Preparation and Characterization of Rodent Intestinal Microsomes: Comparative Assessment of Two Methods
نویسندگان
چکیده
Small intestine plays an important role in the first-pass metabolism of orally ingested xenobiotics as a result of expression of both Phase I and Phase II metabolic enzymes, together with associated transporters. Intestinal microsomes thus can be used to study susceptibility of compounds to metabolism in vitro. The present study was undertaken to have a comparative assessment between different methods of preparation of rodent intestinal microsomes. Mouse and rat intestinal microsomes were prepared by two methods, in method A intestines were homogenized, while in method B mucosal cells were scrapped followed by homogenization. Further, microsomes were prepared by centrifugation (10000xg) followed by ultra centrifugation (100000xg) of the homogenates. The prepared microsomes were characterized for protein concentration using Bradford's method and CYP450 content using carbon monoxide bubbling method. The protein concentration and CYP450 content in microsomes prepared by method B was significantly higher than method A. In conclusion, superior quality intestinal microsomes can be obtained from rodents by using scrapped intestinal mucosal cells as compared to the intestinal homogenates.
منابع مشابه
Quantifying gut wall metabolism: methodology matters
Oral administration continues to be the dominant route for the dosing of small molecules. Therefore having adequate oral bioavailability remains a key component for the success of drug candidates. Amongst various factors determining the overall bioavailability, the role of the intestinal metabolism is commonly overlooked [1]. Intestinal microsomes are commercially available, analogous to hepati...
متن کاملOptimization of intestinal microsomal preparation in the rat: A systematic approach to assess the influence of various methodologies on metabolic activity and scaling factors
The metabolic capacity of the intestine and its importance as the initial barrier to systemic exposure can lead to underestimation of first-pass, and thus overestimation of oral bioavailability. However, the in vitro tools informing estimates of in vivo intestinal metabolism are limited by the complexity of the in vitro matrix preparation and uncertainty with the scaling factors for in vitro to...
متن کاملStudy of zoonotic intestinal helmiths of alimentary tract of rodents in Hamadan, 2012
Abstract Background and Objective: Assessment of alimentary tract helminthes of rodents has a great zoonotic importance. This study aimed at determining the prevalence of helminth infections in rodents in Hamadan. Material and Methods: a total of 100 rodents trapped from different parts of city were transported to laboratory. After anesthetizing by chloroform, the animals ...
متن کاملThe design and preparation of fluorescent labeled chitosan nanoparticles for intestinal delivery
Background: Chitosan is a biodegradable, biocompatible polymer regarded as safe for human dietary. Nanoparticles (NP) prepared with chitosan derivatives typically possess a positive surface charge and mucoadhesive properties such that can adhere to mucus membranes and release the drug payload in a sustained release manner. Current study is focused on preparation several kinds of chitosan formul...
متن کاملI-59: Diagnostic Accuracy of Physical Examination,Transvaginal Sonography, RectalEndoscopic Sonography, and Magnetic ResonanceImaging to Diagnose Deep InfiltratingEndometriosis
Background: To compare the value of physical examination, transvaginal sonography (TVS), rectal endoscopic sonography (RES), and magnetic resonance imaging (MRI) for the assessment of different locations of deep infiltrating endometriosis (DIE). Materials and Methods: Ninety-two consecutive patients with clinical evidence of pelvic endometriosis. Physical examination, TVS, RES, and MRI, perform...
متن کامل