The Expression Of Hsa-miR-199a-5p In Essential Hypertension And Its Mechanism

Objective:To investigate the expression profile changes of mi-RNAs in peripheral blood of patients with essential hypertension.To investigate the role and the mechanism of hsa-mir-199a-5p in vascular endothelial injury in essential hypertension.Methods:87 patients with essential hypertension and 31 healthy persons were recruited as subjects.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to further verify the expression of hsa-mir-199a-5p in all patients with essential hypertension and healthy.Human umbilical vein endothelial cells(HUVEC)were cultured in vitro and divided into NC group(scramble control microRNA),hsa-microRNA-199a-5p mimics group and Rescue group.Control microRNAs,hsa-microRNAs-199a-5p mimics and hsa-microRNAs-199a-5p mimics + inhibitors were transfected into HUVEC NC group and HUVEC Rescue group respectively.CCK-8,flow cytometry were used to explore the effects of hsa-mir-199a-5p on cell proliferation,cell cycle,cell apoptosis and cell migration of HUVECs.Western Blot Western blot hybridization assay down-regulated the expression of hsa-mir-199a-5p,the target gene AMPK protein of hsa-mir-199a-5p in HUVECs and ULK1 protein in signaling pathway.Results:The expression level of hsa-microRNA-199a-5p in the serum of patients with essential hypertension was significantly higher than that of healthy persons.The results of CCK-8 in vitro showed that the proliferation of vascular endothelial cells(HUVECs)was significantly enhanced after down-regulating the expression of hsa-mir-199a-5p,suggesting that hsa-mir-199a-5p has the ability to inhibit the proliferation of vascular endothelial cells(HUVECs).Flow cytometry showed that the G1/S phase transition of HUVECs in Rescue group was faster than that in NC group,indicating that overexpression of hsa-mir-199a-5p in vitro could regulate the G1/S phase transition of HUVECs and inhibit the proliferation of HUVECs,thereby inhibiting the repair ability of vascular endothelial cells.Flow cytometry results showed that the apoptosis rate of human umbilical vein endothelial cells was significantly reduced after hsa-mir-199a-5p inhibitor transfection(p > 0.05),indicating that in the process of the occurrence and development of primary pupil blood pressure,reducing the expression level of hsa-mir-199a-5p could inhibit the process of HUVECs cell apoptosis.Compared with NC group,the fluorescence values of pre-mimics co-transfected withpre-mimir-199a-5p mimics and pMIR-REPORTTM Luciferace-AMPK-AMPK-WT-UTR luciferase wild-type reporter plasmids significantly decreased after co-transfection(p < 0.05);pre-mimics-199a-199a-5p and pMIR-REPORTTM Luciferace-AMferace-AMPK-MUT-UTR co-transfected with pMIR-REPORTTM Luciferace-AMferace-AMPK-MUT--UTR,no significant difference(p >0.05)(p > 0.Conclusion All right.Western blot immunoblotting showed that the expression of AMPK protein and ULK1 protein in HUVECs increased significantly after down-regulation of has-mir-199a-5p(p < 0.05),suggesting that the biological effect of has-mir-199a-5p may be related to the expression of AMPK-ULK1.CONCLUSION:The expression level of hsa-microRNA-199a-5p in the serum of patients with essential hypertension was significantly higher,it is related to clinical hypertension stage.Over-expression of hsa-mir-199a-5p can inhibit the proliferation of HUVECs through the expression of AMPK-ULK1 signaling pathway,promote the apoptosis of HUVECs,and then promote the injury of vascular endothelium.