RESUMO
Objective @# To study the effect and mechanism of action of Silibinin on the differentiation of 3T3-F442A preadipocytes in murine.@*Methods @#The effects of 0-400 μmol / L Silibinin on the proliferation of 3T3-F442A adi- pocytes at 24,48 and 72 h were detected by 3-(4,5-dimethylthiazol-2) -2,5-diphenyltetrazolium bromide ( MTT) assay,and the effects of Silibinin on the adipogenesis of 3T3-F442A adipocytes were visualized by Oil Red O stai- ning ; RT-qPCR , Western blot and ELISA assays were used to detect the effects of Silibinin on 3T3-F442A adipo- cyte differentiation-associated transcription factor CCAAT / enhancer binding protein ( C / EBP) α , C / EBP β , per- oxisome proliferator-activated receptor γ cular endothelial growth factor (VEGF) -α and VEGF receptor 2 (VEGFR-2) ,matrix metalloproteinase (MMP) -2 and MMP-9,mitogen-activated protein kinase (MEK) and phosphorylated MEK (p-MEK) ,and extracellular regu- lated protein kinase (ERK) and phosphorylated ERK (p-ERK) expression. (PPARγ) ,adipocyte protein 2 (aP2) ,adipose generation-associated vas@*Results @# MTT assay showed that the cell proliferation rate of 3T3-F442A preadipocytes decreased after 100,200,and 400 μmol /L Silibinin treatment compared with the control group (P<0. 001) ; Oil Red O staining assay showed that the accumulation of red lipid droplets of the cells in the 160 μmol /L Silibinin assay group significantly decreased ; RT-qPCR assay showed that mRNA expression of C/EBPα , C/EBPβ , PPARγ , aP2,VEGF-α , VEGFR-2,MMP-2,and MMP-9 was down-reg- ulated in 3T3-F442A adipocytes treated with 160 μmol /L Silibinin compared with the control group (P<0. 001) ; Western blot assay showed that protein expression of C /EBPα , C /EBPβ , PPARγ and aP2 was down-regulated in 3T3-F442A adipocytes treated with 160 μmol /L Silibinin (P<0. 001) ,and the phosphorylation level of p-MEK/ MEK and p-ERK/ ERK proteins was down-regulated compared with the control group (P <0. 001) ; ELISA assay showed that the protein concentrations of MMP-2 and MMP-9 in the cell supernatant were down-regulated (P < 0. 001) in 3T3-F442A adipocytes treated with 160 μmol /L Silibinin.@*Conclusion @#Silibinin inhibited 3T3-F442A adipocyte differentiation and adipogenesis through inhibition of the MEK/ ERK pathway and matrix metalloproteinase activity.
RESUMO
Objective To evaluate the potential effect of HLF (Hawthorn leave flavonoids, w/w, 80% flavonoids) against thrombus formation, effect of HLF on hypoxia-treated human umbilical vein endothelial cell (HUVECs) was studied. Method The levels of cytotoxicity and NO upon HUVECs were studied by flow cytometry. Moreover, the level of calcium ion in HUVECs was examined through laser scanning confocal microscopy. Result Data from this study showed that HLF at concentrations of 5 μg/ml and 10 μg/ml decreased the cytotoxicity of hypoxia to HUVECs (P<0.05, P<0.01). The intracellular levels of NO and calcium ion were downregulated by HLF at concentrations of 5 μg/ml (P<0.01; P<0.01) and 10 μg/ml (vs control, P<0.01; P<0.01) too. Conclusion Results observed suggest that HLF protect HUVECs from hypoxia partly through its regulative effect on NO and calcium ion levels.
RESUMO
Aim To investigate the protective effect of ginsenoside Rb1 against apoptosis induced by H_2O_2. Methods H_2O_2 was used to build an oxidative stress-induced injury model in neonatal rat cardiomyocytes. After treated with gensenoside Rb1(20, 40, 80 mg?L -1),the apoptosis rate, the content of malondialdehyde (MDA), and the activity of superoxide dimutase (SOD) of the cardiomyocytes were examined. The intracellular calcium indicated by the fluorescence in cells were measured by the laser confocal microscope. Results Compared with the model group, the apoptosis rate and the content of MDA of the cardiomyocytes decreased greatly (P
RESUMO
Aim To examine the effect of Puerarin on calcium overload in human umbilical vascular endothelial cells(HUVECs).Methods The HUVECs were incubated with Puerarin,then were exposed to H_2O_2,ATP and high K~+.The changes of intracellular free calcium([Ca~(2+)]i) in HUVECs were determined using fluorescent Ca~(2+)-indicator fluo-3/AM with confocal laser scanning microscope.Results Puerarin inhibited the elevation of [Ca~(2+)]i induced by H_2O_2,ATP and high K~+,and the inhibition ratio were 72.2%,56.5% and 78.2% respectively.The effect of Puerarin inhibiting the elevation of [Ca~(2+)]i induced by high K~+ was similar to that of verapamil.Conclusion The mechanism of puerarin reducing the overload of calcium in HUVECs may be related with blocking both voltage-dependent calcium channels and the release of calcium pool in endoplasm.