Chinese Journal Of Cell And Stem Cell >Archive >2018 >Volume 8 Number 6 Dec 2018 >Original Researches >contents

Journal content

Mechanism of circulating endothelial microparticles related microRNA in the atherosclerosis induced by inflammatory response of macrophage

Chen Hongying, Zhang Yuhang, Gao Guangmin, et al


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【Abstract】 Objective To study the circulating endothelial microparticles related microRNA in the atherosclerosis induced by inflammatory response of macrophage. Methods Five-week-old mice of SPF-grade were randomized to an experimental group and a control group (n = 50 for each group). The experimental mice were divided into four groups: a control group, an atherosclerosis (AS) model group, an NC-microRNA group and a microRNA-19 b inhibitor group. The histology of atherosclerotic lesions was observed with oil red staining, and the plaque area ratio was calculated. Serum cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C), TNF-α,IL-1, IL-6 and IL-10 were measured. The expressions of apoptosis related proteins Bcl-2,cleaved-caspase-3 and Bax were detected with Western Blot,the mir-19b expression was detected with real-time quantitative PCR,and the apoptosis rate of macrophages was detected with flow cytometry. Then the comparison among groups were analyzed with variance by single-factor or repeated measures, the comparison between two groups were analyzed with LSD-t test. Results The percentage of plaque area of thoracic artery in each group: (0.00±0.00)﹪ in the control group, (9.59±6.53)﹪ in the AS model group, (8.96±3.47)﹪in the NC-microRNA group and (3.21±2.03)﹪ in the microRNA19b inhibitor group,and the differences of the three group were statistically significant (F = 20.572, P = 0.002). Compared with the control group, TC (TC) in the AS model group and the NC-microRNA group were as follows: TC[AS model group (3.26±0.21) mmol/L; NC-microRNA group (3.13±0.14) mmol/ L; F = 13.994, P = 13.994, P = 0.002], TG [AS model group 0.25 (0.25±0.06) mmol/ L; AS model group (1.65±0.11) mmol/L; NC-miRNA (1.59±0.27) mmol/L; F = 10.069, P = 0.006) increased, and HDL-C (AS) increased. In the model group, the levels of NC-microRNA (0.08±0.09) mmol/ L, NC-microRNA (0.08±0.05) mmol/L and F = 12.450, P = 0.004 decreased. Compared with the model group, the levels of TC (1.85±0.06) mmol/L, TG (0.15±0.03) mmol/ L and LDL-C (1.21±0.10) mmol/L decreased, while HDL-C (0.11±0.05) mmol/L increased in the miRNA- 19b inhibitor group (P < 0.05). Compared with the control group, the levels of IL-1 (34.06±3.58) g/ L, IL-6 (92.57±31.97) g/Land TNF-alpha (63.01±15.65)g/L increased, while the levels of IL-10 (16.86±1.29) g/L decreased, and the indexes of NC-microNA group also showed a same change trend; the levels of IL-1(24.85±6.21) g/L, IL-6 (53.29±17.15) g/L and TNF-alpha (34.51±6.47) g/ L in the inhibitor group of microRNA-19b were lower than those in the control group. The levels of IL- 10 (24.94±4.72 )g/L in Group A and NC-microRNA (F = 13.671, P = 0.002) were higher than those in Group B (F = 13.675, P = 0.008). Compared with the control group, Bax (AS model group 3.39±0.01; AS model group 3.39±0.01; NC- microRNA group 0.64 ±0.02; F = 26.910, P = 0.000) and cleaved-PARP (AS model group 2.47±0.05; NANC-microRNA group 3.27±0.01; F = 13.226, P = 0.000) expression increased. However, Bcl-2 (model group 0.67±0.02; NC-microRNA group 0.64±0.02; F = 12.585, P = 0.000) expression level was down-regulated. Compared with the NC-microRNA group, the expression levels of Bax (2.16±0.02) and cleaved-PARP (1.91±0.04) were down-regulated, while the expression levels of Bcl-2 (1.05±0.01) were up-regulated. Compared with the control group, the transcription level of microRNA-19b in the AS model group (2.71±0.02) and NC-microRNA group (2.43±0.02) D increased significantly (P < 0.05). The transcription level of the microRNA-19b inhibitor group (1.52±0.01) increased, but the increase was not significant (F = 15.353, P = 0.002) compared with the AS model group and NC-microRNA group. The transcription level of microRNA-19b gene was 1.02±0.03 in the control group, 2.71±0.02 in the AS model group, 2.43±0.02 in the NC-microRNA group and 1.52±0.01 in the microRNA-19b inhibitor group (P < 0.05). Apoptotic rate: (4.41±0.18)﹪ in thecontrol group, (7.16±0.73)﹪ in the AS model group, (6.29±0.24)﹪ in the NC-microRNA group and (5.01±0.11)﹪ in the microRNA19b inhibitor group, (F = 12.889, P = 0.008). Conclusion The inhibitor microRNA-19b can reduce macrophage apoptosis, decrease the pro-inflammatory factors,and reduce the development of blood vessel thickness caused by atherosclerosisi. It also can decrease the lipid TC, TG  and LDL-C and increase the HDL-C,so that it may have active guiding significance for the clinical diagnosis and treatment of coronary heart disease.
【Key words】 Endothelial particles; Micrornas-19b; Macrophages; Inflammat-ory factors; Atherosclerosisi