曲前列环素对肺动脉高压大鼠肺组织分化抑制因子表达的影响

发布时间：2018-05-01 06:01

本文选题：肺动脉高压 + 野百合碱　；参考：《江苏大学》2017年硕士论文

【摘要】：目的:利用野百合碱(Monocrotaline,MCT)诱导大鼠肺动脉高压(Pulmonary arterial hypertension,PAH)模型,探讨曲前列环素对PAH的治疗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)与PAH之间的关系。方法:(1)实验分组:30只大鼠,随机分为生理盐水对照组、野百合碱模型组和曲前列环素干预组,每组10只。模型组和干预组大鼠给予注射野百合碱诱导其发生PAH,对照组以等张等量生理盐水注射作为对照。野百合碱注射3周后,干预组给予曲前列环素治疗2周,对照组和模型组给予等张等量生理盐水,实验5周后观察各组大鼠一般情况及体质量变化以及留取肺组织标本以备后续实验使用。(2)PAH指标测定:检测各组大鼠平均肺动脉压(mean pulmonary arterial pressur,mPAP)、肺动脉收缩压(Pulmonary arterial systolic pressure,PASP)、右心室肥厚指数(Mean right ventricular pressure,RVHI),经HE染色方法观察肺小动脉形态学变化,肺小动脉管壁面积(WA)占管总面积的百分比(WA%),同时计算管壁厚度占动脉外径的百分比(WT%)。(3)肺组织和肺血管中Id1和Id3蛋白的表达水平测定:将各组肺组织进行病理切片,通过免疫组化检测肺组织和肺血管中Id1和Id3蛋白的表达水平。(4)肺组织中Id1和Id3蛋白及mRNA的表达水平测定:免疫印迹技术和q-PCR技术检测肺组织中Id1和Id3蛋白及其mRNA的表达水平。(5)统计分析:实验所得数据用SPSS 17.0统计软件进行数据分析。计量资料以_x±s表示,多组之间比较使用单因素方差分析,两两比较使用LSD-t检验。P0.05为差异有统计学意义。结果:(1)模型组和干预组大鼠与对照组大鼠比较,活动、饮食明显减少,反应较迟钝,毛色失去光泽;模型组与对照组比较,体质量明显降低(P0.05);干预组与模型组相比较,体质量与之差异无统计学意义(P0.05)。(2)模型组和干预组大鼠mPAP、PASP、RVHI、WT%、WA%均明显高于对照组(P0.05),干预组大鼠mPAP、PASP、RVHI、WT%、WA%均明显高于对照组而明显低于模型组(P0.05);(3)免疫组化结果显示肺组织和肺血管Id1、Id3蛋白在模型组的表达量明显低于对照组(P0.05),而肺组织和肺血管Id1、Id3蛋白在干预组的表达量明显高于模型组(P0.05)。(4)免疫印迹和q-PCR结果显示肺组织Id1、Id3蛋白及Id1mRNA、Id3 mRNA在模型组的表达量明显低于对照组(P0.05),而肺组织Id1、Id3蛋白及Id1mRNA、Id3 mRNA在干预组的表达量明显高于模型组(P0.05)。结论:曲前列环素对野百合碱诱导的大鼠PAH有治疗作用,其对肺动脉高压的治疗作用与肺组织中Id1、Id3及Id1mRNA、Id3mRNA的表达量增加有关。
[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 卤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.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R544.1

【相似文献】