RESUMO
Since neurotrophic factor is easy to degrade and aggregate, it usually has a short half-life in vitro. To overcome this shortage, neurotrophic factor has been combined with the silk fibroin (SF) membrane to realize less degradation, optimal loading efficiency, sustained release, and good adsorption. By optimizing its binding conditions, main parameters were investigated and its optimal loading efficiency was obtained. bFGF was combined to SF membrane by layer by layer (LbL) static adsorption technique. The natural and nontoxic chondroitin sulfate (CS) was used as a crosslinking agent. Optimization was carried out in three aspects: the concentration of bFGF, the concentration of CS, and the reaction time. This experiment provides a better environment for the growth of cells and offers a new kind material of absorbing neurotrophic factor to meet increasing demand for biological materials.
Assuntos
Animais , Ratos , Técnicas de Cultura de Células , Fator 2 de Crescimento de Fibroblastos , Química , Fibroínas , Química , Células PC12RESUMO
This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.
Assuntos
Animais , Ratos , Anacardiaceae , Química , Angiotensina II , Farmacologia , Animais Recém-Nascidos , Células Cultivadas , Colágeno Tipo I , Genética , Metabolismo , Colágeno Tipo III , Genética , Metabolismo , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas , Farmacologia , Fibroblastos , Biologia Celular , Metabolismo , Flavonas , Farmacologia , Frutas , Química , Miocárdio , Biologia Celular , Metabolismo , Plantas Medicinais , Química , RNA Mensageiro , Metabolismo , Ratos Sprague-DawleyRESUMO
<p><b>OBJECTIVE</b>To investigate the effects of emodin on the proliferation of cultured rat vascular smooth muscle cell (VSMC) induced by angiotensin II.</p><p><b>METHOD</b>VSMCs were cultured by explant method. Cell proliferation model was established by stimulation with Ang II. Cell proliferation was measured by MTT assay to observe the effects of emodin (10, 20, 40 and 80 micromol x L(-1)) and N(G)-nitro-L-arginine methyl ester (L-NAME, 100 micromol x L(-1)) on VSMC proliferation induced by Ang II. The expression of PCNA was measured by immunohistochemical staining. Nitric oxide (NO) level was measured by Griess reagent. Nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS) levels were detected by chemical colorimetric method. mRNA expression of iNOS was measured by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULT</b>Emodin at the doses range from 10 to 80 mol x L(-1) inhibited cell proliferation in a dose and time-dependent manner. The inhibitory effects were partly blocked by 100 mol x L(-1) of L-NAME. Emodin markedly decreased the expression of PCNA in VSMC, increased NO, NOS and iNOS levels, and increased iNOS mRNA expression in VSMC.</p><p><b>CONCLUSION</b>Emodin could inhibite VSMCs proliferation induced by Ang II. Inhibiting the expression of PCNA, increasing the NO secretion and upregulating the iNOS gene expression might be associated with the inhibitory effects.</p>
Assuntos
Animais , Masculino , Ratos , Angiotensina II , Farmacologia , Arginina , Farmacologia , Linhagem Celular , Proliferação de Células , Emodina , Farmacologia , Imuno-Histoquímica , Miócitos de Músculo Liso , Biologia Celular , NG-Nitroarginina Metil Éster , Farmacologia , Óxido Nítrico , Metabolismo , Óxido Nítrico Sintase Tipo II , Genética , Antígeno Nuclear de Célula em Proliferação , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Resveratrol (RESV) is a polyphenolic compound existed in native plants such as grape, fleeceflower root, and peanut, etc. The aim of this study was to investigate the effects in vitro of RESV on adenosine diphosphate (ADP)-induced platelet aggregation, platelet membrane-bound fibrinogen (PFig) its mechanism of action. The effects of RESV and phospholipase Cbeta inhibitor (U73122) on ADP-induced healthy human volunteers platelet aggregation, PFig, and the expression of phospho-phospholipase Cbeta3 (P-PLCbeta3) and total-phospholipase Cbeta3 (T-PLCbeta3) were studied with platelet aggregometer, flow cytometry and Western blotting, respectively. Compared with control group, RESV at 25, 50 and 100 micromol x L(-1) inhibited ADP-induced platelet aggregation and PFig in a dose dependent manner, and RESV at 25 micromol x L(-1) obviously reduced expression of P-PLCbeta3 and ratio of P-PLCbeta3 to T-PLCbeta3 in platelet of healthy human volunteers. Furthermore, RESV and U73122 had additive effect in inhibiting platelet aggregation and PFig. All these suggested that RESV inhibited platelet aggregation and PFig induced by ADP partly through decreasing the activity of PLCbeta of platelets, and that RESV had definite effect of antiplatelet and might be developed as a novel antithrombotic agent.