【摘要】：目的：觀察辛伐他汀對博萊霉素（BLM）誘導肺纖維化大鼠肺組織血管新生及血管內(nèi)皮生長因子（VEGF）和血小板第4因子（PF4）基因表達的影響，探討辛伐他汀治療肺纖維化的分子生物學機制，為辛伐他汀的臨床應用提供理論依據(jù)。方法：6周齡健康SD雄性大鼠96只，采用隨機數(shù)字表法分成4組，分別為正常對照組(A組)、博來霉素組（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組使用博來霉素溶液以5mg/kg氣管內(nèi)一次性給藥建立肺纖維化的動物模型，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組在博來霉素造模后第1天開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）檢測各組大鼠肺組織血管新生情況及VEGF、PF4蛋白的表達，采用RT-PCR法檢測各組大鼠肺組織VEGF及PF4mRNA的表達。 結(jié)果：①肺組織病理學結(jié)果顯示A組大鼠肺組織結(jié)構(gòu)清晰完整，無充血、水腫、滲出，無纖維化病變。B組第7d表現(xiàn)為輕度肺泡炎，肺泡間隔增寬，較多炎細胞浸潤，可見成纖維細胞增生；第14d肺泡炎達到高峰，，肺泡間隔明顯增寬，炎細胞浸潤加重，成纖維細胞明顯增殖；第28d肺纖維化明顯，肺泡腔塌陷或消失，成纖維細胞大量增生，可見膠原大量沉積。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時間點均有不同程度的減輕。②各組之間相同時間點比較：B、C組大鼠肺組織微血管密度（MVD）高于A組，差異有統(tǒng)計學意義；B、C兩組之間比較，差異無統(tǒng)計學意義；D組微血管密度明顯低于B組，差異有統(tǒng)計學意義。③免疫組化和RT-PCR結(jié)果：各組之間相同時間點比較， B組、C組肺組織VEGF表達明顯高于A組，差異有統(tǒng)計學意義；B、C兩組間比較差異無統(tǒng)計學意義；D組肺組織VEGF表達明顯低于B組，差異有統(tǒng)計學意義。B組、C組肺組織PF4表達明顯低于A組，差異有統(tǒng)計學意義；B、C兩組間比較差異無統(tǒng)計學意義；D組肺組織PF4表達明顯高于B組，差異有統(tǒng)計學意義。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制可能與其抑制肺組織內(nèi)VEGF的表達，相對上調(diào)PF4的表達，抑制病理性血管新生有關。③糖皮質(zhì)激素不能減輕肺纖維化過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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R563

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本文編號：2420530