| Part Ⅰ Expression of ZNF295-AS1 and miR-508-5p in peripheral blood of patients with coronary atherosclerosisObjective:To analyze whether the expression of ZNF295-AS1 and miR-508-5p was different in patients with coronary atherosclerosis and those without coronary atherosclerosis.Methods:From April 2018 to September 2018,70 patients with confirmed coronary atherosclerosis and 70 patients without coronary atherosclerosis after examination were selected as the negative control.The expression of ZNF295-AS1 and miR-508-5p was analyzed.Results:1.lncRNAZNF295-AS1 expression was significantly increased in patients with coronary atherosclerosis,the difference was statistically significant,P<0.001;2.In the serum of patients with coronary atherosclerosis,the expression of miR-508-5p was down-regulated,and the difference was statistically significant,P<0.0001;3.Pearson correlation coefficient analysis found that miR-508-5p and lncRNAZNF295-AS1 expression were negatively correlated in the serum of patients with coronary atherosclerosis,and the difference was statistically significant,P<0.0001.4.Smoking,hypertension,diabetes,and ZNF295-AS1 expression are risk factors for patients with coronary atherosclerosis.Conclusion:In the serum of patients with coronary atherosclerosis,the expression of lncRNAZNF295-AS1 is up-regulated and miR-508-5p is downregulated,and the two are negatively correlated.Its significance needs to be further explored in subsequent studies.Part Ⅱ Based on miR-508-5p/ATG7 axis to explore ZNF295AS1 promotes OX-LDL-induced HUVEC autophagyObjective:To analyze the effect of ZNF295AS1 on autophagy in ox-LDL-treated HUVEC by establishing an endothelial cell model(ox-LDLtreated HUVEC),and further investigate whether the high expression of ZNF295AS1 in peripheral blood of patients with coronary The initial stage of AS is related to endothelial damage caused by ox-LDL.Methods:1.Human umbilical vein endothelial cells(HUVECs)were treated with HUVEC treated with different doses of ox-LDL for 24 hours to establish an endothelial injury model.Cell morphology was observed under a microscope,and the expression of ZNF295-AS1 in the cells was detected.MTT measures cell proliferation and WB detects autophagy-related proteins to determine the effect of ox-LDL on HUVEC autophagy;2.Construct ZNF295-AS1 siRNA to silence ZNF295-AS1.After transfecting HUVEC cells,use ox-LDL to treat the cells and detect cell proliferation and expression of autophagy-related proteins to determine whether ZNF295-AS1 treats cells under ox-LDL.Regulation of autophagy;3.Detect miR-508-5p in HUVEC after ox-LDL treatment to determine the expression difference of miR-508-5p in HUVEC treated by ox-LDL;4.On-line software analysis of RegRNA 2.0 found that ZNF295-AS1 has a potential miR-508-5p binding site.Double luciferase experiments verified that miR-508-5p has targeted binding to ZNE295-AS1,and is in the process of transduction.ZNF295-AS1 siRNA-infected HUVEC was used to detect the expression of miR-508-5p,and to detect the expression of autophagy-related proteins in HUVEC treated by ox-LDL after transfection of miR-508-5pmimic in HUVEC.Cell morphology was observed under a microscope to determine miR-508-5p interacts with ZNF295-AS1;5.Analysis of RegRNA 2.0 online software revealed that miR-508-5p has a potential ATG7 binding site.Double luciferase experiments verified binding.Overexpression of ATG7 in miR-508-5pmimic cells detected ox-LDL induction.HUVEC autophagy to determine the interaction of miR-508-5p with ATG7.Results:1.lncRNAZNF295-AS1 was up-regulated in HUVEC treated with ox-LDL,and the difference was statistically significant,P<0.0001;2.The expression of lncRNAZNF295-AS1 in HUVEC cells treated with ox-LDL after transfection was significantly down-regulated,and the difference was statistically significant,P<0.01;3.miR-508-5p was up-regulated in HUVEC cells treated with ox-LDL after transfection.The difference was statistically significant,P<0.01;4.The expression of miR-508-5p was up-regulated in HUVEC after transfection with miR-508-5pmimic,the difference was statistically significant,P<0.001;the expression of ATG7 was basically unchanged in the mimic NC group,and miR-508-5p mimic The expression level in the group decreased,and the difference was statistically significant,P<0.01.5.Interfering with ZNF295-AS1 up-regulated miR-508-5p expression,resulting in down-regulation of ATG7 expression,and alleviated HUVEC autophagy induced by ox-LDL.Conclusion:ZNF295-AS1 can promote autophagy in ox-LDL-treated HUVEC,and it is involved in the regulation of autophagy in ox-LDL-treated HUVEC through the miR-508-5p/ATG7 axis. |
PDF Full Text Request