The role of CYP1A1/2 in cholesterol ester accumulation provides a new perspective for the treatment of hypercholesterolemia

Cholesterol is an important precursor of many endogenous molecules. Disruption of cholesterol homeostasis can cause many pathological changes, leading to liver and cardiovascular diseases. CYP1A is widely involved in cholesterol metabolic network, but its exact function has not been fully elucidated. Here, we aim to explore how CYP1A regulates cholesterol homeostasis. Our data showed that CYP1A1/2 knockout (KO) rats presented cholesterol deposition in blood and liver. The serum levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and total cholesterol were significantly increased in KO rats. Further studies found that the lipogenesis pathway (LXRα-SREBP1-SCD1) of KO rats was activated, and the key protein of cholesterol ester hydrolysis (CES1) was inhibited. Importantly, lansoprazole can significantly alleviate rat hepatic lipid deposition in hypercholesterolemia models by inducing CYP1A. Our findings reveal the role of CYP1A as a potential regulator of cholesterol homeostasis and provide a new perspective for the treatment of hypercholesterolemia.

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.

The expression of oxidative stress genes related to myocardial ischemia reperfusion injury in patients with ST-elevation myocardial infarction / 世界急诊医学杂志（英文）

@#BACKGROUND: We aimed to investigate the gene expression of myocardial ischemia/reperfusion injury (MIRI) in patients with acute ST-elevation myocardial infarction (STEMI) using stress and toxicity pathway gene chip technology and try to determine the underlying mechanism. METHODS: The mononuclear cells were separated by ficoll centrifugation, and plasma total antioxidant capacity (T-AOC) was determined by the ferric reducing ability of plasma (FRAP) assay. The expression of toxic oxidative stress genes was determined and verified by oligo gene chip and quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, gene ontology (GO) enrichment analysis was performed on DAVID website to analyze the potential mechanism further. RESULTS: The total numbers of white blood cells (WBC) and neutrophils (N) in the peripheral blood of STEMI patients (the AMI group) were significantly higher than those in the control group (WBC: 11.67±4.85 ×109/L vs. 6.41±0.72 ×109/L, P<0.05; N: 9.27±4.75 ×109/L vs. 3.89±0.81 ×109/ L, P<0.05), and WBCs were significantly associated with creatine kinase-myocardial band (CK-MB) on the first day (Y=8.945+0.018X, P<0.05). In addition, the T-AOC was significantly lower in the AMI group comparing to the control group (12.80±1.79 U/mL vs. 20.48±2.55 U/mL, P<0.05). According to the gene analysis, eight up-regulated differentially expressed genes (DEGs) included GADD45A, PRDX2, HSPD1, DNAJB1, DNAJB2, RAD50, TNFSF6, and TRADD. Four down-regulated DEGs contained CCNG1, CAT, CYP1A1, and ATM. TNFSF6 and CYP1A1 were detected by polymerase chain reaction (PCR) to verify the expression at different time points, and the results showed that TNFSF6 was up-regulated and CYP1A1 was down-regulated as the total expression. GO and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis suggested that the oxidative stress genes mediate MIRI via various ways such as unfolded protein response (UPR) and apoptosis. CONCLUSIONS: WBCs, especially neutrophils, were the critical cells that mediating reperfusion injury. MIRI was regulated by various genes, including oxidative metabolic stress, heat shock, DNA damage and repair, and apoptosis-related genes. The underlying pathway may be associated with UPR and apoptosis, which may be the novel therapeutic target.

Influence of nicotine dependence, current smoking status and gender on CYP1A1, CYP1B1 and CYP2A6 expression in oral squamous cell carcinoma / Influência da dependência nicotínica, parâmetros de consumo de tabaco e gênero na expressão de CYP1A1, CYP1B1 e CYP2A6 em carcinoma de células escamosas oral

