Study On The Mechanism Of MiRNA-223 Involved In Platelet Derived Microbubble In Mice Atherosclerosis

BackgroundMicrobubble mainly exists in platelets,microRNAs contains protein and other bioactive substances involved in signal transduction between cells.A large number of studies show that in atherosclerosis and its complications in the patient's blood,content of microbubbles significantly increased.MicroRNAs is widely in the study of gene expression regulation factor,the structure is about the 22 ~ 25 nucleotide single stranded RNA molecules.In recent years,the important role of miRNAs in cardiovascular system has been elucidated,has made great achievements.Our previous study found that platelet is activated in the future,The content of the secretion of microbubbles of miRNAs in change,and can be absorbed in endothelial cells,thus affecting the proliferation,migration,apoptosis and secretion.In the process,change the content of miRNA-223 was most significant.With the progress of the study,and role of miRNA-223 in the process of atherosclerosis more attention mechanism more and more clear,there may be a new biological targets for diagnosis and treatment of atherosclerosis.Objective1 C57BL/6 pure ApoE gene knockout mice as the basic research object,the establishment of appropriate animal model and grouping,in order to explore the PMP miRNA-223 provides powerful and reliable basis of the animal experiment in the repair mechanisms of atherosclerotic endothelial injury.2 The serum lipid levels and the miRNA-223 content in the platelet derived microbubbles were compared between the two groups.3 To analyze the degree of atherosclerosis in each group of mice,and to explore the relationship between the degree of atherosclerosis and the miRNA-223 content in platelet derived microbubbles.Methods1 The apolipoprotein E gene about 6 week old C57BL/6 inbred background knockout male mice(ApoE-/-)and 12 week old C57BL/6 6 male mice as the research object,divided into three groups under the same feeding condition,the high-fat diet ApoE-/-mice group,intravenous injection miRNA-223 antagomir inhibits the transcriptional expressionof miRNA-223(AG group);a high-fat diet,were injected into the tail vein of ApoE-/-mice with the same dose of normal saline group(AE group);give full nutrient material in SPF The mice were injected with the same dose of normal saline by tail vein(control group).2 Raised up to 12 weeks after the three groups of mice were weighed,sacrificed,blood centrifugation supernatant,using automatic biochemical analyzer to detect the serum lipid levels in mice;extraction of mouse blood platelets microbubbles,determination of PMP miRNA-223 real-time PCR.3 Of mice aorta atherosclerosis were gross observation,HE staining under optical microscope and pathological observation of medical digital image analysis system to produce the thickness of plaque area,were quantitatively analyzed.Results1 Since the purchase of the mice fed to disposal before death,18 mice survived,no signs of life,death,disease,anorexia and other adverse conditions.On three groups of mice were weighed,group AG(27.21 + 2.05)g,AE group(27.72 + 2.59)g control.Group(23.51 + 1.55)g.weighing results showed that the body weight of AG group and AE group were no statistically significant difference(P>0.05),AG group and control group,AE group and control group,there were significant differences in body weight(P<0.05,P<,AG two,AE 0.05)The body weight of the mice was higher than that of the control group.2 Through gradient centrifugation,ultracentrifugation,reverse transcription and quantitative PCR method to detect the relative level of comparison,three groups of miRNA-223 mice in PMP showed that the miRNA-223 content of AG group in PMP mice than in the other two groups were significantly decreased,while the AE group was higher than that in control group.Converted into multiple display: AG group and control group was(0.14 + 0.06vs1.02 + 0.09,P<0.05;AG)and group AE(0.14 + 0.06vs2.71 + 0.4 4,P<0.05),AE group and control group(2.71 + 0.44vs1.02 + 0.09,P<0.05).The detection results of blood lipid level of mice showed that mice in three groups TG,TC,LDL-C,HDL-C levels were divided into AG group(0.60 + 0.08mmol/L,11.03 + 1.29 + 0.86mmol/L,5.06 mmol/L,0.75 0.99 + mmol/L),group AE(0.61 + 0.10 mmol/L 11.07 + 1.53Mmol/L,5.13 + 1.13 mmol/L 0.70 + 0.99 mmol/L),control group(0.34 + 0.70 + 0.54mmol/L,3.18 mmol/L,1.59 mmol/L + 0.36,2.28 + 0.37 mmol/L)of.AG group and AEgroup were TG,TC,LDL-C and HDL-C levels were not significantly different(P>0.05),compared with the control group AG group and AE group of TG,TC and LDL-C levels were significantly higher and statistically significant The difference(P<0.05,P<0.05),HDL-C level was significantly lower than the control group and the difference was statistically significant(P<0.05,P<0.05).3 The mouse aortic atherosclerotic lesions and observed under optical microscope to observe the pathological changes,AG group and AE group were almost all different degrees of atherosclerosis,while the control group were almost without atherosclerotic change.Analysis system of AG group use of medical digital images,atherosclerotic plaque group mice AE thickness and plaque area were quantitatively analyzed show that the small group of two In the plaque thickness and size were: AG group:(0.095 + 0.013)mm,(0.075 + 0.015)mm2;AE group:(0.104 + 0.015)mm,(0.080 + 0.014)thickness and plaque area between mm2.AG group and AE group of atherosclerotic plaque were not statistically significant(P>0.05,P> 0.05)the AG group and AE group.All mice(n=12)the relative content of miRNA-223 and plaque area in the corresponding PMP in the correlation between the results.Display: AG group and AE group compared the correlation between them(r=0.22,P>0.05),AG group(r=0.36,P>0.05),AE group(r=0.06,P>0.05)there was no significant correlation between.PMP miRNA-223 AS and the relative level of plaque area.Conclusion1 The mice in the three groups were able to survive,and the AG group and the AE group had different degrees of atherosclerosis through high fat diet.2 The content of miRNA-223 in group AE atherosclerosis PMP mice than in normal control mice increased;miRNA-223 transcription and synthesis can be inhibited by miRNA-223 antagomir,the content of miRNA-223 AG mice in PMP group was significantly lower than that of AE group and control group of mice.3 AG group and AE group of mice in weight,TC,TG,LDL-C,the degree of atherosclerotic lesions were higher than the control group,HDL-C was lower than the control group;the weight between AG group and AE group of mice,no link to start targeting regulatory miRNA-223.PMP significant difference on serum lipid level in the ApoE may.PMP miRNA223 may be the main involved in atherosclerosis lipid levels and atherosclerosis,in has occurred AG group and AE group mice atherosclerosis,the severityof atherosclerosis in MI and PMP There was no significant correlation between the content of RNA-223.