A Preliminary Study On The Effect And Mechanism Of Ivabradine On The Senescence Of Vascular Endothelial Cells

Objective:To investigate preliminary study on the effect and mechanism of Ivabradine on D-gal-induced vascular endothelial aging damage.Method:Extraction of the original HUVECs and culture to 3-6 generations to test;CCK8 detection of cell viability screening IVA intervention time and concentration,The groups were divided into control group(Control),aging group(D-gal),IVA group(50 μM,100 μM,200 μM).SA-β-Gal staining for cell senescence and the oxidation related indicators ROS、MDA、SOD detection.The difference of Western Blotting detection SIRT1、TGF-β1 and p21 in each group.Result:(1)The results of CCK8 showed that the cell viability of D-gal group was significantly lower than Control group,compared with D-gal group,the endothelial cell viability of IVA group(50 μM,100 μM,200 μM)was increased after 12 hours of pretreatment(P< 0.01).(2)Accroding to the results of SA-β-Gal staining,compared with the Control group,the positive rate of SA-β-Gal was higher in the D-gal group(P<0.05),compared with D-gal group,the positive rate of IVA was decreased(P< 0.05).(3)As a result of the detection of oxidative parameters,the levels of ROS and MDA increased and the level of SOD decreased in the D-gal group compared with the control group(P<0.05).Compared with D-gal group,the levels of ROS and MDA decreased and SOD increased in IVA group.(4)Western-Blotting results suggest: SIRT1 expression was decreased and TGF-β1 and p21 were increased in the D-gal group(P< 0.05);compared with the D-gal group,SIRT1 was increased and TGF-β1 was decreased in the IVA200 μM group(P< 0.05).P21 expression was significantly decreased in the IVA 100 μM and 200μM groups(P< 0.05).Conclusion:.(1)D-gal can decrease cell viability and promote vascular endothelial aging injury by increasing oxidative stress and reducing the expression of anti-aging protein.(2)IVA can increase the vitality of aging endothelial cells,reduce oxidative stress and reduce the positive rate of aging staining,thus improving the injury of aging endothelial cells,so it has a certain protective effect on vascular endothelial cells.(3)IVA increased SIRT1 expression and decreased TGF-β1 and p21 expression in aging endothelium,so the protective mechanism may be related to the activation of SIRT1/TGF-β1 signaling pathway.