本文選題：低氧性肺動脈高壓 + 循環(huán)miRNAs��； 參考：《石河子大學(xué)》2015年碩士論文

【摘要】：研究目的的::模擬6,000米海拔低壓低氧條件,建立低壓低氧誘發(fā)肺動脈高壓(HPH)的大鼠模型,研究低壓低氧暴露條件下HPH模型大鼠循環(huán)mi RNA表達(dá)譜特征及其與HPH病理變化的相關(guān)性,探索循環(huán)miRNA作為HPH診斷生物標(biāo)志物的可行性。研究方法:1.隨機(jī)將120只雄性6周齡SD大鼠分為低氧4d、7d、10d、14d和21d暴露組及各應(yīng)時點(diǎn)對照組,置人工氣候艙常規(guī)飼養(yǎng),繼以模擬6,000m海拔進(jìn)行低壓低氧暴露,按分組時點(diǎn)定時采用直接測壓法檢測平均肺動脈壓(m PAP)并采集血液及組織標(biāo)本,心肺組織標(biāo)本經(jīng)固定、包埋后進(jìn)行HE染色及病理分析。2.依據(jù)低壓低氧后m PAP的變化,取低氧14d暴露組及對照組大鼠血清標(biāo)本,利用基因芯片技術(shù)進(jìn)行循環(huán)mi RNA表達(dá)譜檢測,不同組別間循環(huán)mi RNA表達(dá)譜差異分析采用聚類分析。3.利用熒光定量PCR技術(shù)對候選循環(huán)mi RNA進(jìn)行驗(yàn)證,采用相對定量法進(jìn)行數(shù)據(jù)分析。研究結(jié)果:1.低壓低氧條件暴露10d、14d、21d的SD大鼠m PAP、右心室肥厚指數(shù)(RVHI)及肺小動脈血管壁厚占外徑比值(MT%)較同時點(diǎn)對照組顯著升高(P0.01)。2.基因芯片結(jié)果顯示,累計從大鼠血清中檢測到723種循環(huán)mi RNAs,按照FC2、Flag/Call值3及P0.05的標(biāo)準(zhǔn),從低氧14d組及對照組大鼠間篩選出6種差異化表達(dá)的mi RNAs,其中低氧組中表達(dá)上調(diào)的mi RNA有3種(let-7d、mi R-451、mi R-505),表達(dá)下調(diào)的mi RNA有3種(mi R-455-3p、mi R-455-5P、mi R-214)。3.熒光定量PCR定量驗(yàn)證結(jié)果顯示,與對照組相比,低氧14d組大鼠血清中mi R-451、mi R-505、let-7d表達(dá)上調(diào)(P0.01),mi R-214表達(dá)下調(diào)(P0.05),mi R-455和mi R-455-5p表達(dá)與對照組相比無顯著差異;ROC曲線表明,mi R-451、mi R-505及l(fā)et-7d的用于鑒別低氧性HPH大鼠和對照組大鼠的曲線下面積(AUC)均大于0.8,且與機(jī)體脂質(zhì)過氧化指標(biāo)丙二醛(MDA)含量均正相關(guān)。結(jié)論:模擬海拔6000m高原環(huán)境低壓低氧14d即可成功建立低壓低氧誘發(fā)肺動脈高壓的大鼠模型;基于大鼠模型,獲得了低壓低氧暴露條件下HPH模型大鼠循環(huán)mi RNA表達(dá)譜,初步發(fā)現(xiàn)4種循環(huán)mi RNA在HPH大鼠和對照組大鼠間存在顯著性差異化表達(dá),可能與HPH病理變化相關(guān)。
[Abstract]:The purpose of this study is to simulate the hypobaric hypoxia conditions of 6000 m, establish a rat model of pulmonary hypertension (HPH) induced by low pressure hypoxia, and study the characteristics of MI RNA expression in HPH model rats under hypobaric hypoxia exposure and the correlation with the pathological changes of HPH, and explore the feasibility of circulating miRNA as a biomarker for HPH diagnosis. Method: 1. the 120 male 6 week old SD rats were randomly divided into hypoxia 4D, 7d, 10d, 14d and 21d exposure group and each time point control group, and the artificial climate cabin was routinely raised. The average pulmonary arterial pressure (m PAP) was detected by direct pressure measurement at the time point of the group at the time point of the group, and the blood and tissue specimens were collected, and the blood and tissue specimens were collected and the heart and lung were collected. The tissue specimens were fixed, embedded after HE staining and pathological analysis,.2. based on the changes of M PAP after hypobaric hypoxia, the serum samples of hypoxia 14d exposure group and control group were taken, the expression of circulating mi RNA expression was detected by gene chip technology. The difference analysis of RNA expression spectrum of circulating mi between different groups was analyzed by cluster analysis.3. using fluorescence quantitative PCR. A relative quantitative method was used to verify the candidate cycle mi RNA. The results were as follows: 1. the low oxygen and hypoxia conditions exposed 10d, 14d, 21d SD rats, m PAP, the right ventricular hypertrophy index (RVHI) and the ratio of the diameter of the pulmonary artery wall thickness (MT%) significantly higher than those in the same group (P0.01).2. gene chip results showed the cumulative from In the rat serum, 723 kinds of circulating mi RNAs were detected. According to the standard of FC2, Flag/Call value 3 and P0.05, 6 kinds of differential expression mi RNAs were screened from the hypoxia 14d group and the control group, of which there were 3 kinds of MI RNA in the hypoxia group. There were 3 kinds of downregulation. Compared with the control group, the serum mi R-451, MI R-505, let-7d expression up regulated (P0.01) and MI R-214 expression decreased (P0.05) compared with the control group, and there was no significant difference in the expression of MI R-214 expression between the 14d and the control group. The area under the curve (AUC) of the control group was more than 0.8, and it was positively correlated with the content of malondialdehyde (MDA) in the lipid peroxidation index of the body. Conclusion: the rat model of pulmonary hypertension induced by hypobaric hypoxia at the altitude of 6000m plateau can be successfully established. Based on the rat model, the HPH model under the condition of low pressure and hypoxia exposure is obtained. The expression of MI RNA in rats circulated, and it was found that there was a significant difference in the expression of 4 kinds of circulating mi RNA between the HPH rats and the control rats, which may be related to the pathological changes of HPH.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R544.1;R440

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本文編號：1955880