ABSTRACT
Objective To study the metabolic transformation pathways of 4F-MDMB-BUTINACA in vivo by establishing zebrafish models. Methods Six adult zebrafish were randomly divided into blank control group and experimental group, with three fish in each group. After the zebrafish in the experimental group were exposed to 1 μg/mL 4F-MDMB-BUTINACA for 24 h, they were transferred to clean water and cleaned three times, then pretreated for instrumental analysis. The zebrafish in blank control group were not exposed to 4F-MDMB-BUTINACA. Mass spectrometry and structural analysis of 4F-MDMB-BUTINACA and its metabolites were conducted by liquid chromatography-high resolution mass spectrometry and Mass Frontier software. Results A total of twenty-six metabolites of 4F-MDMB-BUTINACA were identified in zebrafish, including eighteen phase Ⅰ metabolites and eight phase Ⅱ metabolites. The main metabolic pathways of phase Ⅰ metabolites of 4F-MDMB-BUTINACA in zebrafish were ester hydrolysis, N-dealkylation, oxidative defluorination and hydroxylation, while the main metabolic pathway of phase Ⅱ metabolites was glucuronidation. Conclusion Metabolite Md24 （ester hydrolysis） and Md25 （ester hydrolysis combined with dehydrogenation） would be recommended to be potentially good biomarkers for abuse of 4F-MDMB-BUTINACA.
Subject(s)
Animals , Cannabinoids , Chromatography, Liquid , Illicit Drugs , Microsomes, Liver/chemistry , ZebrafishABSTRACT
Synthetic cannabinoids are currently a class of new psychoactive substances with the largest variety and most abused. Metabolite identification research can provide basic data for monitoring synthetic cannabinoids abuse, which is the current research hotspot. The main trend of structural modification of synthetic cannabinoid is to replace the fluorine atom on pentyl indole or indazole cyclopentyl with hydrogen atom, which greatly improves the biological activity of the compound. The main metabolic reactions include hydroxylation, fluoropentyl oxidative, ester hydrolyze, amide hydrolysis. Liquid chromatography-high resolution mass spectrometry has become the preferred choice for the structural identification of metabolites. This review mainly summarizes research on metabolism software prediction and human hepatocyte model, human liver microsomes model, rat in vivo model, zebrafish model and fungus C. elegans model in metabolite identification based on the structure and classification of synthetic cannabinoids.
Subject(s)
Animals , Rats , Caenorhabditis elegans , Cannabinoids , Chromatography, Liquid , Microsomes, Liver/chemistry , ZebrafishABSTRACT
El presente trabajo tuvo por objeto estudiar el origen de los ácidos grasos poliinsaturados de cadena larga (AGPI-CL) necesarios para el desarrollo postnatal del cerebro. Ratas normales recién nacidas de 1,5,10 y 20 días de edad fueron decapitadas, inmediatamente después los hígados fueron procesados para obtener la fracción microsomal y determinar la compossición de ácidos grasos y la actividad de la enzima delta6 desaturasa. También se nalizaron: la composición de ácidos grasos en plasma, homogenizado de hemisferios cerebrales y en la leche tomada del estómago de las ratas recién nacidas. Nosotros encontramos que la actividad de la enzima delta6 desaturada en el hígado fue muy baja en comparación a la observada en ratas adultas. Durante el desarrollo postnatal se observaron cantidades apreciables de DHA y AA en la lehe, microsomas hepáticos, lípidos de plasma y hemisferios cerebrales. En conclusión proponemos que durante el desarrollo postnatal, la síntesis hepática contribuye poco en el aporte de DHA y AA para el crecimiento del cerebro, pero tiene la capacidad de concentrar estos ácidos grasos provenientes de la leche, en los microsomas hepáticos y secretarlos hacia el plasma para finalmente llegar al cerebro.
Subject(s)
Animals , Rats , Male , Female , Brain/growth & development , Fatty Acids, Essential/metabolism , Liver/metabolism , Arachidonic Acids/analysis , Arachidonic Acids/metabolism , Docosahexaenoic Acids/analysis , Docosahexaenoic Acids/metabolism , Fatty Acid Desaturases/metabolism , Fatty Acids/analysis , Liver/enzymology , Liver/physiology , Microsomes, Liver/chemistry , Milk/chemistry , Phospholipids/chemistry , Rats, Sprague-DawleyABSTRACT
Human and experimental diabetes mellitus extensively alters lipid metabolism. The eSS is a rat strain that develops a spontaneous diabetes of slow evolution, resembling the non-insulin-dependent diabetes mellitus of young people. We report here disturbances in lipid metabolism of 5-month old eSS rats compared to age-matched alpha-controls. Normal plasmatic glucose levels were found in the fasted state, whereas a diabetic curve was evident for eSS rats after glucose load. Triglyceride content was elevated in plasma and in liver microsomal preparations of eSS animals, when compared to the controls. The diabetic strain revealed a significant fall in the amount of linoleic acid in liver and kidney microsomes and in erythrocyte membranes. In liver, an increase in 22:6 (n-3) was also noted. A depression in the content of linoleic acid as well as an enhancement of docosahexaenoic acid were detected in phosphatidylcholine and phosphatidylethanolamine fractions from liver microsomes of eSS rats. The fatty acid pattern of eSS rat testis showed a raise in the relative percentage of arachidonic and a decrease in 22:5 (n-6), 22:5 (n-3) and 22:6 (n-3) acids compared to their controls. Diabetic rats exhibited a significant increase in microsomal cholesterol content and cholesterol/phospholipid ratio in liver and testis. In the latter tissue, higher values of fluorescence anisotropy were also observed. The current observations indicate that in early stages of the diabetes onset, when eSS rats are still normoglycemic, severe alterations of lipid metabolism may contribute to the establishment and progression of the diabetic syndrome.