Objective: Tobacco smoke is composed of cancer-causing chemicals referred to as carcinogens. These carcinogens are metabolized by the enzymes of the cytochrome P450 (CYP) family. Our objective was to evaluate the correlation of tobacco consumption parameters with CYP1A1, CYP1B1 and CYP2A6 expression using qRT-PCR in samples of oral squamous cell carcinoma (OSCC). Material and Methods: The sample was divided into 2 groups: Cancer (36 subjects) and non-Cancer (12 subjects). The smokers' participants (36) were evaluated regarding their Nicotine dependence (ND) was assessed by the Fagerström test for cigarette dependence (FTCD). Questions regarding tobacco consumption like the number of cigarettes/day (CPD), duration of use, and pack-years were also evaluated. The Mann-Whitney and Spearman correlation tests were used at a significance level of 5%. Results: 48 participants were included, 32 men (66.7%), 36 smokers (75%) and 27 smokers with OSCC (56.3%). Samples of OSCC expressed more CYP1A1, CYP1B1, and CYP2A6. Especially, the CYP1B1 gene was significantly expressed in OSCC samples, regardless gender or tobacco use. No women expressed CYP2A6, as well as, non-smokers did not express the CYP1A1 and CYP2A6 genes. CYP1A1 gene was higher among men (P = 0.021). Conclusion: Lack of exposure to tobacco may justify the absence of CYP1A1 and CYP2A6 expression in non-smokers. The CYP1B1 gene was significantly expressed in the cancer presence despite gender or tobacco use. The assessment of ND and quantification of tobacco consumption are important instruments in monitoring smokers with benign oral lesions and, especially, in the presence of cancer.(AU)

Objetivo: A fumaça do tabaco é composta de substâncias químicas cancerígenas conhecidas como carcinógenos. Esses carcinógenos são metabolizados pelas enzimas da família do citocromo P450 (CYP). Nosso objetivo foi avaliar a correlação dos parâmetros do consumo de tabaco com a expressão de CYP1A1, CYP1B1 e CYP2A6 por qRT-PCR em amostras de carcinoma de células escamosas bucal (CCEB). Material e Métodos: A amostra foi dividida em 2 grupos: Câncer (36 indivíduos) e sem Câncer (12 indivíduos). Os participantes fumantes (36) foram avaliados quanto à dependência nicotínica (DN) pelo teste de Fagerström para dependência de cigarro (TFDC). Questões relacionadas ao consumo de tabaco como número de cigarros / dia (CPD), tempo de uso e anos-maço também foram avaliadas. Os testes de correlação de Mann-Whitney e Spearman foram utilizados com nível de significância de 5%. Resultados: foram incluídos 48 participantes, 32 homens (66,7%), 36 fumantes (75%) e 27 fumantes com CCEB (56,3%). Amostras de CCEB expressaram mais CYP1A1, CYP1B1 e CYP2A6. Especialmente, o gene CYP1B1 foi significativamente expresso em amostras de CCEB, apesar do sexo ou uso de tabaco. Nenhuma mulher expressou CYP2A6, assim como, não fumantes não expressaram os genes CYP1A1 e CYP2A6. O gene CYP1A1 foi maior entre os homens (P = 0,021). Conclusão: A falta de exposição pode justificar a ausência da expressão dos genes CYP1A1 e CYP2A6 entre não fumantes. O gene CYP1B1 foi significativamente expresso na presença de câncer, independentemente do sexo ou do uso de tabaco. A avaliação da DN e a quantificação do consumo de tabaco são importantes instrumentos no acompanhamento de fumantes com lesões bucais benignas e, principalmente, na presença de câncer (AU)

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.

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.

New insights of CYP1A in endogenous metabolism: a focus on single nucleotide polymorphisms and diseases

Cytochrome P450 1A (CYP1A), one of the major CYP subfamily in humans, not only metabolizes xenobiotics including clinical drugs and pollutants in the environment, but also mediates the biotransformation of important endogenous substances. In particular, some single nucleotide polymorphisms (SNPs) for genes may affect the metabolic ability of endogenous substances, leading to some physiological or pathological changes in humans. This review first summarizes the metabolism of endogenous substances by CYP1A, and then introduces the research progress of SNPs, especially the research related to human diseases. Finally, the relationship between SNPs and diseases is discussed. In addition, potential animal models for gene editing are summarized. In conclusion, CYP1A plays an important role in maintaining the health in the body.

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.

