科研之友
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摘要
Background: Both total astragalus saponins (AST) and it's main component astragaloside IV (ASIV) have been used in China as cardiovascular protective medicines. However, the anti-inflammatory activities that are beneficial for cardiovascular health have never been compared directly and the molecular mechanisms remain unresolved. This study was conducted to compare the inhibitory effects of these drugs on TNF alpha-induced cell responses, related signaling pathways, and the underlying mechanisms in mouse arterial endothelial cells. Methodology/Principal Findings: Real-time qRT-PCR was performed to determine the expression of cell adhesion molecule (CAM) genes. Immunofluorescent staining was used to detect the nuclear translocation of transcription factor NF-kappa B-p65. Western Blot analysis was used to identify TNF alpha-induced NF-kappa B-p65 phosphorylation, I kappa B alpha degradation, and caspase-3 cleavage. Cell surface proteins were isolated and TNF alpha receptor-1(TNFR1) expression was determined. The results suggest that both AST and ASIV attenuate TNF alpha-induced up-regulation of CAMs mRNA and upstream nuclear translocation and phosphorylation of NF-kappa B-p65. However, TNFR1-mediated I kappa B alpha degradation, cleavage of caspase-3 and apoptosis were inhibited only by AST. These differences in the actions of AST and ASIV could be explained by the presence of other components in AST, such as ASII and ASIII, which also had an inhibitory effect on TNFR1-induced I kappa B alpha degradation. Moreover, AST, but not ASIV, was able to reduce TNFR1 protein level on the cell surface. Furthermore, mechanistic investigation demonstrated that TNFR1-mediated I kappa B alpha degradation was reversed by the use of TAPI-0, an inhibitor of TNF alpha converting enzyme (TACE), suggesting the involvement of TACE in the modulation of surface TNFR1 level by AST. Conclusion: ASIV was not a better inhibitor than AST, at least on the inhibition of TNF alpha-induced inflammatory responses and TNFR1-mediated signaling pathways in AECs. The inhibitory effect of AST was caused by the reduction of cell surface TNFR1 level, and TACE could be involved in this action.
