miR-423、miR-18b、miR-129在老龄小鼠心衰过程中的表达
发布时间：2018-08-23 18:55
【摘要】：背景及目的随着人口老龄化,心力衰竭(简称心衰)基础疾病(冠心病、高血压病、心肌梗死等)的发病率逐年升高,心衰的发病率及死亡率也随之不断提高。寻找一种既能诊断心衰又能较好反映其严重程度的检测指标有着重要的临床意义。心衰发展过程中伴随着多种结构的改变,如心肌细胞肥大、凋亡、间质纤维化、毛细血管数量减少及免疫系统的激活等,结构的改变引起心脏功能的变化(如射血量降低、充盈压增高等),但是潜在的分子机制尚未阐明。微小RNA(mi RNAs)是具有21-25个核苷酸的内源性小分子RNA。近年来,大量研究表明血清中的mi RNAs已作为多种组织损伤及病理过程的敏感和特异的生物标志物。mi RNAs通过多种心脏病理改变引起心肌重构、心功能降低,最终导致心力衰竭。因此,我们可以通过一些特定的mi RNAs水平来诊断心力衰竭。本研究包括:1)建立老龄小鼠心衰模型,观察老龄小鼠心衰过程中mi R-423、mi R-18b、mi R-129的表达差异。2)研究老龄小鼠在心衰发生发展过程中,mi R-423的表达趋势,探讨其与心衰的关系,为心衰的诊断或预后提供价值。方法1.18月龄雄性健康小鼠共54只,按照投硬币方法随机分成2组。心衰组(n=38):常规饲养,于腹部皮下注射异丙肾上腺素(ISO)持续14天,构造老龄小鼠心力衰竭模型;对照组(n=16):常规饲养,相同方法注射等量生理盐水14天。注射后继续常规饲养4周。2.4周之后行超声心动图和心电图,比较两组心脏结构、功能和心率。然后处死小鼠10只(心衰4w组),取心脏标本,分别给予全心及左室称重,计算出左室质量指数(LVMI)。心肌标本经固定、脱水、包埋后,切得组织切片,分别给予常规HE染色。3.继续饲养2周后处死小鼠10只(心衰6w组),剩余小鼠继续饲养2周后处死(心衰8w组)。处死后按压小鼠的眼球取血4ml,离心后收集上清液。4.从血浆中提取总RNA并进行逆转录反应,获得相应mi RNA的c DNA,以β-actin为内参进行标定,采用实时荧光定量PCR(RT-PCR)技术检测mi R-423、mi R-18b、mi R-129在心衰小鼠及正常小鼠血清中的表达。5.用PCR检测mi R-423在各组血清中表达量,并进行对比。6.应用SPSS 16.0软件包进行数据处理,计数资料用均数±标准差(sx±)表示,计量资料若满足正态性,采用t检验,若不满足正态性,采用秩和检验;多组比较用Kruskal-Wallis H检验,P0.05为差异有统计学意义。结果1.左室质量指数(LVMI,mg/g)比较:对照组为2.68±0.32,心衰组为5.70±0.54。与对照组比较,心衰组左室质量指数明显升高,差异有统计学意义(P0.01)。2.HE染色:正常对照组小鼠的心肌细胞轮廓完整,肌纤维布列一致;心衰组小鼠心肌纤维部分中断,心肌细胞呈现不同程度的水肿、心肌细胞肥大,有局灶性或者片状坏死灶。3.心脏彩超:心衰组心室腔明显增大,心率较快,左室短轴收缩功能障碍更明显(P0.05)。4.心电图:与对照组相比,心衰组心率较快,差异有统计学意义(P0.05)。5.mi R-423、mi R-18b、mi R-129表达量比较:心衰小鼠血清中mi R-423表达显著高于对照组(P0.001),mi R-18b、mi R-129的表达与对照组相比,差异无统计学意义(P0.05)。6.心衰后不同时间,PCR检测发现mi R-423的表达量呈上升趋势,差异有统计学意义(P0.01)。结论1.与对照组相比,心衰组中mi R-423表达上调,而mi R-18b、mi R-129的表达差异无统计学意义。其可能作为一种血清中潜在的新型生物标志物,对将来心衰的诊断起重要作用。2.在心衰发生发展过程中,mi R-423的表达量逐渐升高。其对将来心衰严重程度的评估起重要作用。
[Abstract]:BACKGROUND & OBJECTIVE With the aging of the population, the incidence of basic diseases of heart failure (coronary heart disease, hypertension, myocardial infarction, etc.) is increasing year by year, and the incidence and mortality of heart failure are also increasing. The development of heart failure is accompanied by many structural changes, such as cardiomyocyte hypertrophy, apoptosis, interstitial fibrosis, reduction of capillary number and activation of the immune system. The structural changes lead to changes in cardiac function (such as decreased ejection and increased filling pressure), but the underlying molecular mechanisms are not yet understood. MicroRNAs (mi RNAs) are 2. Endogenous small RNA of 1-25 nucleotides. In recent years, a large number of studies have shown that serum mi RNAs have been used as sensitive and specific biomarkers for various tissue damage and pathological processes. This study included: 1) Establish the heart failure model of aged mice, observe the expression of MIR-423, MIR-18b, MIR-129 in the heart failure process of aged mice. 2) Study the expression trend of MIR-423 in the heart failure process of aged mice, and explore the relationship between MIR-423 and heart failure. Methods Fifty-four male healthy mice aged 1.18 months were randomly divided into two groups according to the coin-dropping method. Heart failure group (n=38): routine feeding, subcutaneous injection of isoproterenol (ISO) for 14 days to construct the heart failure model of aged mice; control group (n=16): routine feeding, injecting the same amount of saline for 14 days. After 4 weeks of routine feeding, echocardiography and electrocardiogram were performed to compare the cardiac structure, function and heart rate of the two groups. Then 10 mice were sacrificed and weighed with whole heart and left ventricle respectively. LVMI was calculated. E staining. 