Inhibitory effect of flavonoids from Scutellariae Radix on human cytochrome P450 1A / 中国中药杂志

This study investigated the inhibitory effect of eight natural flavonoids in Chinese herb Scutellariae Radix on huamn cytochrome P450 1 A(CYP1 A), a key cancer chemo-preventive target. In this study, phenacetin was used as a probe substrate for CYP1 A, while human liver microsomes and recombinant human CYP1 A enzymes were used as enzyme sources. Liquid chromatography-tandem mass spectrometry was used to monitor the formation rates of acetaminophen, the O-deethylated metabolite of phenacetin. The dose-dependent inhibition curves were depicted based on the changes of the formation rates of acetaminophen, while the IC_(50) were determined. Inhibition kinetic analyses and docking simulations were used to investigate the inhibition modes and mechanism of wogonin(the most potent CYP1 A inhibitor in this herb), while the inhibition constants(K_i) of wogonin against both CYP1 A1 and CYP1 A2 were determined. Among all tested flavonoids, wogonin, 7-methoxyflavanone and oroxylin A displayed a strong inhibitory effect on CYP1 A(IC_(50)100 μmol·L~(-1)). Further investigations demonstrated that wogonin had a weak inhibitory effect on other human CYP enzymes, suggesting that it could be used as a lead compound for the development of specific inhibitors of CYP1 A. Furthermore, the inhibition kinetic analyses clearly demonstrated that wogonin could strongly inhibit phenacetin O-deethylation in both CYP1 A1 and CYP1 A2 in a competitive manner, with K_i values at 0.118 and 0.262 μmol·L~(-1), respectively. Molecular docking demonstrated that wogonin could strongly interact with CYP1 A1 and CYP1 A2 via hydrophobic and π-π interactions, as well as Ser120 and Ser116 in CYP1 A1 via hydrogen-bonding. In conclusion, this study found that some flavonoids in Scutellariae Radix displayed a strong inhibitory effect on CYP1 A, while wogonin is the most potent CYP1 A inhibitor with a relatively high selectivity towards CYP1 A over other human CYPs.

Effects of CYP1A enzyme specific inhibitor on pharmacokinetics of para-acetaminophen in Bactrian camel

The effects of CYP1A enzyme on the pharmacokinetics of p-acetaminophen were studied in Bactrian camel. Twelve Bactrian camels were divided into 2 groups, then given a single dose of p-acetaminophen only or with the enzyme inhibitor lomefloxacin. Blood samples were collected after different intervals, and p-acetaminophen concentration was determined by high-performance liquid chromatography. Pharmacokinetic parameters were analyzed by Phoenix WinNonLin v.7.0. The results show that lomefloxacin can significantly inhibit Bactrian camel CYP1A enzyme, as evidenced by the prolonged elimination half-life, increased maximum plasma concentration and area under the curve values and the shortened time to peak concentration for p-acetaminophenol in the substrate with inhibitor group. The results lay a foundation for revealing the particular characteristics of the CYP1A enzyme in Bactrian camels.

Association of polycyclic aromatic hydrocarbon exposure and CYP1A1 gene and enzyme activity of peripheral blood monocyte cells in coke oven workers / 中华劳动卫生职业病杂志

Objective@#To explore the association between polycyclic aromatic hydrocarbons (PAHs) exposure and cytochrome P450 CYP1A1 expression at gene and enzyme activity levels in the peripheral blood monocyte cells in coke oven workers, and to provide a certain basis for the biological monitoring of health damage in coke oven workers.@*Methods@#We surveyed 118 coke oven workers and 63 controls (energy power workers in the same company) using self-designed questionnaire, determined their post-shift urinary 1-hydroxypyrene (1-OH-Py) concentration using high performance liquid chromatography (HPLC)-fluorescence detector method. We also isolated the peripheral blood mononuclear cell (PBMC) from fasting venous blood, and detected DNA damage using comet assay, CYP1A1 mRNA level using the real-time fluorescent quantitative PCR (FQ-PCR), and EROD activity using spectrophotometry. Statistical analyses including one-way analysis of variance and multiple linear regressions were used to analyze the association of urinary 1-OH-Py and CYP1A1 mRNA level and EROD activity.@*Results@#Compared to the control group, the urinary 1-OH-Py concentration and PBMC DNA tail moment were significantly increased in coke oven workers (P<0.05), and CYP1A1 gene level and EROD activity in PBMC were significantly decreased (P<0.05). Multiple linear regression showed that a ten-fold increase of urinary 1-OH-Pycon centration was associated with a decrease of 0.77 (95%CI: -1.33--0.21) in CYP1A1 gene level, and a decline of 0.15 (95%CI: -0.76--0.16) in EROD activity of PBMC in coke oven workers (P<0.05).@*Conclusion@#Occupational PAHs exposure induced DNA damage, which was associated with the decreased level in CYP1A1 gene cavel and EROD activity in PBMC of coke oven workers.

