Effects Of Serum On Human Umbilical Vein Endothelial Cells In Aging

Objective:This study used human umbilical vein endothelial cells(HUVECs) as experimental materials,and used serum of the healthy elderly and gerontal patients with hypertension as damage factors,we established a injured model of HUVECs that reflect the characteristics of healthy elderly and patients with hypertension.HUVECs was cultured in each serum group,we utilized MTT method to detect the growth of HUVECs,and investigated effects of serum on endothelial cells in aging by testing changes in Nitric Oxide(NO)、Endothelin-1(ET-1) and Reactive Oxygen Species(ROS).Methods:The HUVECs was extracted from newborn umbilical cord which is taken from obstetrics in first affiliated hospital of Nanchang University,then we identified endothelial cells and culturing it 3-5 generations to be standby.We collected 20 serum samples from healthy young,the healthy elderly and gerontal patients with hypertension. The experiment was divided into 4 groups:blank control group(only add culture medium without serum),healthy young group,the healthy elderly group and gerontal hypertension group.We cultured HUVECs in each serum group,and then collected cell supernatants and cell at different times.we utilized MTT method to research the growth of HUVECs,redox method for detecting NO,ELISA method for detecting ET-1,Flow cytometry for detecting intracellular ROS.At last,we uesd statistical methods to process this experimental data,study the differences in each group.Results:1. The cell proliferation of each treatment group at the different time point has no significant difference on statistics(P>0.05),and at the same time point,the cell proliferation of healthy young group,the healthy elderly group and gerontal hypertension group has no significant difference on statistics(P>0.05),but blankcontrolgrouphasdifferentwithothergroups(p<0.05);2.thecontentsofnoinhealthyyounggroup,thehealthyelderlygroupandgerontalhypertensiongroupwasgradientdecreaseafterhuvecswasculturedineachserumgroupfor24hand48h(p<0.01).andtheamountofnoat24hwasmorethan48 h,butithasnosignificantdifferenceonstatistics(p>0.05).3.thecontentsofet-1inhealthyyounggroup,thehealthyelderlygroupandgerontalhypertensiongroupwasgradientincreaseafterhuvecswasculturedineachserumgroupfor24hand48h(p<0.01).andtheamountofet-1at24hwasmore than 48h(P<0.01).4. After HUVECs was cultured in each serum group for 24 h and 48 h,the contents of ROS in healthy young group,the healthy elderly group and gerontal hypertension group show a increasing trend.Conclusions:Serum of the healthy elderly and gerontal patients with hypertension would reduce NO,promote ET-1 and ROS production,but have no significant influence on the proliferation of vascular endothelial cells.