Honokiol attenuates acetaminophen-induced acute liver injury by inhibiting hepatic CYP1A2 activity and improving liver mitochondrial dysfunction / 中草药·英文版

OBJECTIVE@#Acetaminophen (APAP) overdose is a common cause of liver injury. This study aimed to investigate the protective effect of honokiol (Hon) against APAP-induced hepatotoxicity and its potential mechanism.@*METHODS@#C57BL/6 mice were administrated with Hon (10 and 30 mg/kg) after APAP (300 mg/kg) treatment. On 1.5 h and 5 h after Hon treatment, mice were sacrificed. Serum and liver were collected. And then, liver injury-related indexes, APAP metabolism-related indexes, mitochondrial respiratory chain function-related indexes, and mitochondrial membrane function-related protein expression were evaluated.@*RESULTS@#It was found that Hon significantly decreased serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) activity and glutathione (GSH) depletion, increased hepatic catalase (CAT) and GSH peroxidase (GSH-Px) activities, reduced hepatic MDA and 3-nitrotyrosine contents, inhibited hepatic CYP1A2 activity and APAP protein adducts (APAP-CYS) formation. Meanwhile, oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV in mitochondrial respiratory chain was increased, whereas the release of H2O2 in the mitochondria was decreased following Hon treatment. Furthermore, Hon markedly down-regulated p-JNK in both cytosol and mitochondria, and obviously inhibited the release of apoptosis inducing factor (AIF) and endonuclease G (EndoG) from mitochondria to cytosol.@*CONCLUSION@#Hon alleviated APAP-induced liver injury through the following pathways: Reducing the production of APAP-CYS by inhibiting CYP1A2 activity; Ameliorating hepatic oxidative stress by increasing the levels of hepatic CAT, GSH-Px and GSH; Improving mitochondrial respiratory chain function by promoting oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV; Improving the function of mitochondrial membrane by inhibiting p-JNK and its translocation to mitochondria, thereby reducing the release of AIF and EndoG.

Molecular Docking of Volatile Oily Constituents of Chinese Herbal Medicine Asari Radix et Rhizoma and CYP1A2 Enzyme / 中国实验方剂学杂志

Objective::To study the mechanisms of action of four volatile oil components (safrole, myristicin, methyleugenol and asarone) and the reactive metabolites of safrole and myristicin with CYP1A2. Method::The inhibitory effects of the volatile oil components of Asari Radix et Rhizoma on the human liver microsomal enzymes CYP1A2, CYP2D6, CYP2E1, CYP3A4 and CYP2C19 were screened by the " Cocktail" probe substrate method. The ability of the volatile oil components and intermediates in binding to CYP1A2 enzyme was studied by means of semi-flexible molecular docking. Result::The screening results showed that the components had a strong inhibitory effect on CYP1A2.Molecular docking scores were 3.048 7 kcal·mol-1 (safrole), 6.016 4 kcal·mol-1 (myristicin), 16.969 2 kcal·mol-1 (methyleugenol), 16.013 8 kcal·mol-1 (asarone), 23.923 3 kcal·mol-1 (safrole reactive metabolites) and 25.594 3 kcal·mol-1 (myristicin reactive metabolites). Conclusion::Molecular docking results indicate that safrole metabolic intermediate and myristicin metabolic intermediate have the strongest ability in binding to CYP1A2 enzyme. This study further confirms that safrole and myristicin are the mechanism-based inhibitors of CYP1A2 enzyme, which is consistent with the results of previous IC50-shift and glutathione capture experiments.

Effects of CYP1A2 Gene Polymorphisms on Blood Concentrations of Antipsychotic Drugs ：A Meta-analysis / 中国药房

