ABSTRACT
AIM:To explore the influences of semaphorin 3A (Sema 3A) on hydrogen peroxide (H2O2)-induced injury in human umbilical vein endothelial cells (HUVECs).METHODS:Sema 3A over-expression vectors were constructed and transfected into the HUVECs by Lipofectamine 2000, and the over-expression effect was verified by qPCR and Western blot.The HUVECs in different groups were treated with or without 200 μmol/L H2O2 for 4 h.The levels of inflammatory cytokines were measured by qPCR.The levels of lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by corresponding colorimetry.The cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.The levels of apoptosis-related proteins cleaved caspase-3 and Bcl-2 were determined by Western blot.RESULTS:H2O2 induced inflammatory cytokine secretion, increased the levels of LDH and MDA, decreased SOD activity and cell viability, and increased cell apoptosis in the HUVECs.Over-expression of Sema 3A enhanced the above processes.No injury effect of Sema 3A over-expression on HUVECs without H2O2 treatment was observed, indicating that the injury effects of Sema 3A on HUVECs depended on H2O2.CONCLUSION:Sema 3A markedly enhances H2O2-induced injury in the HUVECs, which depends on H2O2.Sema 3A may promote oxidative stress-caused endothelial cell injury.
ABSTRACT
Aim To investigate the role of Wnt/β-cate-nin signaling pathway on the baicalin-induced osteo-genic differentiation in rat bone marrow derived mesen-chymal stem cells ( rBMSC ) . Methods rBMSC was isolated and cultured by adherence screening method. Alkaline phosphatase ( ALP) amount, CFU-FALP and mineralized nodules were compared between each ba-icalin group and vehicle control group at different time points. Real time q-PCR was employed to evaluate the mRNA level of Wnt signaling-related marker ( Wnt10a, GSK-3β,β-catenin and LEF1) after baica-lin treatment. Protein expression of β-catenin and Runx2 was measured by Western blot. Results Ba-icalin significantly increased ALP activities from day 3 to day 7 . The formation of CFU-FALP and mineralized nodules remarkably increased after rBMSC was treated with1, 10, 50 μmol · L-1 baicalin. mRNA levels of Wnt10a, β-catenin, GSK-3β, LEF1and osteocalcin were enhanced significantly in baicalin-treated group compared to control group. Protein expression of β-catenin and Runx2 was also elevated. Conclusion Baicalin ( 0. 1 to 50 μmol · L-1 ) promotes the osteo-genic differentiation and maturation of rBMSC, in which Wnt/β-catenin signaling pathway might be in-volved.
ABSTRACT
To explore novel antifatigue agents targeting with AMPA receptor, 10 compounds were synthesized and their structures were confirmed by 1H NMR, ESI-MS and elemental analysis. 1-BCP was treated as the leading compound. The antifatigue activities were evaluated by weight-loaded forced swimming test, and the AMPA receptor binding affinities were tested with radioligand receptor binding assays. The results unveiled that 5b appeared to possess potent antifatigue activities and high affinity with AMPA receptor, which deserved further studies.