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Differential expression profile and bioinformatics analysis of lncRNAs chip in ApoE-/-mouse atherosclerosis modelObjective:In order to find lncRNA that can be used for clinical diagnosis,an atherosclerosis model was constructed to find the differential expression profile of lncRNAs and mRNAs,the functions and pathways involved in differentially expressed lncRNAs were analyzed,and interested lncRNAs were further studied.Methods:After being fed with high-fat diet for 8 weeks,the ApoE knockout mice were tested for blood lipid levels,and then the whole aorta was stained with oil red O to construct an atherosclerosis model.Group sequencing analysis,GO and Pathway analysis of differential lncRNA associated mRNAs.Results:1.After 8 weeks of treatment,the TC,HDL-C,LDL-C blood lipid levels of mice in the high-fat diet group were significantly higher than those in the normal diet group(P<0.01);the oil red O staining in the experimental group showed red artery The positive rate of atherosclerotic plaque was as high as 68%,which was significantly increased compared with the control group(15%).2.Compared with the blood vessels in the control group,a total of 4711 differentially expressed lncRNAs were detected in atherosclerotic blood vessels,of which 2209 were up-regulated,2502 were down-regulated,another 2191 mRNA were up-regulated,and 1302 mRNA were down-regulated(Fold change? 2.0).3.GO analysis showed that mRNAs related to lncRNAs among differentially expressed genes mainly involved biological functions such as DNA methylation,positive epigenetic regulation,nucleosome assembly,tricarboxylic acid cycle,protein binding,Poly(A)tail,etc.;Pathway analysis of the KEGG database shows that it involves oxidative phosphorylation,myocardial contraction,PPAR signaling pathway,peroxisomes,and RIG-I-like receptor signaling pathway.Conclusion:The detection results of the chip revealed the unique lncRNA expression profile of the atherosclerosis model.The differential lncRNAs and mRNAs were mostly up-regulated.Among them,the super-conservative sequence uc.84 was highly expressed for the next study;GO analysis showed that the differentially expressed lncRNA and metabolism Related to inflammation,Pathway analysis is related to inflammatory signaling pathways such as PPAR signaling pathway,TNF signaling pathway,CAM and NOD-like receptor signaling pathway.These inflammatory pathways are now widely studied and proven to be associated with atherosclerosis.Part ? Effect of long non-coding RNA uc.84 on vascular smooth muscle cellsObjective:In order to evaluate the role of uc.84 in the proliferation and migration of VSMC,we conducted siRNA-mediated knockdown experiments.Methods:The expression specificity of uc.84 cells was identified by qRT-PCR,the expression of uc.84 was evaluated after PDGF-BB stimulation,VSMCs were cultured in vitro and divided into 4 groups,blank control group;PDGF-BB group(concentration 20ng/mL);siRNA intervention group;siRNA+PDGF-BB group(concentration:20ng/mL);Western Blot detection and scratch test were performed respectivelyResults:1.Uc.84 is enriched in vascular smooth muscle cells,and after PDGF-BB stimulation,it is down-regulated in a concentration and time-dependent manner within a certain range2.Compared with the control group,the expression of PCNA was up-regulated after siRNA transfection,and the expression of contractile phenotype protein ?-SMA was down-regulated(P<0.05),indicating that siRNA uc.84 transfection of VSMC can mimic the effect of PDGF-BB,The phenotype of smooth muscle cells from normal contraction state to proliferation state,and uc.84 silencing does not reduce the effect of PDGF-BB.Conclusion:lncRNA uc.84 plays an important role in maintaining the normal contraction phenotype of blood vessels,can inhibit the proliferation and migration of vascular smooth muscle cells,and plays a protective role in the formation of atherosclerosis.Part ? Circulating blood uc.84 early diagnosis of acute myocardial infarctionObjective:Explore whether uc.84 can be used as an independent diagnostic factor for acute myocardial infarction and the accuracy of early diagnosis of acute myocardial infarction.Methods:The difference in expression of uc.84 was detected in whole blood specimens from 42 patients with acute myocardial infarction and 42 non-acute myocardial infarction controls,and ROC curve was used to judge the diagnostic reliability of uc.84 as a biomarker for acute myocardial infarction.Results:1.Comparing the baseline clinical characteristics of subjects,the cardiac biomarkers cTnT and CK-MB were significantly different between AMI and non-AMI patients(P<0.001),and the difference between white blood cell counts was statistically significant;2.The room temperature standing time has almost no effect on the plasma level of uc.84,and there is no obvious correlation between uc.84 and white blood cell count.3.Compared with non-AMI patients,the expression of uc.84 in whole blood of AMI patients was significantly increased.The area under the ROC curve(AUC)of UC.84 diagnosis of AMI was 0.8588(95%confidence interval),when its relative expression level At the optimal cutoff point,the specificity was 73.81%and the sensitivity was 83.33%.Conclusion:uc.84 has a significant difference in expression between patients with acute myocardial infarction and non-acute myocardial infarction,and is used to diagnose AMI with high specificity and sensitivity,and can provide a reference for AMI diagnosis. |
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