我国金融租赁公司风险管控研究.doc
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1、ClofibrateCat. No.: HY-B0287CAS No.: 637-07-0Molecular Formula: CHClOMolecular Weight: 242.7Target: PPARPathway: Cell Cycle/DNA DamageStorage: Pure form -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 monthSolvent PPAR, EC50: 500 MIn Vitro Clofibrate is a PPAR agonist, with E50s of 50 M, null50
2、0 M for murine PPAR and PPAR, and 55 M, null500 M for human PPAR and PPAR, respectively1. Clofibrate (0.5, 1, 2 mM) increases FABP1 expression in two fatty acid (FA)-treated rat hepatoma cells. Clofibrate lowers ROS levels after early treatment, much more than late treatment in FA-treated cells2.In
3、Vivo Clofibrate (0.5%) up-regulates serum concentrations and hepatic expression of FGF21 in fetuses, with a return to basal levels after Clofibrate administration withdrawal. Clofibrate administration-offspring have significantly higher expression of thermogenic genes (Ucp1, Cidea, Ppara Ppargc1a, C
4、pt1b) and UCP1 protein levels in response to HFD in inguinal fat, but not in retroperitoneal (combined with perirenal) or epididymal fat3.Product Data Sheet InhibitorsAgonistsScreening Librarieswww.MedChemE1PROTOCOLCell Assay 1 Cells are seeded at a density of 2.5 104 cells/well (for WST-1, intracel
5、lular lipid droplet quantification and dichlorofluorescein (DCF) assay, 96-well plates) and 1 105 cells/well (for Nile Red Staining, 12-well plates) in MEM/EBSS medium and incubated overnight for adherence. The next day cell culture medium is replaced with freshly prepared medium containing the fatt
6、y acid mixture oleate:palmitate (2:1) in presence of 3% fatty-acid-free bovine serum albumin. Cells are treated with 0, 0.5, 1, 2, and 3 mM fatty acid (FA) mixture for 24 and 48 hr at 37C in a humidified incubator in an atmosphere of 95% air and 5% CO2. Clofibrate is used to increase levels of FABP1
7、 in treated cell cultures. Clofibrate (500 M) is dissolved in DMSOand later added to the medium (DMSO 0.1% v/v in final volume). Control cells are incubated with DMSO alone. Four different cell treatments includ 1-day FA treatment, 2-day FA treatment, early clofibrate intervention and late clofibrat
8、e intervention1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Administration 3Female and male C57BL/6JNarl mice are used for breeding. Females with parity from 1 to 5 are used. Pregnant females are fed either a control (C) or experimental (CF)
9、diet from breeding to parturition. The C diet is based on an AIN-93M diet with a slight modification to contain 21 kcal% fat from soybean oil, whereas the CF diet is the C diet with addition of 0.5% clofibrate. Pregnancy is dated by the presence of a vaginal plug (defined as pregnancy day 1). After
10、spontaneous parturition (pregnancy day 19.5 0.5), all littermates are uniformly nursed by dams fed the C diet for 3 wk, with litter sizes adjusted to 8-10, weaned onto a nonpurified standard diet for 4 wk, and then switched to a HFD (51 kcal% fat, butter-based) for 5 wk. In this study, only male off
11、spring are used and 2 groups of offspring are designated, according to their mothers diet (C or CF). All mice are kept in a room maintained at 23 2C, with a controlled 12-h-light:-dark cycle with ad libitum to feed and drinking water. Body weight and feed intake are recorded weekly3.MCE has not inde
12、pendently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Willson TM, et al. The PPARs: from orphan receptors to drug discovery. J Med Chem. 2000 Feb 24;43(4):527-50.2. Chen Y, et al. Clofibrate Attenuates ROS Production by Lipid Overload in Cultured Rat Hepatoma Ce
13、lls. J Pharm Pharm Sci. 2017;20(0):239-251.3. Chen SH, et al. Prenatal PPAR activation by clofibrate increases subcutaneous fat browning in male C57BL/6J mice fed a high-fat diet during adulthood. PLoS One. 2017 Nov 2;12(11):e0187507.Caution: Product has not been fully validated for medical applErdo
14、steineCat. No.: HY-B0289CAS No.: 84611-23-4Molecular Formula: CHNOSMolecular Weight: 249.31Target: NF-BPathway: NF-BStorage: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 monthSolvent group 2, Erdosteine-treated; group 3, Methotrexate (MTX)-treated; and group 4, Methotrexate+Erdosteine
15、 treated. On the first day of experiment, a single dose of Methotrexate is intraperitoneally administered to groups 3 and 4, although a daily single dose of Erdosteine is orally administered to group 2 and 4 for 7 days. At the end of the experiment, the testes of the animals are removed and weighed.
16、 The levels of total antioxidant capacity and total oxidative stress, and myeloperoxidase activity in the Methotrexate group are higher than the control group (p0.05). Lipid peroxidation levels are not changed in Methotrexate group compared with control group. In conclusion, Erdosteine can effective
17、ly protect the testes in Methotrexate-induced toxicity. Erdosteine administration with Methotrexate improves testicular injures, as indicated by appearance of spermatogenesis in seminiferous tubules2.PROTOCOLCell Assay 1 The murine macrophage/monocyte cell line RAW264.7 are maintained as monolayers
18、in Dulbeccos modified Eagles medium (DMEM) containing 10 % fetal bovine serum, 60 U/mL Penicillin, and 100 g/mL Streptomycin at 37.8C in 5 % CO2. The cell viability is quantified using a colorimetric tetrazolium compound MTS assay. Briefly, 1104 cells incubated with various concentrations of Erdoste
19、ine (1, 10, or 100 g/mL) for 24 h are treated with 10 L of MTS solution (5 mg/mL) for 45 min. The cells are then lysed with isopropyl alcohol, and the absorbance is read at the wavelength of 540 nm1. MCE has not independently confirmed the accuracy of these methods. They are for reference only.Anima
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