miR-423、miR-18b、miR-129在老齡小鼠心衰過程中的表達
發(fā)布時間：2018-08-23 18:55
【摘要】：背景及目的隨著人口老齡化,心力衰竭(簡稱心衰)基礎(chǔ)疾病(冠心病、高血壓病、心肌梗死等)的發(fā)病率逐年升高,心衰的發(fā)病率及死亡率也隨之不斷提高。尋找一種既能診斷心衰又能較好反映其嚴(yán)重程度的檢測指標(biāo)有著重要的臨床意義。心衰發(fā)展過程中伴隨著多種結(jié)構(gòu)的改變,如心肌細(xì)胞肥大、凋亡、間質(zhì)纖維化、毛細(xì)血管數(shù)量減少及免疫系統(tǒng)的激活等,結(jié)構(gòu)的改變引起心臟功能的變化(如射血量降低、充盈壓增高等),但是潛在的分子機制尚未闡明。微小RNA(mi RNAs)是具有21-25個核苷酸的內(nèi)源性小分子RNA。近年來,大量研究表明血清中的mi RNAs已作為多種組織損傷及病理過程的敏感和特異的生物標(biāo)志物。mi RNAs通過多種心臟病理改變引起心肌重構(gòu)、心功能降低,最終導(dǎo)致心力衰竭。因此,我們可以通過一些特定的mi RNAs水平來診斷心力衰竭。本研究包括:1)建立老齡小鼠心衰模型,觀察老齡小鼠心衰過程中mi R-423、mi R-18b、mi R-129的表達差異。2)研究老齡小鼠在心衰發(fā)生發(fā)展過程中,mi R-423的表達趨勢,探討其與心衰的關(guān)系,為心衰的診斷或預(yù)后提供價值。方法1.18月齡雄性健康小鼠共54只,按照投硬幣方法隨機分成2組。心衰組(n=38):常規(guī)飼養(yǎng),于腹部皮下注射異丙腎上腺素(ISO)持續(xù)14天,構(gòu)造老齡小鼠心力衰竭模型;對照組(n=16):常規(guī)飼養(yǎng),相同方法注射等量生理鹽水14天。注射后繼續(xù)常規(guī)飼養(yǎng)4周。2.4周之后行超聲心動圖和心電圖,比較兩組心臟結(jié)構(gòu)、功能和心率。然后處死小鼠10只(心衰4w組),取心臟標(biāo)本,分別給予全心及左室稱重,計算出左室質(zhì)量指數(shù)(LVMI)。心肌標(biāo)本經(jīng)固定、脫水、包埋后,切得組織切片,分別給予常規(guī)HE染色。3.繼續(xù)飼養(yǎng)2周后處死小鼠10只(心衰6w組),剩余小鼠繼續(xù)飼養(yǎng)2周后處死(心衰8w組)。處死后按壓小鼠的眼球取血4ml,離心后收集上清液。4.從血漿中提取總RNA并進行逆轉(zhuǎn)錄反應(yīng),獲得相應(yīng)mi RNA的c DNA,以β-actin為內(nèi)參進行標(biāo)定,采用實時熒光定量PCR(RT-PCR)技術(shù)檢測mi R-423、mi R-18b、mi R-129在心衰小鼠及正常小鼠血清中的表達。5.用PCR檢測mi R-423在各組血清中表達量,并進行對比。6.應(yīng)用SPSS 16.0軟件包進行數(shù)據(jù)處理,計數(shù)資料用均數(shù)±標(biāo)準(zhǔn)差(sx±)表示,計量資料若滿足正態(tài)性,采用t檢驗,若不滿足正態(tài)性,采用秩和檢驗;多組比較用Kruskal-Wallis H檢驗,P0.05為差異有統(tǒng)計學(xué)意義。結(jié)果1.左室質(zhì)量指數(shù)(LVMI,mg/g)比較:對照組為2.68±0.32,心衰組為5.70±0.54。與對照組比較,心衰組左室質(zhì)量指數(shù)明顯升高,差異有統(tǒng)計學(xué)意義(P0.01)。2.HE染色:正常對照組小鼠的心肌細(xì)胞輪廓完整,肌纖維布列一致;心衰組小鼠心肌纖維部分中斷,心肌細(xì)胞呈現(xiàn)不同程度的水腫、心肌細(xì)胞肥大,有局灶性或者片狀壞死灶。3.心臟彩超:心衰組心室腔明顯增大,心率較快,左室短軸收縮功能障礙更明顯(P0.05)。4.心電圖:與對照組相比,心衰組心率較快,差異有統(tǒng)計學(xué)意義(P0.05)。5.mi R-423、mi R-18b、mi R-129表達量比較:心衰小鼠血清中mi R-423表達顯著高于對照組(P0.001),mi R-18b、mi R-129的表達與對照組相比,差異無統(tǒng)計學(xué)意義(P0.05)。6.心衰后不同時間,PCR檢測發(fā)現(xiàn)mi R-423的表達量呈上升趨勢,差異有統(tǒng)計學(xué)意義(P0.01)。結(jié)論1.與對照組相比,心衰組中mi R-423表達上調(diào),而mi R-18b、mi R-129的表達差異無統(tǒng)計學(xué)意義。其可能作為一種血清中潛在的新型生物標(biāo)志物,對將來心衰的診斷起重要作用。2.在心衰發(fā)生發(fā)展過程中,mi R-423的表達量逐漸升高。其對將來心衰嚴(yán)重程度的評估起重要作用。
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R541.6
本文編號：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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R541.6
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1 何芳;miR-423、miR-18b、miR-129在老齡小鼠心衰過程中的表達[D];鄭州大學(xué);2015年
，本文編號：2199607
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