RESUMO
The paper is to report the pharmacokinetic character of a series of chemical compounds in vitro and in vivo. Metabolism stability of a series of chemical compounds was screened by using rat liver microsomes. The samples of different chemical compounds were combined and then simultaneously detected by LC-MS/MS. Compounds y13, y12 and y11 were screened out by microstability assay in vitro. The pharmacokinetics of compounds y11, y12 and y13 was evaluated by using SD rat. The plasma samples were pooled at the same time. The plasma concentrations were determined by LC-MS/MS. The pharmacokinetic character of two compounds y13, y11 was good by screening in vivo, so they were developed for further research. High-throughput screening of drug candidates in vitro and in vivo was effective, to provide information for the chemical structure information and lower the drug development risk.
Assuntos
Animais , Feminino , Masculino , Ratos , Cromatografia Líquida , Métodos , Avaliação Pré-Clínica de Medicamentos , Métodos , Ensaios de Triagem em Larga Escala , Métodos , Microssomos Hepáticos , Metabolismo , Preparações Farmacêuticas , Sangue , Metabolismo , Farmacocinética , Distribuição Aleatória , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização por Electrospray , MétodosRESUMO
<p><b>AIM</b>To investigate the change of NMDAR1 (zeta 1) subunit expression in temple cortex, frontal lobe, hippocampus and cerebellum of three different group rat after 98 dB wide frequency noise exposure.</p><p><b>METHODS</b>Western Blot and RT-PCR technique, combined with auditory brainstem response (ABR) measurement.</p><p><b>RESULTS</b>(1) Expressions of NMDAR1 (zeta 1) subunit in frontal cortex, temple cortex, hippocampus and cerebellum have no difference, but AD model rat is much weaker than the control group. (2) Expression of NMDAR2A (epsilon 1) in temple cortex for physiological saline groups rat have a mostly increase (plus noise), moreover, those are weakest expression in hippocampus. NMDAR1 (zeta 1) subunit in cerebellum have highest expression, moreover, it is weakest in temple cortex. (3) NMDAR1 (zeta1), NMDAR2A (epsilon 1) subunit expression in hippocampus for three groups rat have a down-regulation after adding noise. (4) NMDAR1 (zeta 1), NMDAR2A (epsilon 1) subunit mRNA expression in control group have no remarkable difference in different cortex. (5) Expressions of NMDAR2A (epsilon 1) in frontal temple cortex, hippocampus for AD model rat are less than that of other groups, weakest in cerebellum, weaker in frontal.</p><p><b>CONCLUSION</b>Wide band frequency noise can reduce the expression of NMDAR1 (zeta 1) subunit in hippocampus and cerebellum of AD model rat, however, the way of regulation is not in the mRNA level. Wide band frequency noise can inhibit the expression of NMDAR2A (epsilon 1) in hippocampus, temple cortex of AD model rat, which has been regulated by mRNA level and have cortex area difference.</p>
Assuntos
Animais , Ratos , Encéfalo , Metabolismo , Córtex Cerebral , Metabolismo , Potenciais Evocados Auditivos do Tronco Encefálico , Ácido Glutâmico , Intoxicação , Ruído , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To study the specific expression of the antisense RNA against hepatitis B virus X (HBX) gene in hepatoblastoma cell line and its anti -HBV activity.</p><p><b>METHODS</b>HBX gene (nt.1370-1827) was amplified by PCR, then cloned into EB virus vector pEBAF which contained human alpha-fetoprotein promoter and enhancer. After transfected into 2.2.15 hepatoma cells and ECV304 human endothelial cells by lipofectin, northern blot, ELISA and real-time qualitative PCR were carried out to assay the expression of HBX mRNA, HBV antigens and HBV DNA level, respectively.</p><p><b>RESULTS</b>The HBX antisense RNA expression vector pEBAF-as-HBX which could be expressed specifically in 2.2.15 hepatoblastoma cells was successfully constructed. Both HBV DNA level and the expressions of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) in 2.2.15 hepatoblastoma cells were inhibited by pEBAF-as-HBX. Compared with those in sense control (pEBAF-s-HBX), the inhibitory rates of HBsAg, HBeAg, and HBV DNA were 37.9%, 36.8%, and 25%, respectively.</p><p><b>CONCLUSIONS</b>The pEBAF-as-HBX expression vector may lead to targeted-expression of HBX antisense RNA in hepatoma cells and shows great inhibition effect on HBV.</p>