OBJECTIVE：To systematically evaluate the effects of CYP1A2 gene polymorphisms on blood concentrations of antipsychotic drugs （haloperidol，clozapine，olanzapine），and to provide evidence-based reference for the clinical drug use . METHODS：Retrieved from Cochrane Library ，PubMed，Embase，CBM，CNKI and Wanfang data ，during the inception to Nov. 2019，cross-sectional study was conducted to investigate the effects of CYP1A2 gene polymorphisms on blood concentrations of antipsychotic drugs （haloperidol，clozapine，olanzapine）were collected. After screening the literature ，extracting the data and quality evaluation with Q-Genie tool ，Meta-analysis was performed by using Rev Man 5.3 software. RESULTS ：A total of 11 cross-sectional studies were included ，with a total of 914 patients. Of these ，haloperidol was used in 2 studies，clozapine was used in 5 studies，and olanzapine was used in 4 studies. Meta-analysis showed that there was no statistically significant difference in the blood concentration of haloperidol between CYP1A2（－2964G＞A）G/G type and G/A+A/A type [SMD ＝－0.22，95％CI（－0.66， 0.23），P＝0.35]. The blood concentration of clozapine in CYP1A2（－163C＞A）A/C type was significantly lower than C/C type [SMD ＝0.31，95％CI（0.01，0.62），P＝0.04]；there was no statistical significance in blood concentration of clozapine between A/A type and C/C type [SMD ＝0.09，95％CI（－0.21，0.40），P＝0.56]，between A/A type and A/C type [SMD ＝－0.22，95％CI（－0.55， 0.10），P＝0.18]，between CYP1A2（－2467delT）delT/delT type and T/T type [SMD ＝－0.11，95％CI（－0.75，0.52），P＝0.72]， between delT/T type and T/T type [SMD ＝0.01，95％CI（－0.33，0.34），P＝0.97]，between delT/delT type and delT/T type [SMD ＝ －0.15，95％CI（－0.80，0.05），P＝0.66]. The blood concentration of olanzapine in CYP1A2（－163C＞A）A/A type was signifi- cantly lower than A/C type [SMD ＝－0.31，95％CI（－0.55， － 0.08），P＝0.009]；there was no statistically significant difference in the blood concentration of olanzapine between A/A liukefeng-num.1@163.com type and C/C type [SMD ＝－0.20，95％CI（－0.61，0.21），P＝ 0.34]，between A/C type and C/C type [SMD ＝0.06，95％CI E-mail：hnmuzj@163.com （－0.35，0.47），P＝0.77]，between CYP1A2（－2467delT）delT/T type and T/T type [SMD ＝0.28，95％CI（－0.15，0.71），P＝0.20]. CONCLUSIONS：CYP1A2（－163C＞A）A/C type is related to the reduction of clozapine blood concentration ，and A/A type is related to the reduction of olanzapine blood concentration. CYP1A2 （－163C＞A）gene polymorphism is significance for guiding individualized medication of schizophrenia patients.

Detoxification mechanism of Aconiti Lateralis Radix Praeparata combined with dried Rehmanniae Radix based on metabolic enzymes in liver / 中国中药杂志

The enzymes CYP1 A2 and CYP3 A4 were measured by building a "Cocktail" probe drug and the incubation system of liver microsomes. The compatibility of Aconiti Lateralis Radix Praeparata combined with dried Rehmanniae Radix on CYP450 enzyme protein and gene expression was explored from the level of protein and molecular biology. It explored the molecular mechanism of compatibility detoxication of Aconiti Lateralis Radix Praeparata to provide scientific support for clinical safe and effective application of Aconiti Lateralis Radix Praeparata. The CYP450 enzyme activity was determined by using "Cocktail" probe drugs. The content of CYP450 enzyme was measured by CO reduction of differential spectrum method. The mRNA expression of CYP1 A2 and CYP3 A4 enzyme was detected by RT-PCR technology. Compared with the blank group, the CYP1 A2 and CYP3 A4 enzyme activity and mRNA expression were increased in the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group with significant differences(P<0.05), while the CYP3 A4 enzyme activity and mRNA expression were no influence in the Aconiti Lateralis Radix Praeparata group. The CYP3 A4 enzyme activity and mRNA expression were increased in the dried Rehmanniae Radix and the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group, and there were significant differences(P<0.05). The content of CYP450 enzyme was decreased in the Aconiti Lateralis Radix Praeparata group, contributed to extremely significant difference(P<0.01). The content of CYP450 enzyme was increased in the dried Rehmanniae Radix and the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group, and there were significant differences(P<0.05). The CYP1 A2 and CYP3 A4 enzyme activity and gene expression were enhanced after dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata. The metabolism of toxic ingredients of Aconiti Lateralis Radix Praeparata was accelerated to reach an effect of detoxication. The detoxication mechanism of compatibility of Aconiti Lateralis Radix Praeparata was verified from the viewpoint of liver metabolic enzymes.

Muscle strength and caffeine supplementation: are we doing more of the same? / Força muscular e suplementação de cafeína: estamos fazendo mais do mesmo? / Fuerza muscular y suplementación de cafeína: ¿estamos haciendo más del mismo?

ABSTRACT The purpose of this review was to examine in the current literature the advances made in terms of the effects of caffeine supplementation on maximum strength and its associated mechanisms since the publication of two important papers in 2010. Searches were carried out in the PubMed, Medline, Scielo and Web of Science databases for articles published after 2010. Sixteen studies were included based on inclusion and exclusion criteria. Five studies did not report changes in maximal voluntary strength (31.3%). Four of them used isometric muscle contractions, although this may not be a key factor because five other studies also used isometric contractions and reported ergogenic effects. Furthermore, these four studies evaluated small muscle groups and volunteers were not accustomed to consuming caffeine. Caffeine produced ergogenic effects in eleven of the sixteen studies analyzed (68.8%). None of the doses were clearly related to ergogenic effects; however, a dose of at least 3 mg/kg of caffeine is probably necessary. Caffeine ergogenicity was affected by various factors. There was a lack of standardized protocols and controls for intervening factors (e.g., circadian cycles and nutritional states), which could affect results. An ideal caffeine supplementation protocol that is useful for future research, athletes, and physical activity practitioners, has yet to be defined. A small advance made since 2010 involved a possible lack of gender difference; it would appear that caffeine supplementation affects men and women equally. Level of Evidence I; Systematic Review of Level I Studies.

