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1.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 417-424, 2020.
Artículo en Inglés | WPRIM | ID: wpr-827228

RESUMEN

In recent years, biosynthesis of triterpenoid saponins in medicinal plants has been widely studied because of their active ingredients with diverse pharmacological activities. Various oxidosqualene cyclases, cytochrome P450 monooxygenases, uridine diphosphate glucuronosyltransferases, and transcription factors related to triterpenoid saponins biosynthesis have been explored and identified. In the biosynthesis of triterpenoid saponins, the progress of gene mining by omics-based sequencing, gene screening, gene function verification, catalyzing mechanism of key enzymes and gene regulation are summarized and discussed. By the progress of the biosynthesis pathway of triterpenoid saponins, the large-scale production of some triterpenoid saponins and aglycones has been achieved through plant tissue culture, transgenic plants and engineered yeast cells. However, the complex biosynthetic pathway and structural diversity limit the biosynthesis of triterpenoid saponins in different system. Special focus can further be placed on the systematic botany information of medicinal plants obtained from omics large dataset, and triterpenoid saponins produced by synthetic biology strategies, gene mutations and gene editing technology.

2.
Rev. cuba. med. trop ; 65(3): 328-338, jul.-sep. 2013.
Artículo en Español | LILACS, CUMED | ID: lil-692258

RESUMEN

Introducción: las enzimas desintoxicadoras citocromo P450 monooxigenasas (MFO) constituyen uno de los principales mecanismos de resistencia de Aedes aegypti a insecticidas. En Cuba, aunque la presencia de estas enzimas se ha estudiado in vivo mediante el uso de sinergistas, su actividad no se ha determinado cuantitativamente in vitro, elemento que resulta imprescindible para abordar estudios de resistencia metabólica en los insectos. Objetivo: estandarizar un método para detectar la actividad de las citocromo P450 monooxigenasas in vitro, y determinarla entonces, en larvas y adultos de cepas de referencia de Aedes aegypti. Métodos: se utilizaron 3 cepas de laboratorio de Aedes aegypti seleccionadas por 14 o 15 generaciones con temefos, deltametrina o propoxur, respectivamente, y una cepa susceptible a los insecticidas. Resultados: se establecieron las condiciones para los ensayos de actividad enzimática (concentración de proteínas y de sustrato, 0,4 mg/mL y 12 mmol/L, respectivamente; y tiempo de reacción de 10 min). Hubo un incremento significativo de la actividad de las citocromo P450 monooxigenasas en las cepas resistentes, con una mayor frecuencia fenotípica de este carácter en el estadio larva. Conclusiones: las modificaciones a la técnica para la determinación de la actividad enzimática permitieron discriminar entre mosquitos de cepas susceptibles y resistentes en los estadios larva y adulto, por lo que se cuenta con otra herramienta para la detección de la resistencia metabólica en Cuba


Introduction: cytochrome P450 monooxygenase detoxifying enzymes (MFO) are one of the main resistance mechanisms of Aedes aegypti to insecticides. In vivo studies of the presence of these enzymes have been conducted in Cuba with the use of synergists. However, their activity has not been quantitatively determined in vitro, an indispensable step in studies about metabolic resistance in insects. Objective: standardize a method to detect the activity of cytochrome P450 monooxygenase in vitro, and then determine such activity in larvae and adults of Aedes aegypti reference strains. Methods: the study was based on three laboratory strains of Aedes aegypti selected for 14 or 15 generations with temephos, deltamethrin or propoxur, respectively, and a strain susceptible to insecticides. Results: the conditions for enzyme activity assays were established (protein and substrate concentration: 0.4 mg/mL and 12 mmol/L, respectively, and reaction time: 10 min). There was a significant increase in cytochrome P450 monooxygenase activity in resistant strains, with a higher phenotypic frequency in the larval stage. Conclusions: modifications to the technique used for determination of enzymatic activity made it possible to distinguish between mosquitoes from susceptible and resistant strains in larval and adult stages, providing a new tool for the detection of metabolic resistance in Cuba


Asunto(s)
Aedes/inmunología , Resistencia a los Insecticidas/inmunología
3.
Chinese Pharmacological Bulletin ; (12)2003.
Artículo en Chino | WPRIM | ID: wpr-566298

RESUMEN

Cytochrome P450 oxidoreductase (POR) is the only electron donor for all microsome Cytochrome P450 monooxygenases which are phase I metabolizing enzymes responsible for the metabolism of more than 80% drugs used in clinic.Also,POR metabolizes some anti-tumor prodrugs directly.Therefore,the alteration in POR activity caused by the polymorphisms of POR gene will be of great clinical significance.This review summarizes the newest advancement on the effects of POR polymorphisms on drug metabolism.

