發(fā)布時(shí)間：2018-01-22 05:24

  本文關(guān)鍵詞： 肺動(dòng)脈高壓 TGF-β1 miRNA-19a 小鼠肺動(dòng)脈高壓模型　出處：《西南醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:本研究通過(guò)小鼠動(dòng)力性肺動(dòng)脈高壓模型探索肺組織TGF-β1及相關(guān)mi RNAs的表達(dá)與肺血管重構(gòu)及肺動(dòng)脈高壓形成之間的關(guān)系,進(jìn)而了解TGF-β1及相關(guān)mi RNAs的表達(dá)是否與肺動(dòng)脈高壓的進(jìn)展快慢有相關(guān)性,從而提供早期診斷CHD-PAH(先天性心臟病相關(guān)肺動(dòng)脈高壓)的臨床指標(biāo)。方法:建立肺動(dòng)脈高壓小鼠模型,以模型小鼠肺組織為研究對(duì)象,進(jìn)一步驗(yàn)證血流動(dòng)力學(xué)的改變對(duì)肺組織TGF-β1及相關(guān)miRNAs表達(dá)的影響。方法如下:將20只雄性C57小鼠隨機(jī)分成2組:A1,A2,其中A1為實(shí)驗(yàn)組,A2為假手術(shù)組。兩組均采用戊巴比妥鈉以40mg/kg腹腔麻醉。實(shí)驗(yàn)組左側(cè)第3肋間開(kāi)胸,切除左肺,假手術(shù)組左側(cè)第3肋間開(kāi)胸,不切除左肺。手術(shù)8周后處死,實(shí)驗(yàn)組、假手術(shù)組分別測(cè)右心室收縮壓(RVSP)及稱量右心室(RV)和左心室(LV)+室間隔(SP)的質(zhì)量,計(jì)算右心肥厚指數(shù)(RVHI)(右心室和左心室+室間隔的比值)。實(shí)驗(yàn)組,假手術(shù)組的右肺分別進(jìn)行HE染色后做病理切片,分析判斷模型是否符合肺動(dòng)脈高壓的病理生理改變,用免疫組化檢測(cè)TGF-β1在肺動(dòng)脈的表達(dá)情況,qRT-PCR分別檢測(cè)TGF-β1及相關(guān)miRNAs的表達(dá)量。結(jié)果:(1)與假手術(shù)組相比,實(shí)驗(yàn)組的RVSP為49.7±6.82mmhg,RVHI為0.347±0.04,假手術(shù)組的RVSP為20.3±4.53mmhg,RVHI為0.199±0.03(均為P0.01),實(shí)驗(yàn)組較假手術(shù)組RVSP及RVHI均明顯升高。實(shí)驗(yàn)組肺組織切片鏡下可見(jiàn)血管內(nèi)皮層明顯增生,微血管瘤形成,細(xì)小肺血管閉塞、機(jī)化,肺血管平滑肌層和內(nèi)外彈力纖維層均明顯增厚,中膜增厚,血管腔減小,肺動(dòng)脈橫切面積為3級(jí),免疫組化檢測(cè)TGF-β1蛋白表達(dá)顯著高于假手術(shù)組(均為P0.05)。(2)在實(shí)驗(yàn)組動(dòng)物模型肺組織中的TGF-β1的表達(dá)量為假手術(shù)組肺組織的1-3倍,實(shí)驗(yàn)組小鼠肺組織中miRNA-19a表達(dá)較假手術(shù)組增高15-30倍,miRNA-18a表達(dá)增高1-4倍,miRNA-23a表達(dá)1-2.5倍,miRNA-25a表達(dá)較假手術(shù)組增高1-3倍,(P均0.05)。其中實(shí)驗(yàn)組中miRNA-19a表達(dá)較其它miRNAs顯著升高(P0.05)。結(jié)論:左肺切除造成的血流動(dòng)力學(xué)變化可導(dǎo)致小鼠的肺動(dòng)脈高壓形成,這種肺動(dòng)脈高壓的動(dòng)物模型可用于CHD-PAH的實(shí)驗(yàn)室研究。TGF-β1及miRNA-19a的高表達(dá)可能參與了小鼠肺動(dòng)脈高壓模型中肺血管的重塑及肺動(dòng)脈高壓的形成。
[Abstract]:Objective: to investigate the relationship between the expression of TGF- 尾 1 and related mi RNAs and pulmonary vascular remodeling and pulmonary hypertension in mice with dynamic pulmonary hypertension. To investigate whether the expression of TGF- 尾 1 and related mi RNAs is related to the rate of progression of pulmonary hypertension. Methods: the model of pulmonary hypertension in mice was established, and the lung tissue of the model mice was taken as the research object. The effects of hemodynamic changes on the expression of TGF- 尾 1 and related miRNAs in lung tissue were further verified. Methods: 20 male C57 mice were randomly divided into 2 groups: 1. Both groups were anesthetized by 40 mg / kg pentobarbital sodium. The left third intercostal thoracotomy was performed in the experimental group, left lung was excised, and the left third intercostal thoracotomy was performed in the sham operation group. The left lung was not resected. After 8 weeks of operation, the right ventricular systolic pressure (RV SPP), the right ventricular volume (RV) and the left ventricular (LVL) ventricular septum (SPV) were measured in the experimental group and the sham-operated group, respectively. The right ventricular hypertrophy index and the ratio of right ventricle to left ventricular septum were calculated. The expression of TGF- 尾 1 in pulmonary artery was detected by immunohistochemistry. QRT-PCR was used to detect the expression of TGF- 尾 1 and related miRNAs. Results compared with sham operation group, the RVSP of experimental group was 49.7 鹵6.82 mmhg. RVHI was 0.347 鹵0.04, RVSP in sham-operated group was 20.3 鹵4.53mmg / g, RVSP was 0.199 鹵0.03 (all P 0.01). RVSP and RVHI in the experimental group were significantly higher than those in the sham-operated group. In the experimental group, vascular endothelial layer proliferation, microangioma formation, small pulmonary vascular occlusion and organization could be seen in the lung tissue sections of the experimental group. The pulmonary vascular smooth muscle layer and the inner and outer elastic fiber layer were obviously thickened, the middle membrane thickened, the blood lumen decreased, and the transverse area of pulmonary artery was grade 3. The expression of TGF- 尾 1 protein by immunohistochemistry was significantly higher than that in sham-operated group (P 0.05, P < 0.05). The expression of TGF- 尾 1 in lung tissue of experimental group was 1-3 times as much as that of sham operation group. The expression of miRNA-19a in lung tissue of experimental group was 15-30 times higher than that of sham-operated group. The expression of miRNA-18a was 1-4 times higher than that of sham-operated group, and the expression of miRNA-23a was 1-2.5 times higher than that of sham operation group. The expression of miRNA-25a was 1-3 times higher than that of sham operation group. The expression of miRNA-19a in the experimental group was significantly higher than that in other miRNAs groups (P 0.05). Conclusion: hemodynamic changes caused by left lung resection may lead to pulmonary hypertension in mice. The high expression of TGF- 尾 1 and miRNA-19a may be involved in pulmonary vascular remodeling in mice with pulmonary hypertension. And the formation of pulmonary hypertension.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R544.1

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本文編號(hào)：1453806