RESUMO O objetivo desta revisão foi examinar na literatura atual os avanços feitos com relação aos efeitos da suplementação de cafeína sobre a força máxima e seus mecanismos associados a partir de 2010, ano em foram publicados dois importantes artigos. As buscas foram realizadas nas bases de dados PubMed, Medline, Scielo e Web of Science procurando-se artigos publicados após 2010. Dezesseis estudos foram incluídos com base nos critérios de inclusão e exclusão. Cinco estudos não relataram alterações da força voluntária máxima (31,3%). Quatro deles usaram contrações musculares isométricas, embora isso possa não ser um fator chave, porque outros cinco estudos também usaram contrações isométricas e relataram efeitos ergogênicos. Além disso, esses quatro estudos avaliaram pequenos grupos musculares e os voluntários não eram habituados à cafeína. A cafeína produziu efeitos ergogênicos em 11 dos 16 estudos analisados (68,8%). Nenhuma dose foi claramente relacionada com efeitos ergogênicos; contudo, há indícios da necessidade de uma dose de pelo menos 3 mg/kg de cafeína. A ergogenicidade da cafeína foi afetada por vários fatores. Havia uma falta de protocolos padronizados e controle de fatores intervenientes (por exemplo, ciclo circadiano e estado nutricional) que poderiam afetar os resultados. Ainda é preciso definir um protocolo ideal de suplementação de cafeína que seja útil para futuras pesquisas, atletas e praticantes de atividade física. Um pequeno avanço feito desde 2010 envolveu a possível falta de diferença de gênero - parece que a suplementação de cafeína afeta homens e mulheres igualmente. Nível de Evidência I; Revisão Sistemática de Estudos de Nível I.

RESUMEN El objetivo de esta revisión fue examinar en la literatura actuallos avances hechos con respecto a los efectos del suplemento de cafeína sobre la fuerza máxima y sus mecanismos asociados desde 2010, año en que se publicaron dos artículos importantes. Las búsquedas se realizaron en las bases de datos PubMed, Medline, Scielo y Web of Science, por artículos publicados después de 2010. Se incluyeron 16 estudios basados en los criterios de inclusión y exclusión. Cinco estudios no reportaron cambios de la fuerza voluntaria máxima (31,3%). Cuatro de ellos usaron contracciones musculares isométricas, aunque esto puede no ser un factor clave, porque otros cinco estudios también utilizaron contracciones isométricas y reportaron efectos ergogénicos. Además, estos cuatro estudios evaluaron grupos musculares pequeños y los voluntarios no tenían el hábito de consumo de cafeína. La cafeína produjo efectos ergogénicos en 11 de los 16 estudios analizados (68,8%). Ninguna dosis fue claramente relacionada con efectos ergogénicos, sin embargo, hay indicios de la necesidad de una dosis de al menos 3 mg/kg de cafeína. La ergogenicidad de la cafeína se vio afectada por varios factores. Hubo una falta de protocolos y controles estandarizados de factores intervinientes (por ejemplo, ciclo circadiano y estado nutricional) que podrían afectar los resultados. Todavía es necesario definir un protocolo ideal de suplemento de cafeína que sea útil para futuras investigaciones, atletas y practicantes de actividad física. Un pequeño avance hecho desde 2010 involucró la posible falta de diferencia de género - parece que el suplemento de cafeína afecta a hombres y mujeres igualmente. Nivel de Evidencia I; Revisión sistemática de estudios de Nivel I.

Compound Compatibility Law of Rhubarb and Aconite Decoction Based on CYP450 Enzymes / 中国药学杂志

OBJECTIVE: To study the law of compatibility and detoxification of rhubarb and aconite decoction based on the CYP450 enzyme-mediated metabolic interaction. METHODS: The activities of CYP1A2 and CYP3A4 enzymes were determined by incubating the "Cocktail" probe drugs in vitro. The total content of CYP450 enzyme in liver microsomes was determined by carbon monoxide differential method. And the expression of CYP1A2 and CYP3A4 mRNA was detected by RT-PCR method. RESULTS: Rhubarb group, aconite root combined with rhubarb group and rhubarb combined with asarum group showed significant induction effects on CYP1A2 enzyme activity. CYP1A2 enzyme activity was significantly inhibited in the aconite root combined with asarum group. CYP3A4 enzyme activity was significantly inhibited in the asarum group and the aconite root combined with asarum group. Rhubarb group, rhubarb combined with asarum group and rhubarb and aconite decoction had significant induction effects on CYP3A4 enzyme activity. Rhubarb group significantly induced the total content of CYP450 enzyme, asarum group and aconitum root combined with asarum group inhibited the total content of CYP450 enzyme, rhubarb and aconite decoction had slight induction effects on the total content of CYP450, but there was no significant difference. Rhubarb group, rhubarb combined with asarum group and rhubarb and aconite decoction group could up-regulate the mRNA expression of CYP1A2. In addition, rhubarb and aconite decoction and rhubarb group could up-regulate the mRNA expression of CYP3A4. And asarum group and aconite combined with asarum group could down-regulate the mRNA expression of CYP3A4. CONCLUSION: The drug combination weakened the strong induction of CYP1A2 and CYP3A4 enzymes by rhubarb alone, reflecting the holism concept of compound traditional Chinese medicine. The effects of rhubarb and aconite decoction on CYP3A4 enzyme activity are likely to be regulated by the mRNA levels of CYP3A4 enzyme. Whether there is correlation between the cold-heat compatibility based on the pharmacological theory and the induction or inhibition of CYP450 enzyme needs further study.