3. After 2 weeks of continuous feeding, 10 mice were killed (heart failure group 6 w), and the remaining mice were killed after 2 weeks of continuous feeding (heart failure group 8 w). After death, the eyeballs of mice were pressed for 4 ml, and the supernatant was collected after centrifugation. 4. Total RNA was extracted from plasma and reverse transcription reaction was performed to obtain the corresponding C DNA of MI RNA, which was calibrated with beta-actin as internal reference, and real-time was used. The expression of MIR-423, MIR-18b and MIR-129 in serum of HF mice and normal mice was detected by fluorescence quantitative PCR (RT-PCR). 5. The expression of MIR-423 in serum of HF mice and normal mice was detected by PCR and compared. 6. The data were processed by SPSS 16.0 software package. The counting data were expressed by mean (+) standard deviation (sx (+). Results 1. Left ventricular mass index (LVMI, mg/g) was significantly higher in the heart failure group than in the control group (2.68 [0.32], and 5.70 [0.54] in the heart failure group. (P 0.01). 2. HE staining: the normal control group of Mice Myocardial Cells contour is intact, the arrangement of myocardial fibers is consistent; heart failure group of mice myocardial fibers partially interrupted, myocardial cells showed varying degrees of edema, hypertrophy of myocardial cells, focal or patchy necrosis foci. ECG: Compared with the control group, the heart rate of HF group was faster, the difference was statistically significant (P 0.05). 5. The expression of MIR-423, MIR-18b, MIR-129 in the serum of HF mice was significantly higher than that of the control group (P 0.001), and the expression of MIR-18b, MIR-129 in the serum of HF mice was not statistically significant (P 0.05). Conclusion 1. Compared with the control group, the expression of MIR-423 was up-regulated in HF group, but the expression of MIR-18b and MIR-129 was not significantly different. It may be a potential new biomarker in serum, and it may be a potential biomarker for future heart disease. 2. The expression of MIR-423 increased gradually during the development of heart failure. It plays an important role in the evaluation of the severity of heart failure in the future.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R541.6
本文编号：2199607
[Abstract]:BACKGROUND & OBJECTIVE With the aging of the population, the incidence of basic diseases of heart failure (coronary heart disease, hypertension, myocardial infarction, etc.) is increasing year by year, and the incidence and mortality of heart failure are also increasing. The development of heart failure is accompanied by many structural changes, such as cardiomyocyte hypertrophy, apoptosis, interstitial fibrosis, reduction of capillary number and activation of the immune system. The structural changes lead to changes in cardiac function (such as decreased ejection and increased filling pressure), but the underlying molecular mechanisms are not yet understood. MicroRNAs (mi RNAs) are 2. Endogenous small RNA of 1-25 nucleotides. In recent years, a large number of studies have shown that serum mi RNAs have been used as sensitive and specific biomarkers for various tissue damage and pathological processes. This study included: 