| Background:Endothelial progenitor cells(EPC)is the precursor cells of vascular endothelial cells which plays a key role in the physiological and the pathological angiogenesis and be able to participating in reendothelialization of blood vessel and promoting blood vessel repair.Many experiments have confirmed that cardiovascular risk factors can impaire EPC’s function,and the number of circulating endothelial progenitor cells significantly decreased in patients with cardiovascular risk factors.As an independent risk factor of cardiovascular risk factors,hypertension can damage the endothelial cell function and promote the development of atherosclerosis.In this study we found that the percentage of early and late apoptotic of peripheral blood progenitor cells isolating from patients with hypertension increase compared with the health population and hypertension also enhance the oxidative stress.In addition,hypertension can also damage the function of cells proliferation,migration and adhesion.However,under the stimulation of benazepril,an antihypertensive drug,the percentage of apoptotic cells and oxidative stress of EPC of hypertension patients significantly decreased and cell proliferation,migration,adhesion function significantly improved,indicating that benazepril improved the function of endothelial progenitor cells under the condition of hypertension in a concentration and time dependent manner.In this study,using the SDF-1/CXCR4 axis inhibitors of AMD3100 combined with benazepril treat the EPCs of patient with hypertension,we found that the improment function of benazepril to hyperpietic endothelial progenitor cells was inhibited by the inhibitor of SDF-1/CXCR4,showing that as a traditional cardiovascular risk factors,hypertension seriously damage the function of endothelial progenitor cells.Further research has also revealed that the antihypertensive drugs,benazepril can improve the function of endothelial progenitor cells under the conditions of high blood pressure through the axis of SDF-1/CXCR4.This study is a new exploration of benazepril to treatment of hypertension and provide the theoretical basis for endothelial stem cells to treat hypertension patients.Objective:This study aimed to explore the mechanism of benazeprilimproving the EPC function and oxidative stress in patients with high blood pressure through research the effects of benazepril on the fuctions and the reaction of oxidative stress of endothelial progenitor cells isolated from peripheral blood of patients with hypertension.Content:1.Isolate and culture the EPCs from human peripheral blood,and identify these cells through detecting surface antigen and related indexes.2.Isolate the peripheral blood EPC from healthy group,hypertension group and benazeprilintervented hypertension group,and detectdthe cell proliferation,migration,adhesion,apoptosis and oxidative stress response;3.Detectethe mRNA and proteinexpression of SDF-1 and CXCR4 of EPCs in different groups.Methods:According to "China Hypertension Prevention Guide 2010 Edition" standard selected confirmed 15 patients with grade 1 hypertension.At the same time,15 healthy subjects was included in this study as control group.20 mL fresh anticoagulant of peripheral blood was taken from the vein and then EPC was separated by density gradient centrifugation and cultured in EGM-2 medium,the cell morphology was taken photos.CD133 and CD34 were used to indentify the markers of EPC by flow cytometry after cultured 5 days.Binding experiments further identificated the function of EPC through uptaking Dil-ac-LDL and UEA-1;The early and late apoptotic of EPCs of different groups were detected by Annexin V-FITC/PI.ELISA was used to detect the oxidative stress(MDA and SOD)and SDF-1;The cell proliferation was detected by MTT;The migration ability of EPC cells was detected by Transwell;The adhesion ability of EPC cell was detected in the presence of fibronectin in different groups and different processing conditions through the method of cell counting;Real-time PCR was used to detect the relative mRNA expression of CXCR4 of EPC in different groups and different treatments.Results:1.Cell morphological observation showed that similar patterns of cell of EPC and CD133 and CD34 match the specific surface markers of EPC.At the same time,double positive identification showed that the EPCs have the function of uptaking acLDL-DiI and UEA-1.2.The percentage of apoptosis of early and late apoptotic rate was significantly higher in hypertensive patients compared to healthy subjects.And the intervention of benazepril significantly decreased the apoptosis rate of hyperpietic endothelial progenitor cells.With the increased concentration and prolonged time of benazepril intervention,apoptosis rate gradually decreased in a concentration and time dependent manner.The level of SOD in patients with hypertension significantly decreased,while MDA is reverse.Treatment with benazepril can increase the secretion SOD of EPC of hypertensive patients,and reduce the production of MDA.3.Compared to the control group,proliferation,migration and adhesion ability of EPCs were significantly decreased in patients with hypertension.Under the treatment of benazepri,these functions of EPCs were markedly improvedin a concentration and time dependent manner.4.Compared to the control group,benazepril can significantly decrease the rate of cells apoptosis and oxidative stress response and enhanced the ability of cell proliferation,migration and adhesion.However,SDF-1/CXCR4 inhibitor,AMD3100 significantly abolished the beneficial effect of benazepril on the EPCs from hypertensive patients.The CXCR4 mRNA expression of EPCs decreased in patients with hypertension.Treatment with benazepril can increased the expression of CXCR4.However,AMD3100 abolised the increased expression of CXCR4.In addition,the SDF-1 secretion of EPCs significantly decreased in the patients with hypertension,while benazepril can increase the secretion of SDF-1.However,AMD3100 abolised the increased the secretion of CXCR4.Conclusion:1.EPCs were successfully isolated from hypertensive patients and healthy groups and had the stable morphological after culturing.2.In hypertension patients,EPCs have a higher rate of apoptosis and enhanced oxidative stress,while have a lower ability of proliferation,migration and adhesion.3.Benazepril,an antihypertensive drug,can significantly decrease the rate of cells apoptosis and oxidative stress,promote the proliferation,migration and adhesion ability of EPCs in patients with hypertension in a concentration and time dependentmanner.4.SDF-1/CXCR4 axis plays an important role in improving the function of EPCs in patients with hypertension. |
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