Effects of aqueous extract of Polygoni Multiflori Radix and its main constituents on expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells / 中草药

Objective To analyze the main chemical constituents and their contents in aqueous extract of Polygoni Multiflori Radix (PMR, root of Polygonum multiflorum), and to elucidate the effects of aqueous extract of PMR and its main constituents on the expression of the mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells. Methods The main chemical constituents and their content in aqueous extract of PMR were determined by HPLC. The cytotoxicity of aqueous extract of PMR and its main constituents on L02 cells was determined by MTT assay. The mRNA expression of CYP1A2, CYP2C9, and CYP2E1 in L02 cells were detected by quantitative real-time PCR. Results There were four main well-separated chromatographic peaks standing for tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside, emodin and physcion in aqueous extract of PMR. The content of thesecomponents in aqueous extract of PMR was (1.14 ± 0.03)%, (0.106 9 ± 0.001 6)%, (0.010 8 ± 0.000 9)%, (0.003 55 ± 0.000 19)%, respectively. The cytotoxicity of aqueous extract of PMR and emodin on L02 cells at 24 h was dose-dependent, and the concentration of 50% inhibition was 7.290 mg/mL and 0.082 mmol/L respectively. Tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside and physcion did not show significant cytotoxicity on L02 cells in the experimental concentrations. Aqueous extract of PMR and emodin significantly inhibited the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells. Emodin-8-O-β-D-glucoside inhibited the expression of mRNA of CYP1A2 and CYP2C9. Tetrahydroxy stilbene glucoside inhibited the expression of mRNA of CYP1A2 but activated the expression of mRNA of CYP2C9. Physcion inhibited the expression of mRNA of CYP1A2 and CYP2C9 in a dose-dependent manner, but inhibited the expression of mRNA of CYP2E1 in low concentration and activitated the expression of mRNA of CYP2E1 in high concentration. Conclusion The inhibition of aqueous extract of PMR on the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells is the combined effect of all components in it. The main four components all inhibit the expression of mRNA of CYP1A2. The anthraquinone is the main component inhibiting the expression of mRNA of CYP2C9. The free anthraquinone is the main component inhibiting the expression of mRNA of CYP2E1.

Effect of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos before and after compatibility on activities of CYP1A2, CYP2E1, and CYP3A4 from rat liver microsomes / 药物评价研究

Objective To study the influence of Salviae Miltiorrhizae Radix et Rhizome (SMR) and Carthami Flos (CF) before and after compatibility on activitis of cytochrome P 1A2 (CYP1A2),cytochrome P2E1 (CYP2E1),and cytochrome P3A4 (CYP3A4) from rat liver microsomes.Methods Using caffeine,chlorzoxazone,and midazolam as the probe drugs ofCYP1A2,CYP2E1,and CYP3A4,the SD rats were randomized divided into four groups:control group,SMR (1.2 g crude drug/kg) group,CF (0.4 g crude drug/kg) group,and SMR (1.2 g crude drug/kg) + CF (0.4 g crude drug/kg) group.According to the above dose,rats were ig given drugs for 7 d.Rats were injected with caffeine,chlorzoxazone,and midazolam solution in tail vein 30 rain after the last administration,and the blood was collected at different time points.Metronidazole as internal standard,method has been established to determine the levels of caffeine,chlorzoxazone,and midazolam to evaluate the activities of CYP1A2,CYP2E1,and CYP3A4 by HPLC.Results Compared with control group,SMR increased the clearance rates (CL) of caffeine,chlorzoxazone,and midazolam,reduced the AUC,and t1/2 was also show a decreasing trend,but the difference was not significant.In CF group,CL of caffeine and chlorzoxazone was decreased,but the difference is not significant.CL of midazolam significantly decreased (P ＜ 0.01).AUC of chlorzoxazone increased,but the difference was not significant.AUC of caffeine and midazolam increased significantly (P ＜ 0.05 and 0.01).In SMR + CF group,the CL of caffeine and chlorzoxazone decreased significantly (P ＜ 0.05),the AUC of caffeine and chlorzoxazone increased significantly (P ＜ 0.05),and t1/2 also showed a decreasing trend,but the difference is not significant.Conclusion Compatibility of SMR and CF has an inhibitive effect on CYP1A2 and CYP2E1 in rats,and it could be one of the mechanisms ofinteractive synergy.

