MPA-AUC0-12 was 40.6±11.8 wild genotype,respectively (P>0.05).Conclusion C-2152T,T-275A and T98C genotypes of UGT1A9 are scarce in Chinese kidney recipients.In this study,there is no distinct relationship between -440/-331 SNPs and MPA pharmacokinetics in Chinese allograft recipients.
RESUMEN
Gilbert's syndrome is a congenital,nonhemolytic,unconjugated hyperbilirubinemia.The intermittent mild jaundice is its clinical characteristic.The pathogenesis of the disease is the gene mutation of the isoenzyme which encoding the UDP-glucuronosyltransferase (UGT1A1).The gene mutation can affect glucuronic acidation of drugs.Therapeutic dose can cause unexpected toxicity.It is very important to detect the gene mutation of UGT1A1 for diagnosis,treatment and genetic counseling of Gilbert's syndrome.