4.
Korean Journal of Occupational and Environmental Medicine ; : 604-613, 1997.
Artículo en Coreano | WPRIM | ID: wpr-140105

RESUMEN

The effects of styrene on the induction of cytochrome P-450s (P450), (P4501A1/2, P4502B1/2 and P4502El) and activities of other related enzymes were investigated in the male Sprague Dawley rats which were treated with styrene 500 (S1), 1,000 (S2) 1,500 (S3) mg/kg in olive oil intraperitoneally once a day for two days and sacrificed for the preparation of liver microsomes after 24 hrs. 1. The contents of total protein and P450 in the microsomes derived from the styrene treated groups were slightly higher than those from the control group except those from the 53 group (1,500 mg styrene/kg body weight) . The decreases in microsomal protein contents was prominent in the S3 (p<0.05), but the P450 contents was increased significantly in the S2 (p<0.05). 2. The activities of NADPH-P450 and NADH b5 reductase in hepatic microsomes derived from the treated groups were significantly increased in the treated groups(p<0.05). 3. The activities of PROD were also prominently increased with the treatment of styrene except in 53 group, but the activity of EROD was decreased by styrene treatment. The activities of pNPH in the styrene treated groups were higher than that of the control group (p<0.05). 5. Western blotting with monoclonal antibodies against P4502B1/2 isozymes showed the presence of P4502B1/2 in hepatic microsomes from the rats treated with styrene, and the increase in the densities of immunoblots were corelated with the dosages of styrene. The blot densities against P4501A1/2 and P4502El were slightly increased in the styrene treated groups compared with the control group. These results suggested that styrene could primarily induce P4502B1/2 as major and P4501A1/2 and P4502El in minor forms for the metabolism of styrene in rats.


Asunto(s)
Animales , Humanos , Masculino , Ratas , Anticuerpos Monoclonales , Western Blotting , Citocromo P-450 CYP1A1 , Citocromo P-450 CYP2B1 , Sistema Enzimático del Citocromo P-450 , Citocromos , Isoenzimas , Metabolismo , Microsomas , Microsomas Hepáticos , NAD , Olea , Oxidorreductasas , Ratas Sprague-Dawley , Estireno
5.
Korean Journal of Occupational and Environmental Medicine ; : 604-613, 1997.
Artículo en Coreano | WPRIM | ID: wpr-140104

RESUMEN

The effects of styrene on the induction of cytochrome P-450s (P450), (P4501A1/2, P4502B1/2 and P4502El) and activities of other related enzymes were investigated in the male Sprague Dawley rats which were treated with styrene 500 (S1), 1,000 (S2) 1,500 (S3) mg/kg in olive oil intraperitoneally once a day for two days and sacrificed for the preparation of liver microsomes after 24 hrs. 1. The contents of total protein and P450 in the microsomes derived from the styrene treated groups were slightly higher than those from the control group except those from the 53 group (1,500 mg styrene/kg body weight) . The decreases in microsomal protein contents was prominent in the S3 (p<0.05), but the P450 contents was increased significantly in the S2 (p<0.05). 2. The activities of NADPH-P450 and NADH b5 reductase in hepatic microsomes derived from the treated groups were significantly increased in the treated groups(p<0.05). 3. The activities of PROD were also prominently increased with the treatment of styrene except in 53 group, but the activity of EROD was decreased by styrene treatment. The activities of pNPH in the styrene treated groups were higher than that of the control group (p<0.05). 5. Western blotting with monoclonal antibodies against P4502B1/2 isozymes showed the presence of P4502B1/2 in hepatic microsomes from the rats treated with styrene, and the increase in the densities of immunoblots were corelated with the dosages of styrene. The blot densities against P4501A1/2 and P4502El were slightly increased in the styrene treated groups compared with the control group. These results suggested that styrene could primarily induce P4502B1/2 as major and P4501A1/2 and P4502El in minor forms for the metabolism of styrene in rats.


Asunto(s)
Animales , Humanos , Masculino , Ratas , Anticuerpos Monoclonales , Western Blotting , Citocromo P-450 CYP1A1 , Citocromo P-450 CYP2B1 , Sistema Enzimático del Citocromo P-450 , Citocromos , Isoenzimas , Metabolismo , Microsomas , Microsomas Hepáticos , NAD , Olea , Oxidorreductasas , Ratas Sprague-Dawley , Estireno
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