辛伐他汀对纤维化大鼠肺组织Snail1、E-钙粘素、波形蛋白表达的影响

发布时间：2018-04-26 20:21

本文选题：肺纤维化 + 辛伐他汀　；参考：《南华大学》2014年硕士论文

【摘要】：【目的】建立博来霉素(bleomycin, BLM)诱导的大鼠肺纤维化模型，探讨Snail1、E-钙粘素（E-cadherin, E-cad）、波形蛋白（Vimentin, VIM）在肺纤维化过程中的表达及辛伐他汀的干预作用。【方法】选用健康雄性SD大鼠60只，随机分为4组，每组15只：A组为对照组；B组为模型组（BLM组）；C组为SIM5mg组，造模后第1天开始连续每天给予辛伐他汀（5mg/kg）溶液灌胃；D组为SIM10mg组，造模后第1天开始连续每天给予辛伐他汀（10mg/Kg）溶液灌胃。B-D组一次性气管内滴注BLM（5mg/Kg）制造肺纤维化动物模型。A组气管内给予等体积的生理盐水。分别于第7、14、28天随机各处死5只，碱性水解法检测肺组织羟脯氨酸（HYP）含量；HE染色观察肺组织肺泡炎和纤维化变化；RT-PCR法检测Snail1mRNA、E-cad mRNA、VIM mRNA的表达；免疫组织化学技术SP法检测Snail1蛋白、E-cad蛋白、VIM蛋白的表达。【结果】 1. HYP含量与A组比较，7d、14d、28d时B、C及D组大鼠肺组织HYP含量均有升高，差异有统计学意义（P㩳0.05），以B组HYP含量最高。与B组比较，C组、D组HYP含量均有下降，D组低于C组，其差异有统计学意义，以D组28d下降明显。 2.病理改变（HE染色） A组大鼠肺组织结构正常，肺泡间隔未见增宽，无炎性细胞浸润，无纤维化改变。B组7d肺组织以炎症反应为主；14d肺组织结构破坏，可见较多炎性细胞和成纤维细胞；28d肺组织结构破坏严重，成纤维细胞增殖，肺纤维化形成。C、D组肺泡炎症及纤维化程度较B组不同程度减轻。 3. Snail1、Vimentin及E-cad表达检测 7d、14d、28d时，A组Snail1、Vimentin为低表达，E-cad为高表达；B组Snail1、Vimentin表达增高，E-cad表达减低，差异有统计学意义（P㩳0.05）；辛伐他汀干预后，与B组比较，C、D组Snail1、Vimentin表达不同程度减低，E-cad表达不同程度升高，其差异均有统计学意义（P0.05），以D组28天最为明显（P0.05）。【结论】 1.辛伐他汀能够减轻BLM诱导的大鼠肺纤维化程度。 2.辛伐他汀抗纤维化机制可能与抑制Snail1、Vimentin表达，促进E-cad表达，，延缓上皮细胞间质转化有关。
[Abstract]:[purpose] A rat model of pulmonary fibrosis induced by bleomycin (BLM) was established to investigate the expression of Snail1 E-cadherin E-cadherin and vimentin Vimentin in the process of pulmonary fibrosis and the intervention of simvastatin. [methods] Sixty male Sprague-Dawley rats were randomly divided into 4 groups. 15 rats in each group were divided into two groups: control group (n = 15), model group B (n = 15), SIM5mg group C (n = 15) and simvastatin group (n = 5 mg / kg). Group D was treated with simvastatin (5 mg / kg) daily as SIM10mg group. One day after the model was established, the rats in group A were given simvastatin 10 mg / kg) solution intravenously intratracheal instillation of BLM5 mg / kg) to make pulmonary fibrosis animal model. Group A was given normal saline in the trachea of the same volume. Five rats were killed randomly on day 7, 14 and 28, respectively. The content of hydroxyproline (HYP) in lung tissue was detected by alkaline hydrolysis method and the changes of pulmonary alveolitis and fibrosis were observed by HE staining. The expression of Snail1 mRNA-E-cad mRNAVIM mRNA was detected by RT-PCR. Immunohistochemistry SP method was used to detect the expression of E-cad protein and vim protein in Snail1. [results] 1. HYP content Compared with group A, the content of HYP in lung tissue of group C and group D increased at 14 days and 28 days, and the difference was statistically significant. The content of HYP in group B was the highest. Compared with group B, the content of HYP in group C was lower than that in group C, and the difference was statistically significant, especially in group D for 28 days. 2. Histopathological changes (HE staining) In group A, the lung tissue structure was normal, alveolar septum was not enlarged, no inflammatory cells were infiltrated, and no fibrosis was observed. It can be seen that more inflammatory cells and fibroblasts destroyed the lung tissue structure seriously at 28 days, the proliferation of fibroblasts and the degree of alveolar inflammation and fibrosis in group C. Con D were reduced compared with those in group B. 3. Detection of vimentin and E-cad expression in Snail1 The expression of Snail1Vimentin in group A was lower than that in group B, and the expression of Vimentin in group B was significantly higher than that in group B (P < 0.05), and the expression of Snail1Vimentin in group C was lower than that in group B, and the expression of E-cad was increased in group C (P < 0.05) after treatment with simvastatin, and the expression of Snail1Vimentin in group A was significantly lower than that in group B (P < 0.05), and the expression of Vimentin in group A was significantly lower than that in group B (P < 0.05). The difference was statistically significant (P 0.05), especially in group D (28 days). [conclusion] 1. Simvastatin can reduce the degree of pulmonary fibrosis induced by BLM in rats. 2. The anti-fibrosis mechanism of simvastatin may be related to the inhibition of Snail1Vimentin expression, the promotion of E-cad expression and the delay of epithelial interstitial transformation.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R563

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