本文選題：肺動(dòng)脈高壓 + 野百合堿��； 參考：《江蘇大學(xué)》2017年碩士論文

【摘要】：目的:利用野百合堿(Monocrotaline,MCT)誘導(dǎo)大鼠肺動(dòng)脈高壓(Pulmonary arterial hypertension,PAH)模型,探討曲前列環(huán)素對(duì)PAH的治療作用及分化抑制因子-1(Inhibitor 1of DNA binding,Id1)和分化抑制因子-3(Inhibitor 3of DNA binding,Id3)與PAH之間的關(guān)系。方法:(1)實(shí)驗(yàn)分組:30只大鼠,隨機(jī)分為生理鹽水對(duì)照組、野百合堿模型組和曲前列環(huán)素干預(yù)組,每組10只。模型組和干預(yù)組大鼠給予注射野百合堿誘導(dǎo)其發(fā)生PAH,對(duì)照組以等張等量生理鹽水注射作為對(duì)照。野百合堿注射3周后,干預(yù)組給予曲前列環(huán)素治療2周,對(duì)照組和模型組給予等張等量生理鹽水,實(shí)驗(yàn)5周后觀察各組大鼠一般情況及體質(zhì)量變化以及留取肺組織標(biāo)本以備后續(xù)實(shí)驗(yàn)使用。(2)PAH指標(biāo)測(cè)定:檢測(cè)各組大鼠平均肺動(dòng)脈壓(mean pulmonary arterial pressur,mPAP)、肺動(dòng)脈收縮壓(Pulmonary arterial systolic pressure,PASP)、右心室肥厚指數(shù)(Mean right ventricular pressure,RVHI),經(jīng)HE染色方法觀察肺小動(dòng)脈形態(tài)學(xué)變化,肺小動(dòng)脈管壁面積(WA)占管總面積的百分比(WA%),同時(shí)計(jì)算管壁厚度占動(dòng)脈外徑的百分比(WT%)。(3)肺組織和肺血管中Id1和Id3蛋白的表達(dá)水平測(cè)定:將各組肺組織進(jìn)行病理切片,通過免疫組化檢測(cè)肺組織和肺血管中Id1和Id3蛋白的表達(dá)水平。(4)肺組織中Id1和Id3蛋白及mRNA的表達(dá)水平測(cè)定:免疫印跡技術(shù)和q-PCR技術(shù)檢測(cè)肺組織中Id1和Id3蛋白及其mRNA的表達(dá)水平。(5)統(tǒng)計(jì)分析:實(shí)驗(yàn)所得數(shù)據(jù)用SPSS 17.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析。計(jì)量資料以_x±s表示,多組之間比較使用單因素方差分析,兩兩比較使用LSD-t檢驗(yàn)。P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:(1)模型組和干預(yù)組大鼠與對(duì)照組大鼠比較,活動(dòng)、飲食明顯減少,反應(yīng)較遲鈍,毛色失去光澤;模型組與對(duì)照組比較,體質(zhì)量明顯降低(P0.05);干預(yù)組與模型組相比較,體質(zhì)量與之差異無統(tǒng)計(jì)學(xué)意義(P0.05)。(2)模型組和干預(yù)組大鼠mPAP、PASP、RVHI、WT%、WA%均明顯高于對(duì)照組(P0.05),干預(yù)組大鼠mPAP、PASP、RVHI、WT%、WA%均明顯高于對(duì)照組而明顯低于模型組(P0.05);(3)免疫組化結(jié)果顯示肺組織和肺血管Id1、Id3蛋白在模型組的表達(dá)量明顯低于對(duì)照組(P0.05),而肺組織和肺血管Id1、Id3蛋白在干預(yù)組的表達(dá)量明顯高于模型組(P0.05)。(4)免疫印跡和q-PCR結(jié)果顯示肺組織Id1、Id3蛋白及Id1mRNA、Id3 mRNA在模型組的表達(dá)量明顯低于對(duì)照組(P0.05),而肺組織Id1、Id3蛋白及Id1mRNA、Id3 mRNA在干預(yù)組的表達(dá)量明顯高于模型組(P0.05)。結(jié)論:曲前列環(huán)素對(duì)野百合堿誘導(dǎo)的大鼠PAH有治療作用,其對(duì)肺動(dòng)脈高壓的治療作用與肺組織中Id1、Id3及Id1mRNA、Id3mRNA的表達(dá)量增加有關(guān)。
[Abstract]:Objective: to investigate the effect of triprostacyclin on the treatment of pulmonary arterial hypertensioning (arterial) in rats with pulmonary hypertension induced by monocrotaline (MCT), and to investigate the relationship between the differentiation inhibitor-1 1of DNA binding Id1 and differentiation inhibitor-3 inhibitor or 3of DNA binding Id3) and PAH. Methods the rats were randomly divided into three groups: normal saline control group, monocrotaline model group and triprostacycline intervention group with 10 rats in each group. Rats in the model group and the intervention group were induced to produce PAH by monocrotaline injection, while the control group was treated with isometric saline injection. After 3 weeks of monocrotaline injection, the intervention group was treated with triprostacyclin for 2 weeks, and the control group and the model group were given the same volume of normal saline. After 5 weeks of experiment, we observed the general condition and body mass change of rats in each group, and collected lung tissue samples for further experiment. The mean pulmonary artery pressure (mean pulmonary arterial pressursurmPAPP), pulmonary systolic pressure (PAP) and pulmonary arterial systolic pressure (PAP) were measured in each group, and the pulmonary artery systolic pressure (PAP) and pulmonary arterial systolic pressure (PASP) were measured. The mean right ventricular pressure of right ventricular