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.

Analysis of promoter methylation of CYP1A1 and CYP1B1 gene in thyroid cancer / 中华内分泌代谢杂志

Objective To investigate the promoter methylation of drug metabolism genes CYP1A1 and CYP1B1 in thyroid cancer and its relationship with clinical pathological characteristics. Method 201 cases of thyroid cancer and 23 cases of normal thyroid tissues were involved. Methylation-specific PCR ( MSP ) was performed to analyze promoter methylation of CYP1A1 and CYP1B1 genes in the above tissues to detect the frequency of methylation positive, compare the promoter methylation level of CYP1A1 and CYP1B1 genes in papillary thyroid carcinomas ( PTC) and the controls. Five thyroid cancer cell lines were treated with methyltransferase inhibitor 5-Aza-dC for 5 days, and real time PCR ( RT-PCR) was performed to evaluate the mRNA expression of CYP1A1 and CYP1B1 genes. Logistic regression was used to analyze the correlation between the aberrant methylation and the clinical features. Results Aberrant methylation status in promoter region of CYP1A1 and CYP1B1 genes were detected in all kinds of thyroid cancers. Compared with control tissues, the methylation in promoter regions of CYP1A1 in PTCs was significantly higher, while that in promoter regions of CYP1B1 was lower (P<0.05). In vitro, 5-Aza-dC treatment significantly increased the CYP1A1 gene mRNA expression for 6. 92 and 8. 30 times in K1 and C643 cell lines respectively and restored CYP1B1 gene mRNA expression for 7.62 times in K1 cell line. Compared with the controls, PTCs with methylation in promoter regions of CYP1B1 had decreased lymphatic metastasis rate. Conclusion The methylation in promoter regions of CYP1A1 and CYP1B1 gene may regulate their mRNA expressions. Aberrant methylation of the promoter region of CYP1B1 is associated with lymph node metastasis in PTC.

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.

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.

Study of genetic detection of KRAS, FCGR, CYP3A5 and CYP1A1 in predicting clinical outcomes of chemotherapy/target therapy in metastatic colorectal cancer (mCRC) / 中国医师杂志

Objective To investigate the predictive and prognostic value of genetic detection of Kirsten rat sarcoma viral oncogene homolog(KRAS),Fc gamma receptors (FCGR),cytochrome P450 3A5 (CYP3A5) and CYP1A1 in patients with metastatic colorectal cancer (mCRC) receiving C225 combined with CapeOx chemotherapy.Methods Twelve KRAS wild-type (WT) mCRC patients were selected receiving C225 (cetuximab) combined with CapeOx (capecitabine/oxaliplatin) chemotherapy.KRAS,FCGR,CYP3A5,CYP1 A1 gene mutation were detected before treatment.The expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) were detected in colorectal cancer tissues and the corresponding adjacent tissues expression with immunohistochemical staining (SP method).The relationship between FCGR,CYP3A5,CYP1A1 mutation,and PTEN expression and survival time were analyzed.Results The mutation rates of FCGR,CYP3A5,and CYP1 A1 were 16.7％ (2/12),25％ (3/12) and 16.7％ (2/12) in 12 patients with KRAS WT,respectively.PTEN was expressed mainly in the nucleus in yellowish brown.The rates of expression in the tissue adjacent to the cancer and in the tumor tissue were 100％ (12/12) and 41.7％ (5/12),respectively.PTEN expression in tumor tissue was reduced or absent.The study indicated that the objective response rate [complete response + partial response (CR + PR)] was 80％ in patients whose KRAS,FCGR,CYP3A5,and CYP1A1 were all in wild type,the CR + PR rates in FCGR,CYP3A5,and CYP1A1 gene mutation group were 0,33.6％ and 50％.The objective response rates of patients with PTEN expression or null were 50％ and 37.5％,respectively (P ＜ 0.05).The progression free survival (PFS) with KRAS/FCGR/CYP3A5/CYP1A1 wild type or mutation were 15.56 or 8.12 months (P ＜ 0.05).The overall survival (OS) with wild type or mutation of KRAS/FCGR/CYP3A5/ CYP1A1 were 25.03 or 19.21 months(P ＜0.05).The PFSs of positive or negative expression of PTEN in patients were 9.13 or 7.87 months (P ＜0.05),and the OSs of positive or negative expression of PTEN in patients were 24.25 or 18.74 months (P ＜0.05).Conclusions FCGR,CYP3A5 and CYP1A1 mutations and PTEN expression null both have been associated with resistance to cetuximab in mCRC patients.FCGR,CYP3A5,CYP1A1 and PTEN can be served as the predictive biomarkers for the response to cetuximab.