1) Establish the heart failure model of aged mice, observe the expression of MIR-423, MIR-18b, MIR-129 in the heart failure process of aged mice. 2) Study the expression trend of MIR-423 in the heart failure process of aged mice, and explore the relationship between MIR-423 and heart failure. Methods Fifty-four male healthy mice aged 1.18 months were randomly divided into two groups according to the coin-dropping method. Heart failure group (n=38): routine feeding, subcutaneous injection of isoproterenol (ISO) for 14 days to construct the heart failure model of aged mice; control group (n=16): routine feeding, injecting the same amount of saline for 14 days. After 4 weeks of routine feeding, echocardiography and electrocardiogram were performed to compare the cardiac structure, function and heart rate of the two groups. Then 10 mice were sacrificed and weighed with whole heart and left ventricle respectively. LVMI was calculated. E staining. 3. After 2 weeks of continuous feeding, 10 mice were killed (heart failure group 6 w), and the remaining mice were killed after 2 weeks of continuous feeding (heart failure group 8 w). After death, the eyeballs of mice were pressed for 4 ml, and the supernatant was collected after centrifugation. 4. Total RNA was extracted from plasma and reverse transcription reaction was performed to obtain the corresponding C DNA of MI RNA, which was calibrated with beta-actin as internal reference, and real-time was used. The expression of MIR-423, MIR-18b and MIR-129 in serum of HF mice and normal mice was detected by fluorescence quantitative PCR (RT-PCR). 5. The expression of MIR-423 in serum of HF mice and normal mice was detected by PCR and compared. 6. The data were processed by SPSS 16.0 software package. The counting data were expressed by mean (+) standard deviation (sx (+). Results 1. Left ventricular mass index (LVMI, mg/g) was significantly higher in the heart failure group than in the control group (2.68 [0.32], and 5.70 [0.54] in the heart failure group. (P 0.01). 2. HE staining: the normal control group of Mice Myocardial Cells contour is intact, the arrangement of myocardial fibers is consistent; heart failure group of mice myocardial fibers partially interrupted, myocardial cells showed varying degrees of edema, hypertrophy of myocardial cells, focal or patchy necrosis foci. ECG: Compared with the control group, the heart rate of HF group was faster, the difference was statistically significant (P 0.05). 5. The expression of MIR-423, MIR-18b, MIR-129 in the serum of HF mice was significantly higher than that of the control group (P 0.001), and the expression of MIR-18b, MIR-129 in the serum of HF mice was not statistically significant (P 0.05). Conclusion 1. Compared with the control group, the expression of MIR-423 was up-regulated in HF group, but the expression of MIR-18b and MIR-129 was not significantly different. It may be a potential new biomarker in serum, and it may be a potential biomarker for future heart disease. 2. The expression of MIR-423 increased gradually during the development of heart failure. It plays an important role in the evaluation of the severity of heart failure in the future.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R541.6
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1 何芳;miR-423、miR-18b、miR-129在老龄小鼠心衰过程中的表达[D];郑州大学;2015年
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