Subject(s)
Animals , Rats , Diabetes Mellitus, Type 2/metabolism , Glucose Tolerance Test , Lipids/metabolism , Triglycerides/blood , Cholesterol/analysis , Diabetes Mellitus, Type 2/physiopathology , Disease Models, Animal , Erythrocyte Membrane/chemistry , Fatty Acids/analysis , Kidney/chemistry , Kidney/cytology , Linoleic Acid/analysis , Liver/chemistry , Liver/cytology , Microsomes, Liver/chemistry , Testis/chemistry , Testis/cytologyABSTRACT
El efecto del envejecimiento sobre la composición fosfolipídica de la membrana hepática fue estudiada en dos grupos de ratas Wistar jóvenes (2 meses) y maduros (6 meses). Cuando fue analizado el contenido fosfolipídico microsomal total, el grupo de ratas maduras mostró un incremento significativo (73%). El patrón fosfolipídico microsomal también mostró un comportamiento diferente en ambos grupos, con un significativo incremento en fosfatidilcolina (62%), fosfatidilserina (124%), fosfatidilinositol (31%) y esfingomielina (10%) y aparente en fosfatidiletanolemina y fosfatidilglicerol en el grupo de 6 meses. Una elevada fluidificación de la membrana microsomal en animales añosos fue relevado por un incremento en el índice PC/EM (47%). Este incremento en la fluidificación durante el proceso de envejecimiento puede reflejar una respuesta adaptativa resultante en cambio en la actividad enzimática responsable del metabolismo de drogas y carcinógenos
Subject(s)
Animals , Male , Rats , Aging/physiology , Membrane Fluidity/physiology , Microsomes, Liver/physiology , Phospholipids/chemistry , Body Weight , Microsomes, Liver/chemistry , Organ Size , Rats, Inbred StrainsABSTRACT
El objetivo del presente estudio es determinar el efecto de la interrupción del flujo biliar sobre la detoxicación hepática del lorazepan y su relación con la composición fosfolipídica de la membrana microsomal. Se observó una disminución del contenido microsomal de fosfatidilcolina, fosfatidilinositol, fosfatidiletanolamina y esfingomielina, y un incremento de fosfatidilserina y fosfatidilglicerol. Las ratas control y colestáticas presentaron una actividad enzimática similar para detoxicar el lorazepan. Estos resultados sugieren que la regulación de esta vía metabólica se ejerce con independencia del entorno fosfolipídico de membrana
Subject(s)
Rats , Male , Animals , Bile/physiology , Cholestasis, Extrahepatic/physiopathology , Lorazepam/metabolism , Microsomes, Liver/chemistry , Glucuronosyltransferase/metabolism , Phospholipids/metabolismABSTRACT
The mechanism of glucose transported (GT) expression on the plasma membranes of hepatoma cells in rats induced by 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) was studied. Cytochalasin B binding to plasma membrane fractions from control and 3'-MeDAB group in the absence of cold cytochalasin B showed 9,825 +/- 925 and 30,165 +/- 625 dpm/mg membrane protein. Scatchard plot analysis showed that the GTs present on the plasma membrane fractions in control and 3'-Me DAB groups were 5.0 and 16.0 pmol/mg membrane protein and their Kd values were 151 and 157 nM, respectively. These results suggest that the numbers of GTs in plasma membrane were increased in the 3'-Me DAB group compared to the control group. In contrast, the amounts of GTs in low density microsomal (LDM) fractions measured by a photoaffinity labeling technique using [3H]-cytochalasin B were 31,207 and 11,702 dpm/mg protein in the control and 3'-Me DAB group, respectively. These results suggest that GTs were translocated from LDM to plasma membranes during carcinogenesis. To confirm these results by an independent method 10% SDS-polyacrylamide gel electrophoresis was carried out. Gel slice No. 13 corresponding to MW of 45 kDa from plasma membrane fractions showed increased radioactivities in the 3'-Me DAB group compared to the control group. However, LDM fractions of the 3'-Me DAB group showed decreased radioactivities compared to the control group. Western blot analysis using anti-human RBC GT antibody present in the plasma membranes and LDM fractions from control and 3'-Me DAB groups did not show any significant difference, indicating low cross-reactivity between them. These results indicate that increased glucose transport seems to be more likely due to reciprocal redistribution of GTs between plasma membrane and LDM fractions.