hypertrophy index (RVHI) was observed by HE staining, and the morphological changes of pulmonary arterioles were observed by HE staining. The area of the pulmonary arteriole wall (WAA) accounted for the percentage of the total area of the vessel, and the percentage of the thickness of the pulmonary wall to the diameter of the artery. WT3) the expression level of Id1 and Id3 protein in the lung tissue and the pulmonary vessels were determined: pathological sections were made in each group of lung tissues. Detection of Id1 and Id3 protein expression level in lung tissue and pulmonary vessels by immunohistochemistry. (4) determination of Id1 and Id3 protein and mRNA expression in lung tissue; Detection of Id1 and Id3 protein and mRNA in lung tissue by Western blotting and q-PCR technique Statistical analysis: the experimental data were analyzed with SPSS 17.0 software. The metrological data were expressed as x 鹵s, single factor analysis of variance (ANOVA) was used in the comparison between multiple groups, and the difference was statistically significant in pairwise comparison using LSD-t test. P05. Results compared with the control group, the rats of the model group and the intervention group were significantly decreased in activity, diet, reaction, loss of luster, body mass of the model group was significantly lower than that of the control group, and that of the intervention group was significantly lower than that of the model group, while that of the model group was significantly lower than that of the control group, and that of the model group was lower than that of the control group. There was no significant difference in body mass between model group and intervention group (P 0.05). The immunohistochemical results showed that lung tissue and pulmonary blood were significantly higher in model group and intervention group than in control group (P 0.05), and in intervention group were significantly higher than those in control group (P < 0.05), but were significantly lower than that in model group (P 0.05 ~ 0. 05 ~ (3) ~ (3) the results of immunohistochemistry showed that the lung tissue and lung blood in the intervention group were significantly higher than those in the control group (P < 0. 05 ~ (0. 05) ~ 0. 05%), and that in the intervention group was significantly higher than that in the control group (P < 0. 05). The expression of Id1 q-PCR protein in the model group was significantly lower than that in the control group (P0.05), while the expression of Id1 + Id3 protein in the lung tissue and pulmonary blood vessel in the intervention group was significantly higher than that in the model group (P0.05). The results of Western blot and q-PCR showed that the expression of Id1 + 3 protein and Id1 mRNAId3 mRNA in the model group was significantly higher than that in the model group. The expression of Id1mRNA-Id3 protein and Id1mRNA-Id3 mRNA in lung tissue in the intervention group was significantly higher than that in the model group. Conclusion: triprostatin has a therapeutic effect on monocrotaline induced PAH in rats. Its therapeutic effect on pulmonary hypertension is related to the increased expression of Id1 PAH 3 and Id1 mRNAs Id3 mRNA in lung tissue.
【學(xué)位授予單位】：江蘇大學(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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