Ginsenoside Rb1 Inhibits Doxorubicin-Triggered H9C2 Cell Apoptosis via Aryl Hydrocarbon Receptor

Doxorubicin (DOX) is a highly effective chemotherapeutic agent; however, the dose-dependent cardiotoxicity associated with DOX significantly limits its clinical application. In the present study, we investigated whether Rb1 could prevent DOX-induced apoptosis in H9C2 cells via aryl hydrocarbon receptor (AhR). H9C2 cells were treated with various concentrations (−μM) of Rb1. AhR, CYP1A protein and mRNA expression were quantified with Western blot and real-time PCR analyses. We also evaluated the expression levels of caspase-3 to assess the anti-apoptotic effects of Rb1. Our results showed that Rb1 attenuated DOX-induced cardiomyocytes injury and apoptosis and reduced caspase-3 and caspase-8, but not caspase-9 activity in DOX-treated H9C2 cells. Meanwhile, pre-treatment with Rb1 decreased the expression of caspase-3 and PARP in the protein levels, with no effects on cytochrome c, Bax, and Bcl-2 in DOX-stimulated cells. Rb1 markedly decreased the CYP1A1 and CYP1A2 expression induced by DOX. Furthermore, transfection with AhR siRNA or pre-treatment with AhR antagonist CH-223191 significantly inhibited the ability of Rb1 to decrease the induction of CYP1A, as well as caspase-3 protein levels following stimulation with DOX. In conclusion, these findings indicate that AhR plays an important role in the protection of Ginsenoside Rb1 against DOX-triggered apoptosis of H9C2 cells.

Study on the induction of drug metabolizing enzyme CYP1 A1 activity by Ginsenoside Rc,Re,Rf and Rg1 / 中国药理学通报

Aim To investigate the induction effect of ginsenoside Rc, Re, Rf and Rg1 on CYP1A1, and further validate the role of aryl hydrocarbon receptor in CYP1A1 expression. Methods Dual luciferase re-porter gene system was performed. Four kinds of gin-senoside were screened for aryl hydrocarbon receptor activation by reporter assays, and TCDD as the positive control. Further with different concentrations of ginsen-oside Rc, Re, Rf and Rg1 treated on LS174T cells, RNA and total protein were extracted to detect the reg-ulating effect of ginsenosides on CYP1 A1 mRNA and protein expression with Real-time PCR and Western blot technology respectively. Results Reporter gene screening showed that the ginsenoside Rc, Re, Rf and Rg1 could activate AhR and had potential effects on the induction of CYP1A1 enzyme. Meanwhile, dose-de-pendent induction of the gene expression were observed in response to ginsenoside Rc, Re, Rf and Rg1 and the levels of CYP1 A1 protein expression were increased by ginsenoside Rc, Re, Rf and Rg1 in varying de-grees. Conclusion Ginsenoside Rc, Re, Rf and Rg1 can up-regulate the gene and protein expression of CYP1 A1 possibly via the AhR-mediated CYP1 A1 path-way.