Effects Of Short-Chain Acyl-CoA Dehydrogenase On Hypertensive Vascular Remodeling

Objectiveshort-chain acyl-Co A dehydrogenase(SCAD)is a member of acyl-Co A dehydrogenase family,which specifically decomposes the short chain acyl-Co A,is the first speed limiting step of fatty acid beta oxidation and the key enzyme of fatty acid oxidation.We aimed to study the changes of short-chain acyl-Co A dehydrogenase(SCAD)in the hypertension vascular remodeling from animal level and cell level.To explore the effect of swimming training on vascular remodeling and the changes of SCAD.Investigating the expression of SCAD on apoptosis in human umbilical vein endothelial cells(HUVECS)to explore the effect of SCAD on the apoptosis in HUVECS.To investigate the relationship between SCAD and hypertension vascular remodeling.To search the new molecular markers for vascular remodeling of hypertension and find new drug targets for the prevention and treatment of hypertension and its complications.Methods1.Using spontaneously hypertensive rats(SHR)vascular remodeling model to detect systolic pressure,the diameter of the vascular lumen,the tickness of vascular wall and the content of ROS.To observed the effect of blood pressure and aorta by swimming training in rats.2.Using spontaneously hypertensive rats(SHR)vascular remodeling model to observe the expression of SCAD in the aorta of hypertensive rats.To explore the effects of swimming training on hypertension vascular remodeling and the expression of SCAD.To detect SCAD protein expression by Western-blot.To detect the levels of SCAD m RNA by RT-PCR.Detection of SCAD enzyme activity and to observe the change of the ATP content and fatty acid metabolism.3.Culturing the human umbilical vein endothelial cell(HUVEC).The HUVEC treated with tert-butyl hydroperoxide(t BHP)were used as the model of human umbilical vein endothelial cell apoptosis.Cell viability,SCAD enzyme activity,the levels of SCAD m RNA and the expression of SCAD protein were measured.4.Screening the best interference sequences on SCAD.The human umbilical vein endothelial cells were treated with the best interference sequences si RNA to investigate the changes of the SCAD enzyme activity,the levels of SCAD m RNA and the expression of SCAD protein.To investigate the effects of SCAD interference sequences on the content of ROS and free fatty acid and the cell viability and apoptotic rate.5.HUVECS were infected by recombinant adenovirus containing SCAD gene(Ad-SCAD)and treated by t BHP to establish the model of apoptosis.To investigate the changes of the SCAD enzyme activity,the levels of SCAD m RNA and the expression of SCAD protein.To explore the effects of Ad-SCAD on the content of ROS and free fatty acid and the cell viability and apoptotic rate.To observe the anti apoptosis on HUVECS infected by Ad-SCAD.Results1.To compared with Wistar group.The diameter of vascular lumen decreased in SHR group.The tickness of vascular wall,the ratio of vascular wall and the diameter of vascular lumen,the blood pressure and the content of ROS in SHR group were increased significantly.To compared with SHR group.The diameter of vascular lumen increased in SHR swimming group.The tickness of vascular wall,the ratio of vascular wall and the diameter of vascular lumen,the blood pressure and the content of ROS in SHR swimming group were decreased significantly.Hypertension can lead to vascular remodeling.Swimming endurance training can reduce blood pressure in spontaneously hypertensive rats.effectively improve the vascular remodeling phenomenon,which result in oxidative stress induced by reactive oxygen,in the spontaneously hypertensive rats aorta.2.In vascular remodeling model of hypertension,the expression of SCAD at m RNA and protein levels,the enzyme activity of SCAD,the content of ATP and so on were decreased in SHR group.However,the free fatty acid in serum and aorta were increased in SHR group.After 8 weeks of swimming endurance training,the expression of SCAD at m RNA and protein levels,the enzyme activity of SCAD,the content of ATP and so on were increased in SHR group.However,the free fatty acid in serum and aorta were decreased in the SHR swimming group.3.The human umbilical vein endothelial cell apoptosis model results show that: when the t BHP concentration is 50μM and the reaction time was 12 h,the cell survival rate is about 50%,the SCAD protein expression decreased more significantly in human umbilical vein endothelial cell.4.The best SCAD si RNA interference sequence in human umbilical vein endothelial cells is SCAD si RNA679.To compared with control,the expression of SCAD at m RNA and protein levels,the enzyme activity of SCAD were decreased significantly.The free fatty acid and ROS were increased significantly.The cell viability was decreased but the apoptotic rate was increased significantly.The result consistents with the apoptosis model which stimulated by t BHP in human umbilical vein endothelial cells.5.To compare with the group of t BHP,the SCAD enzyme activity was increased and the protein expression and m RNA levels were increased in the group of Ad-SCAD infected.The content of ROS was decreased significantly in the HUVEC infected by Ad-SCAD.The cell viability was increased and the apoptotic rate was decreased significantly in the HUVEC infected by Ad-SCAD.ConclusionIn summary,our study demonstrated that SCAD is related to the vascular remodeling of hypertension and the apoptosis of human umbilical vein endothelial cells.These novel findings demonstrated that SCAD may be as a new target to prevent and treat the vascular remodeling in hypertension.