| Objective Through the establishment of arecoline in vitro metabolism system, compare arecoline metabolic ability of SD rats' different organs and liver components.In order to explore arecoline main metabolic enzymes distribution and metabolic features. And by adding inhibitor and components such as purifying method to determine metabolic enzymes.To further research arecoline toxicity mechanism and metabolism to provide the evidence toxicologically significant.Metheds Preparing organ homogenate, collecting blood; Differential centrifugation separating liver subcellular fractions; Etc density gradient centrifugation purificating mitochondria; Establish reaction system to detect arecoline by using ion RP-HPLC. Combined with enzyme inhibitors to compare the metabolic ability of various organs, components and mitochondria before and after purifying.Results 1. Content of arecoline had good linear relationship with the peak detected by Ion RP-HPLC.2. Arecoline metabolic enzymes in SD rats was widespread, especially in the liver and blood. Arecoline metabolic enzymes distributed in the subcellular fractions of SD rat liver cells subcellular fractions.3. Characteristics of arecoline in vitro metabolic accorded with enzymatic reaction;The metabolism enzyme which can etabolize arecoline mainly distributed in the lysosome, mitochondria and microsomal.4. The activity of carboxylic acid ester enzyme was negatively related to the TPP dose. Trace TPP (0.5 nmol/ml) can block the carboxylic acid ester enzyme arecoline metabolism.Conclusion 1. The precision and acurracy of Ion RP HPLC method to detect arecoline was required.2. Arecoline in vitro metabolic belongs to enzymatic reaction.Although the activity of enzymes is different in different organs and components, the vigor of metabolic enzymes basically the same.3. The carboxylic acid ester enzyme may be the main metabolic enzymes of arecoline.4. Arecoline metabolism could be blocked by trace TPP. TPP can be used as tool to reserch arecoline. |
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