The aim of this study was to examine whether flavin-containing monooxygenase (FMO) protein was expressed in cultured rat brain microvascular endothelial cells (BMECs), which constitute the blood-brain barrier (BBB), a...The aim of this study was to examine whether flavin-containing monooxygenase (FMO) protein was expressed in cultured rat brain microvascular endothelial cells (BMECs), which constitute the blood-brain barrier (BBB), and whether N-oxide from the tertiary amine, d-chlorpheniramine, was formed by FMO in rat BMECs. BMECs were isolated and cultured from the brains of three-week-old male Wistar rats. The expression of FMO1, FMO2 and FMO5 proteins was confirmed in rat BMECs by western blotting analysis using polyclonal anti-FMO antibodies, but FMO3 and FMO4 proteins were not found in the rat BBB. Moreover, N-oxide of d-chlorpheniramine was formed in rat BMECs. The intrinsic clearance value for N-oxidation at pH 8.4 was higher than that at pH 7.4. Inhibition of N-oxide formation by methimazole was found to be the best model of competitive inhibition yielding an apparent Ki value of 0.53 μmol/L, suggesting that N-oxidation was catalyzed by FMOs in rat BMECs. Although FMO activity in rat BMECs was lower than that in SD rat normal hepatocytes (rtNHeps), we suggest that rat BMECs enzymes can convert substrates of exogenous origin for detoxification, indicating that BMECs are an important barrier for metabolic products besides hepatic cells.展开更多
Background:Most duck eggs possess a fishy odor,indicating that ducks generally exhibit impaired trimethylamine(TMA)metabolism.TMA accumulation is responsible for this unpleasant odor,and TMA metabolism plays an essen-...Background:Most duck eggs possess a fishy odor,indicating that ducks generally exhibit impaired trimethylamine(TMA)metabolism.TMA accumulation is responsible for this unpleasant odor,and TMA metabolism plays an essen-tial role in trimethylaminuria(TMAU),also known as fish odor syndrome.In this study,we focused on the unusual TMA metabolism mechanism in ducks,and further explored the unclear reasons leading to the debilitating TMA metabolism.Methods:To achieve this,transcriptome,proteome,and metagenome analyses were first integrated based on the constructed duck populations with high and low TMA metabolism abilities.Additionally,further experiments were conducted to validate the hypothesis regarding the limited flavin-containing monooxygenase 3(FMO3)metabolism ability of ducks.Results:The study demonstrated that liver FMO3 and cecal microbes,including Akkermansia and Mucispirillum,par-ticipated in TMA metabolism in ducks.The limited oxidation ability of FMO3 explains the weakening of TMA metabo-lism in ducks.Nevertheless,it decreases lipid deposition and increases antibacterial activity,contributing to its survival and reproduction during the evolutionary adaptation process.Conclusions:This study demonstrated the function of FMO3 and intestinal microbes in regulating TMA metabolism and illustrated the biological significance of FMO3 impairment in ducks.展开更多
文摘The aim of this study was to examine whether flavin-containing monooxygenase (FMO) protein was expressed in cultured rat brain microvascular endothelial cells (BMECs), which constitute the blood-brain barrier (BBB), and whether N-oxide from the tertiary amine, d-chlorpheniramine, was formed by FMO in rat BMECs. BMECs were isolated and cultured from the brains of three-week-old male Wistar rats. The expression of FMO1, FMO2 and FMO5 proteins was confirmed in rat BMECs by western blotting analysis using polyclonal anti-FMO antibodies, but FMO3 and FMO4 proteins were not found in the rat BBB. Moreover, N-oxide of d-chlorpheniramine was formed in rat BMECs. The intrinsic clearance value for N-oxidation at pH 8.4 was higher than that at pH 7.4. Inhibition of N-oxide formation by methimazole was found to be the best model of competitive inhibition yielding an apparent Ki value of 0.53 μmol/L, suggesting that N-oxidation was catalyzed by FMOs in rat BMECs. Although FMO activity in rat BMECs was lower than that in SD rat normal hepatocytes (rtNHeps), we suggest that rat BMECs enzymes can convert substrates of exogenous origin for detoxification, indicating that BMECs are an important barrier for metabolic products besides hepatic cells.
基金supported by the National Natural Science Foundation of China(31672408)the China Agriculture Research Systems(CARS-40)+1 种基金the National Key Research and Development Program of China(2021YFD1200803)the Program for Changjiang Scholars and Innovative Research Team in University(IRT_15R62).
文摘Background:Most duck eggs possess a fishy odor,indicating that ducks generally exhibit impaired trimethylamine(TMA)metabolism.TMA accumulation is responsible for this unpleasant odor,and TMA metabolism plays an essen-tial role in trimethylaminuria(TMAU),also known as fish odor syndrome.In this study,we focused on the unusual TMA metabolism mechanism in ducks,and further explored the unclear reasons leading to the debilitating TMA metabolism.Methods:To achieve this,transcriptome,proteome,and metagenome analyses were first integrated based on the constructed duck populations with high and low TMA metabolism abilities.Additionally,further experiments were conducted to validate the hypothesis regarding the limited flavin-containing monooxygenase 3(FMO3)metabolism ability of ducks.Results:The study demonstrated that liver FMO3 and cecal microbes,including Akkermansia and Mucispirillum,par-ticipated in TMA metabolism in ducks.The limited oxidation ability of FMO3 explains the weakening of TMA metabo-lism in ducks.Nevertheless,it decreases lipid deposition and increases antibacterial activity,contributing to its survival and reproduction during the evolutionary adaptation process.Conclusions:This study demonstrated the function of FMO3 and intestinal microbes in regulating TMA metabolism and illustrated the biological significance of FMO3 impairment in ducks.
