【摘要】：目的：觀察辛伐他汀對(duì)博萊霉素（BLM）誘導(dǎo)肺纖維化大鼠肺組織血管新生及血管內(nèi)皮生長(zhǎng)因子（VEGF）和血小板第4因子（PF4）基因表達(dá)的影響，探討辛伐他汀治療肺纖維化的分子生物學(xué)機(jī)制，為辛伐他汀的臨床應(yīng)用提供理論依據(jù)。方法：6周齡健康SD雄性大鼠96只，采用隨機(jī)數(shù)字表法分成4組，分別為正常對(duì)照組(A組)、博來(lái)霉素組（B組）、醋酸潑尼松治療組(C組)、辛伐他汀治療組(D組)，每組24只。B、C、D組使用博來(lái)霉素溶液以5mg/kg氣管內(nèi)一次性給藥建立肺纖維化的動(dòng)物模型，A組則以同樣的方法氣管內(nèi)一次性注入等體積生理鹽水。C組、D組在博來(lái)霉素造模后第1天開(kāi)始分別以潑尼松混懸液5mg/kg·d、辛伐他汀混懸液10mg/kg·d灌胃，而A組和B組分別以等體積的生理鹽水（10ml/kg）每日灌胃。分別于造模后第7、14、28天取材，各組每次隨機(jī)處死大鼠8只，共32只。肺組織HE染色觀察肺泡炎及肺纖維化改變；采用免疫組化法（SP法）檢測(cè)各組大鼠肺組織血管新生情況及VEGF、PF4蛋白的表達(dá)，采用RT-PCR法檢測(cè)各組大鼠肺組織VEGF及PF4mRNA的表達(dá)。 結(jié)果：①肺組織病理學(xué)結(jié)果顯示A組大鼠肺組織結(jié)構(gòu)清晰完整，無(wú)充血、水腫、滲出，無(wú)纖維化病變。B組第7d表現(xiàn)為輕度肺泡炎，肺泡間隔增寬，較多炎細(xì)胞浸潤(rùn)，可見(jiàn)成纖維細(xì)胞增生；第14d肺泡炎達(dá)到高峰，，肺泡間隔明顯增寬，炎細(xì)胞浸潤(rùn)加重，成纖維細(xì)胞明顯增殖；第28d肺纖維化明顯，肺泡腔塌陷或消失，成纖維細(xì)胞大量增生，可見(jiàn)膠原大量沉積。激素及辛伐他汀治療組肺泡炎和纖維化程度較模型組各相應(yīng)時(shí)間點(diǎn)均有不同程度的減輕。②各組之間相同時(shí)間點(diǎn)比較：B、C組大鼠肺組織微血管密度（MVD）高于A組，差異有統(tǒng)計(jì)學(xué)意義；B、C兩組之間比較，差異無(wú)統(tǒng)計(jì)學(xué)意義；D組微血管密度明顯低于B組，差異有統(tǒng)計(jì)學(xué)意義。③免疫組化和RT-PCR結(jié)果：各組之間相同時(shí)間點(diǎn)比較， B組、C組肺組織VEGF表達(dá)明顯高于A組，差異有統(tǒng)計(jì)學(xué)意義；B、C兩組間比較差異無(wú)統(tǒng)計(jì)學(xué)意義；D組肺組織VEGF表達(dá)明顯低于B組，差異有統(tǒng)計(jì)學(xué)意義。B組、C組肺組織PF4表達(dá)明顯低于A組，差異有統(tǒng)計(jì)學(xué)意義；B、C兩組間比較差異無(wú)統(tǒng)計(jì)學(xué)意義；D組肺組織PF4表達(dá)明顯高于B組，差異有統(tǒng)計(jì)學(xué)意義。結(jié)論：①辛伐他汀能夠抑制肺纖維化大鼠肺組織中的微血管新生，減輕肺泡炎和肺纖維化的程度。②辛伐他汀抗肺纖維化的機(jī)制可能與其抑制肺組織內(nèi)VEGF的表達(dá)，相對(duì)上調(diào)PF4的表達(dá)，抑制病理性血管新生有關(guān)。③糖皮質(zhì)激素不能減輕肺纖維化過(guò)程中肺組織的血管新生。
[Abstract]:Aim: to observe the effects of simvastatin on angiogenesis, vascular endothelial growth factor (VEGF) and platelet factor 4 (PF4) gene expression in lung tissue of bleomycin (BLM) induced pulmonary fibrosis rats. To explore the molecular biological mechanism of simvastatin in the treatment of pulmonary fibrosis and to provide theoretical basis for the clinical application of simvastatin. Methods: 96 healthy SD male rats aged 6 weeks were randomly divided into 4 groups: normal control group (group A), bleomycin group (group B), prednisone acetate treatment group (group C) and simvastatin treatment group (group D). 24 rats in each group were treated with bleomycin solution and intratracheal administration of bleomycin solution to establish pulmonary fibrosis model. Group A was given the same amount of saline in the trachea, and group C was injected with the same volume of saline in the trachea at one time, and the rats in group C were given intratracheal administration of bleomycin in one dose. Group D was treated with prednisone suspension (5mg/kg d) and simvastatin suspension (10mg/kg d) on the first day after bleomycin modeling, while group A and group B were intragastrically perfused with normal saline (10ml/kg) of the same volume. The rats were randomly killed in each group on the 7th day 1428 day after the model, altogether 32 rats were killed. The changes of pulmonary alveolitis and pulmonary fibrosis were observed by HE staining, the expression of VEGF,PF4 protein and angiogenesis in lung tissue were detected by immunohistochemical method (SP), and the expression of VEGF and PF4mRNA in lung tissue were detected by RT-PCR method. Results: 1 the lung histopathological results showed that the lung tissue structure of group A was clear and complete, without hyperemia, edema, exudation and fibrosis. Fibroblast proliferation was observed. On the 14th day, the alveolar inflammation reached the peak, the alveolar septum widened obviously, the inflammatory cell infiltration increased, the fibroblast proliferated obviously, and on the 28th day, the pulmonary fibrosis was obvious, the alveolar cavity collapsed or disappeared, the fibroblasts proliferated and a large amount of collagen was deposited. The pulmonary alveolitis and fibrosis degree of steroid and simvastatin treatment group were alleviated in different degrees compared with the corresponding time points in model group. 2 comparison of the same time points among groups: the (MVD) of lung tissue in group B C was higher than that in group A. The difference was statistically significant. There was no significant difference between the two groups. The microvessel density in group D was significantly lower than that in group B. the difference was statistically significant. 3 Immunohistochemical and RT-PCR results showed that the expression of VEGF in group B and C was significantly higher than that in group A at the same time. The expression of VEGF in group D was significantly lower than that in group B. the expression of PF4 in group B and C was significantly lower than that in group A (P < 0.05). The expression of PF4 in group D was significantly higher than that in group B (P < 0.05). Conclusion: 1 Simvastatin can inhibit the angiogenesis of pulmonary tissue and alleviate the degree of alveolitis and pulmonary fibrosis in rats with pulmonary fibrosis. 2 the mechanism of simvastatin against pulmonary fibrosis may be related to its inhibition of VEGF expression in lung tissue. Relative up-regulation of PF4 expression and inhibition of pathological angiogenesis. 3 glucocorticoid can not reduce pulmonary angiogenesis during pulmonary fibrosis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R563