Fatty acid participates in up-regulation of diabetes on function and expression of CYP1A2 / 中国药理学通报

Aim To investigate the mechanism of dia-betes changing the hepatic CYP1 A2 through in vitro cell culture study.Methods The function of CYP1 A2 in HepG2 and Fa2N-4 cells were evaluated by determi-ning the level of phenacetin metabolism,and the mR-NA expression of CYP1 A2 in cells was detected by real time PCR.HepG2 cells were co-cultured with serum of diabetic rats(type 1 and type 2)and normal rats,then the CYP1 A2 function in cells were evaluated.Then, the HepG2 and Fa2N-4 cells were co-cultured with a series of concentrations of saturated (including palmitic acid and stearic acid)and unsaturated fatty acids(in-cluding oleic acid and linoleic acid)for 48 h,and the function and expression of CYP1 A2 in the cells were compared.Results It was found that the activities of CYP1 A2 were higher in cells incubated with diabetic serum of both type.All high concentration of fatty acids could increase the function and expression of CYP1 A2 in both HepG2 and Fa2N-4 cells.Conclusion It is speculated that the abnormal level of fatty acids under diabetic state might be part of the reasons why diabetes change the hepatic CYP1 A2,which provides the basis for future study.

Effect of Shenmai Iniection on isoforms of cytochrome P450 enzymes in rats / 中草药

Objective: To investigate the effects of Shenmai Injection (SMI) on activities of six isoforms of cytochrome P450 (CYP450) by Cocktail probe drugs in rats. Methods: SD rats were randomly divided into SM group and blank control group, which were given SMI (10 mL/kg) or normal saline for 7 d. Phenacetin, bupropion, tolbutamide, omeprazole, metoprolol, and midazolam were used as probe drugs for CYP1A2, CYP2B1, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. The UPLC-MS/MS method was used to determine the concentration of probe drugs in rat plasma, and the pharmacokinetic parameters were estimated by DAS3.0. Results: Compared with the blank control group, AUC0~∞, CL, and Cmax of phenacetin, bupropion, and omeprazole were significantly decreased (P < 0.05). However, no significant difference of plasma concentration and pharmacokinetics for tolbutamide, metoprolol, and midazolam was shown between SMI group and the blank control group. Conclusion: SMI can inhibit CYPlA2, CYP2B1, and CYP2C19 activities significantly, but has no effect on the activities of CYP2C9, CYP2D6, and CYP3A4.

Effects of Ziziphus jujuba fruit extracts on cytochrome P450 (CYP1A2) activity in rats / 中国天然药物

Drug-drug interactions have become a serious problem in the clinic, since plant-based medicines are extensively used. The present study investigated the effects of Ziziphus jujuba fruit (ZJ) extract on the pharmacokinetics of phenacetin, a typical substrate of a cytochrome P450 enzyme CYP 1A2, in rats. The rats were pretreated with the water extract (1.0 g · kg(-1)) or the ethanolic extract (3.6 g · kg(-1)) of ZJ for 10 days, and the pharmacokinetics of phenacetin was investigated after intravenous administration. In an in vitro assay, acetaminophen formation in the hepatic microsomes of ZJ-treated rats was investigated to assess CYP1A2 activity. Our results demonstrated that the treatment with the water and ethanolic extracts of ZJ decreased the plasma concentration of phenacetin and increased the plasma concentration of acetaminophen, resulting in a 43.2% and 15.5% reduction in the AUC0-120 of phenacetin, respectively, and a 53.2% and 64.9% increase in the AUC0-120 of acetaminophen, respectively after intravenous administration. The water or ethanolic extract of ZJ significantly increased the clearance of phenacetin and acetaminophen formation in hepatic microsomes. In conclusion, ZJ extracts displayed effects on the pharmacokinetics of phenacetin and increased the CYP1A2 activity in rats. Therefore, precaution on drug-drug interactions should be taken when ZJ is co-administered with drugs metabolized by CYP1A2, which may result in decreased concentrations of these drugs.

Determination of CYP1A2 and CYP2D1 activities in rat liver microsomes and kinetic studies using ultra performance liquid chromatography tandem mass spectrometry / 中国药学杂志

OBJECTIVE: To establish a robust, fast and convenient method for in vitro assay of rat liver CYP1A2 and CYP2D1, and explore their kinetic features. METHODS: Two selective substrates including phenacetin and dextromethorphan, which are probes of CYP1A2 and CYP2D1, were chosen for liver microsomes incubation, respectively; the corresponding ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS) methods were developed for kinetic studies. RESULTS: The fast and convenient UPLC-MS methods with high resolution and short running time (4-5 min) were established and validated for two assays of CYP1A2 and CYP2D1 activities; both methods showed good accuracy and precision, and the values of LOQ for CYP1A2 and CYP2D1 assays could reach 0.267 and 0.007 μmol·L-1, respectively. The kinetic studies showed that the Michaelis constant (Km) for CYP1A2 and CYP2D1 were (28.4±2.7) and (13.9±1.3) μmol·L-1, respectively. Their activities were determined to be (1.47±0.12) and (3.98±0.09) nmol·mg-1, respectively, when the substrate concentration was 10 μmol·L-1. CONCLUSION: UPLC Tandem MS technique is proved to be a rapid, convenient and efficient approach with high sensitivity and selectivity for the assays of CYP1A2 and CYP2D1 in drug metabolism.

