| Objective Observing the protective effect and mechanism of Tanshinone IIA on HUVECs via Molecular biology,cell biology,proteomics and construction of protein-protein interaction networks.These results give a powerful proof for clinical application,and offer an experiment flat for the multi-targets of Chinese Drug.Methods MTT assay was used to determine the optimal concentration of Tanshinone IIA and ox-LDL for Human umbilical vein endothelial cells in vitro.Then cells were treated with different concentrations of Tanshinone IIA toghter with ox-LDL for 24h.?The content of ROS in HUVECs was detected by flow cytometry.?The change of MDA content and SOD activity were detected.?Differential expression proteins were screened out via 2D-DIGE combined with mass spectrometric identification technology.?Use BioPUBINFO website to establish the protein-protein interaction networks in our differential expression proteins.Results ?0-8 mmol/L concentration of TSN?A had no effect on HUVECs proliferation(P>0.05),more than 50 mg/L concentration of ox-LDL had a significant anti-proliferation compared with the normal group(P<0.05).?After flow cytometry tested,the content of ROS increased in HUVECs by ox-LDL(P<0.01),then the content had a gradually drops after TSN?A induced.?TSN ?A+ox-LDL intervention groups induced the content of MDA and increased the activity of SOD compared with ox-LDL(P<0.05).?29 differential expression proteins had been identified via mass spectrometry.We selected 11 critical nodes through analysis of protein-protein interaction networks,and mainly involving pathway were found that cell metabolism,oxidative phosphorylation,cellular components and cytoskeleton.Conclusion?50mg/L ox-LDL treated for 24h could establish HUVECs damage model.?The ability of protecting HUVECs from oxidative injury was enhanced obviously at different concentration TSN IIA and the content of MDA and ROS were decreased while the activity of SOD was increased.?TSN IIA has major regulation through oxidative phosphorylation pathway and anti-inflammatory and antioxidant and exert protective effect on HUVECs. |
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