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

1 管麗麗;吳偉;李平華;劉曉熹;鄢秀菊;;雷帕霉素對(duì)人臍靜脈內(nèi)皮細(xì)胞增殖的影響[J];激光雜志;2010年02期

相關(guān)博士學(xué)位論文 前3條

1 徐延路;生物標(biāo)志物與支架植入術(shù)后冠狀動(dòng)脈粥樣硬化病變進(jìn)展的相關(guān)性研究[D];北京協(xié)和醫(yī)學(xué)院;2011年

2 陳蓓;胃癌血管內(nèi)皮靶向肽GX1在腫瘤生長(zhǎng)及血管生成中的作用及機(jī)制研究[D];第四軍醫(yī)大學(xué);2009年

3 陳良;低剪切力介導(dǎo)內(nèi)皮細(xì)胞表達(dá)膜型基質(zhì)金屬蛋白酶1的機(jī)制及辛伐他汀干預(yù)的研究[D];山東大學(xué);2009年

相關(guān)碩士學(xué)位論文 前4條

1 管麗麗;雷帕霉素抑制兔角膜新生血管的實(shí)驗(yàn)研究[D];重慶醫(yī)科大學(xué);2008年

2 張?jiān)品?多硫酸化磷酸甘露寡糖的制備及其抗腫瘤血管生成作用的研究[D];山東大學(xué);2009年

3 吳春娃;不同培養(yǎng)方式對(duì)人肺癌及臍靜脈內(nèi)皮細(xì)胞基因表達(dá)及差異的影響分析[D];天津醫(yī)科大學(xué);2010年

4 林伊利;三種姜黃色素體外抗腫瘤作用研究[D];浙江中醫(yī)藥大學(xué);2013年

本文編號(hào)：2420530