Association of CYP1A2 and CYP2D6 gene polymorphism with therapeutic efficacy of duloxetine on depression in southern region of Fujian / 中华行为医学与脑科学杂志

Objective To explore the influence of CYP1A2 and CYP2D6 gene polymorphism on blood concentration,therapeutic efficacy and adverse effects of anti-depression drug duloxetine on depression patients in southern region of Fujian.Methods 82 patients with depression were selected from southern region of FuJian, China,and all participates received duloxetine for two weeks.Blood concentrations of duloxetine were detected by HPLC-MS,and CYP1A2 and CYP2D6 genotypes were determined by sequencing with the amplified PCR products from peripheral blood DNA.The therapeutic efficacy and adverse effects of duloxetine were evaluated by Hamilton depression scale (HAMD) and treatment emergent symptom scale (TESS) respectively.Results Subjects were divided into GG,GA and AA three groups based on CYP1A2 * 1C genotyping.There was no significant difference in blood concentrations of duloxetine, dose-corrected blood concentrations, the reduction rate of HAMD and the reduction rate of TESS among the three groups.Results were the same with CYP1A2 * 1F,which were divided into CC, CA and AA three groups.Subjects was divided into CC, CT,TT three groups based on CYP2D6 * 10 locus genotyping.Blood concentrations of duloxetine were (13.89±3.22) ng · ml-1 , (16.08±4.24) ng · ml-1 , (17.25±4.62) ng · ml-1 respectively and there was significant difference(F=3.21, P＜0.05).CC group was significantly lower than TT group(P＜0.05) , and CT group was lower than TT group but without significant difference (P＞0.05).Dose-corrected blood concentrations were (304.84± 103.76), (368.13± 143.49), (444.50± 195.58) respectively and there was significant difference(F=4.19, P＜0.05), and CC group was significantly lower than TT group (P＜0.05).The reduction rate of HAMD were 0.42±0.11,0.46±0.11,0.52±0.09 respectively and there was significant difference (F =6.29, P＜0.05), and CC and CT group was significantly lower than TT group(P＜0.05).The reduction rate of TESS were 1.14±0.66,1.48±0.69, 1.69±0.69 respectively and there was significant difference(F=3.38, P＜0.05).CC group was significantly lower than TT group(P＜0.05).Conclusion Among the 3 loci studied,only CYP2D6 * 10 locus within CYP2D6 gene can affect blood concentration,efficacy and adverse effects of duloxetine,which indicate that CYP2D6 gene polymorphism may contribute to therapeutic efficacy of duloxetine on depression in southern region of Fujian.

CYP1A2 polymorphism and theophylline clearance in Korean non-smoking asthmatics

BACKGROUND: Theophylline is mainly metabolized by cytochrome P450 (CYP) 1A2 and CYP2E1 which show inter-individual variations. However, the underlying mechanism remains unknown in humans. We investigated the relationship between differences in theophylline clearance and genetic polymorphisms in the CYP1A2 and CYP2E1 gene in 89 Korean asthmatic patients. METHODS: Polymerase chain reaction (PCR) was performed on the 5'-flanking region of those genes. PCR products were directly sequenced and confirmed using the SNaP shot method. We determined whether the detected SNPs affected gene transcription using electrophoretic mobility shift assay (EMSA). Theophylline clearance (mL/kg/h) was assessed by using a Bayesian approach.

Effects of cefetamet hydrochloride injection on activity of CYP1A2, CYP3A4 and CYP2E1 in liver microsomes of rats / 第二军医大学学报

Objective To study the effect of cefetamet hydrochloride injection on the activity of 3 kinds of cytochrome P450 (CYP450) isoforms (CYP1A2, CYP3A4 and CYP2E1) in rat liver microsomes. Methods The SD rats were randomly divided into two groups: control group and cefetamet hydrochloride (CH) group, with each group containing 3 male rats and 3 female rats. The CH group was injected with cefetamet hydrochloride into the tail vein at 50 mg/(kg • d), twice a day for 7 days. A HPLC method was used for simultaneous determination of the production of metabolites and the degradation of the prototype probe substrates of 3 kinds of CYP450 isoforms, so as to evaluate the activity of hepatic CYP450. The analytical column was Diamonsil C18 column (150 mm X 4. 6 mm, 5 Fm), with the flow rate being 1. 0 mL/min. The mobile phase consisted of methanol (0. 1% formic acid) (A)-water (0. 1% formic acid)(B), 0-5 min; 18%A, 5-10 min; 18%-60%A, 10-15 min: 60%A and detected at 247 nm for determination of CYP1A2 activities; methanol (A)-water (0. 02% formic acid)(B), 0-11 min: 40%-60%A and detected at 223 nm for determination of CYP3A4 activities; and methanol (A)-water, 0-10 min: 37%-75%A and detected at 287 nm for determination of CYP2E1 activities. Results Probe substrates and their metabolites showed good linearity within the determining range (r≥0. 999 7). The precision of the method was 0. 05). Conclusion CH injection can significantly induce hepatic microsome CYP3A4 expression in SD rats, but has no induction or inhibition effect on CYP1A2 and CYP2E1, indicating that potential drug-drug interaction might occur when CH injection is coadministered with drugs metabolized by CYP3A4.

Polimorfismo genético relacionado al metabolismo de la aflatoxina y el riesgo de cáncer vesicular en mujeres chilenas / Aflatoxin metabolism-related genetic polymorphisms and the risk of gallbladder cancer in Chilean women

Background/aim: High consumption of red chili pepper has been shown to be a risk factor for gallbladder cancer (GBC) in Chilean women with gallstones (GS). GS are the main cause of GBC, but not all patients with gallstones develop GBC. Since red chili pepper is a widely consumed spice among the Chilean population, the development of GBC in Chilean women cannot be completely explained by the presence of GS and red chili pepper consumption alone. Genetic factors in addition to these and other environmental factors may also be associated with an increased risk of GBC. We aimed to study whether genetic polymorphisms involved in aflatoxin metabolism are associated with the risk of GBC in Chilean women, because we detected aflatoxins B1 and B2 in red chili pepper purchased in Santiago, Chile. Methods: We conducted a hospital-based case-control study whose subjects were 57 patients with GBC, 119 patients with GS, and 70 controls. DNA was extracted from subjects’ blood or paraffin block samples using standard commercial kits. The statuses of the genetic polymorphisms of cytochrome P450 (CYP) 1A2 rs762551 and CYP3A4 rs2740574 were assayed using the TaqMan® SNP Genotyping Assay or the Custom TaqMan® SNP Genotyping Assay, respectively. Results: In the assay for the CYP1A2 polymorphism, of the 57 GBC patients, 23 (40.3 percent) had at least one minor allele (A/C or C/C). However, there were no significant differences in the genotypic or allelic frequencies among the three subject groups. In the assay for the CYP3A4 polymorphism, the minor G/G genotype was not detected in the three groups, and there were no significant differences in the genotypic or allelic frequencies among the three groups. Conclusion: These genetic polymorphisms were not related to the risk of GBC in Chilean women. Further studies including a greater number of controls and cases are needed to confirm this preliminary exploratory result.

Introducción/objetivo: El alto consumo de ají rojo ha demostrado ser un factor de riesgo de cáncer vesicular (CV) en mujeres chilenas con cálculos vesiculares. Los cálculos vesiculares son la causa principal de CV, no obstante, no todos los pacientes con cálculos vesiculares desarrollan CV. Debido a que el ají rojo es una especia ampliamente consumida entre la población chilena, el desarrollo de CV en las mujeres chilenas no puede ser explicado en su totalidad sólo por la presencia de cálculos vesiculares y consumo de ají rojo. Factores genéticos además de éstos y otros factores ambientales, también podrían estar relacionados con un aumento del riesgo de CV. Nuestro objetivo es estudiar si los polimorfismos genéticos involucrados en el metabolismo de la aflatoxina están relacionados con el riesgo de CV en mujeres chilenas, porque detectamos aflatoxinas B1 y B2 en ajíes rojos comprados en Santiago de Chile. Métodos: El estudio caso control, incluyó 57 pacientes con CV, 119 pacientes con cálculos vesiculares, y 70 controles. Se extrajo ADN de la sangre de los sujetos o de bloques de parafina, usando kits comerciales estándar. El estado de los polimorfismos genéticos del citocromo P450 (CYP) 1A2 rs762551 y CYP3A4 rs2740574 fueron estudiados usando el ensayo de genotipo SNP TaqMan® o el ensayo de genotipo SNP Custom TaqMan®, respectivamente. Resultados: En el ensayo para el polimorfismo CYP1A2, de los 57 pacientes con CV, 23 (40,3 por ciento) tuvieron al menos un alelo menor (A/C o C/C). No obstante, no hubo diferencias significativas en las frecuencias genotípicas o alélicas entre los tres grupos. En el ensayo para el polimorfismo CYP3A4, el genotipo menor G/G no fue detectado en los tres grupos, y no hubo diferencias significativas en las frecuencias genotípicas o alélicas entre los tres grupos. Conclusión: Estos polimorfismos genéticos no estaban relacionados con el riesgo de CV en mujeres chilenas...

The effect of lycopene on the total cytochrome P450, CYP1A2 and CYP2E1.

Aim Some carotenoids such as canthaxantin, astaxanthin and beta apo-8’-carotenal were reported to have modulatory effect on the cytochrome P450. The present study was conducted to investigate the effects of lycopene, a nonprovitamin A carotenoid, on microsomal cytochrome P450, CYP1A2 and CYP2E1. Methods Total cytochrome P450 levels, CYP1A2 and CYP2E1-catalyzed reactions (acetanilide 4-hydroxylation and p-nitrophenol hydroxylation) were studied in the liver microsomes of male Sprague Dawley rats. Microsomes were prepared using differential centrifugation combined with calcium aggregation method. Lycopene was orally administered in the dosages of 0, 25, 50 or 100 mg/kgBW/day for 14 days in a repeated fashion. Data were analyzed using ANOVA test. Results Total cytochrome P450 level and acetanilide 4-hydroxylase activity were unaffected by any of the treatments. The CYP2E1 probe enzyme (p-nitrophenol hydroxylase) was signifi cantly reduced by repeated administration of 100 mg/kgBW/day lycopene (7.88 + 2.04 vs 12.26 + 2.77 n mol/min/mg prot). Conclusion The present results suggest that lycopene does not affect the total cytochrome P450 or CYP1A2 activity but it inhibits the activity of CYP2E1 (p-nitrophenol hydroxylase) in the rat.

Cytochrome P450 1A2~* 1C gene polymorphism in Dai and Han nationality volunteers from Dehong autonomous prefecture in Yunnan province / 中华麻醉学杂志

Objective To investigate cytochrome P450 1A2~* 1C gene polymorphism in Dai and Han nationality volunteers from Dehong autonomous prefecture in Yunnan province. Methods One hundred and seventeen Dai and 112 Han nationality volunteers from Dehong autonomous prefecture in Yunnan province were enrolled in this study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed in genotyping analysis. Results There were 45 wild-type homozygotes (G/G), 63 heterozygotes (G/A) and 9 homozygotes among 117 Dai nationality volunteers, while 63 wild-type homozygotes (G/G), 44 heterozygotes (G/A) and 5 homozygots among 112 Han nationality volunteers. There was significant difference in the incidences of the genotypes between the two populations (P<0.05). The distribution of the genotypes of either population was in Hardy-Weinberg equilibrium. The frequency of the allele A in the 1ocus-2964 of CYP1 A2 was 35% (95% CI30%-40%) and 24% (95% CI 20%-30%) respectively in Dai and Han nationality volunteers. There was significant difference in the frequency between two populations (P<0.05). There was also significant difference in the frequency between Dai nationality volunteers and the populations of other regions. Conclusion CYP1A2~*1C gene polymorphism is one of factors of producing individual and racial variation in pharmacology in Dai and Han people from Dehong autonomous prefecture in Yunnan province.

Genetic Association Research between Mutation in CYP1A2 Gene and Erxian Soup Response / 中国中医药信息杂志

Objective To investigate the genetic association between mutation in cytochrome P450 1A2 (CYP1A2) gene and Erxian suop response. Methods Forty-eight patients with male climacteric syndrome of both Yin and Yang deficiency received a treatment of erxian suop with three months. The severity of symptoms and responses to Erxian suop were quantizated by the method of Department of Psychology in Istanbul Bosphorus University and Clinical Research Guidance Principle on New Drug of Traditional Chinese Medicine published by China. A single nucleotide polymorphism (SNP) at position 2964 (G-A) of CYP1A2 gene were identified by polymerase chain reaction (PCR) in order to analyse the relationship between Erxian response and CYP1A2 single nucleotide polymorphism. Results Statistical analysis showed there was no significant difference among genetype and genetype distribution of CYP1A2 G2964A between the patients and control group. The genotype of the 16 patients among 32 effective patients was G/G (56.25%). The genotype of the 11 patients among 16 ineffective patients was G/A (68.75%). There was no significant difference in symptoms, physical sign between the two groups at the pre-treat and post-treat. Also the two groups had no significant difference among the score of major symptoms and hyposymptoms. Conclusion The patients with genotype of G/G have higher efficiency, while with genotype of G/A have lower efficiency among patients with CYP1A2 G2964A gene when treated with erxian soup.

The Association of Neonatal Hyperbilirubinemia with UGT1A1 and CYP1A2 Gene Polymorphism in Korean Neonates / 소아과

PURPOSE: The incidence of nonphysiologic neonatal hyperbilirubinemia is twice as high in East Asians as in whites. Recently, UGT1A1 mutation was found to be a risk factor for neonatal hyperbilirubinemia. In congenitally-jaundiced Gunn rats, which lack expression of UDP-glucuronosyltransferase, alternative pathways can be stimulated by inducers of CYP1A1 and CYP1A2 enzymes. CYP1A2 plays a major role in bilirubin degradation of the alternate pathway. We studied the relationship between UGT1A1 and CYP1A2 gene polymorphism of neonatal hyperbilirubinemia in Koreans. METHODS: Seventy-nine Korean full term neonates who had hyperbilirubinemia(serum bilirubin >12 mg/dL) without obvious causes of jaundice, were analyzed for UGT1A1 and CYP1A2 gene polymorphism; the control group was sixty-eight. We detected the polymorphism of Gly71Arg of UGT1A1 gene by direct sequencing and T2698G of CYP1A2 by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) using MboII and direct sequencing. RESULTS: Allele frequency of Gly71Arg mutation in the hyperbilirubinemia group was 32 percent, which was significantly higher than 11 percent in the control group(P<0.0001). Mutant gene frequency of T2698G was 41.8 percent in patients and 32.3 percent in the control group(P=0.015), but allele frequency was 21 percent in patients and 19 percent in the control group, which was not significantly higher(P=0.706). There was no relationship between mutations of two genes(P=0.635). CONCLUSION: The polymorphism of UGT1A1 gene(Gly71Arg) and CYP1A2 gene(T2698G) was detected in Korean neonatal hyperbilirubinemia. Only polymorphisms of Gly71Arg in UGT1A1 were significantly higher than control group.