红霉素对博来霉素致大鼠肺纤维化中ILK、E-钙粘素、α-平滑肌蛋白表达的影响

发布时间：2018-05-31 09:46

本文选题：肺纤维化 + 红霉素　；参考：《南华大学》2012年硕士论文

【摘要】：目的使用博来霉素复制大鼠肺纤维化模型，探讨整合素连接激酶（ILK）、E-钙粘素（e-cadherin，E-cad）、α-平滑肌蛋白（α-SMA）在肺纤维化过程中的表达及红霉素的干预作用。方法健康雄性SD大鼠90只，随机分为6组，每组15只，分别为第1组正常对照组；第2组模型组；第3组造模前三天连续给红霉素，然后予以博莱霉素（bleomycin，BLM）造模；第4组BLM造模后2h开始给红霉素，共给14d；第5组BLM造模后第14d开始给红霉素，至第28d；第6组BLM造模前三天连续给红霉素至造模后第28d。2至6组气管内予以BLM(5mg/g)制作肺纤维化动物模型，，1组气管内予以等体积生理盐水，造模后各组按方案进行给药。分别于7、14、28d随即各处死5只，取左肺组织放于4%甲醛固定后做HE染色及免疫组化检测ILK、E-cad、α-SMA蛋白的表达；取右肺组织进行羟脯氨酸检测和RT-PCR检测ILKmRNA、E－cadherin mRNA、α-SMA mRNA的表达。结果 1.HE染色显示：第2组7d组织以炎症反应为主，可见肺成纤维细胞；14d见大量炎性细胞浸润，较多肺成纤维细胞；28d见大量肺成纤维细胞，说明肺纤维化模型形成。第2至6组病理改变趋势相同，逐渐向肺纤维化发展，但第6组纤维化程度最轻。 2.HYP含量：7d、14d、28d与空白对照组比，2-6组含量均增高，且P＜0.05，以第2组升高最为明显；3、5组各时间段HYP检测与2组相比无明显变化；4、6组予以红霉素干预后，含量均有所下降，且P＜0.05以第6组28d最为显著，P＜0.01。 3.RT-PCR、免疫组化显示：在正常肺组织中，ILK、α-SMA表达为低表达，E-cad为高表达。肺纤维化发生时，与正常组织相比较，ILK、α-SMA表达增强（P＜0.05），E-cad表达减弱（P＜0.05）；予以红霉素干预后，与模型组相比较，第4、6组ILK、α-SMA表达水平下降，E-cad上升（P＜0.05），以第6组28d最为明显（P＜0.01）. 结论1、红霉素可以预防博来霉素所致大鼠肺纤维化，其效果与早期用药及疗程有关。2、红霉素防治肺纤维化作用机制可能与抑制ILK、α-SMA表达，增高E-cad水平，延缓上皮细胞-间质转化发展过程有关。
[Abstract]:Purpose To investigate the expression of E-cadherine-cadherin (E-cadherin) and 伪 -smooth muscle protein (伪 -SMA) in pulmonary fibrosis induced by bleomycin in rats, and to investigate the effect of erythromycin on it. Method 90 healthy male Sprague-Dawley rats were randomly divided into 6 groups, each group (n = 15): normal control group (group 1), model group (group 2), erythromycin (Erythromycin) (group 3) and bleomycin (BLM) (group 3). In group 4, erythromycin was given 2 hours after BLM was made, and erythromycin was given to BLM in group 5 on the 14th day. On the 28th day, the rats in the sixth group were given erythromycin three days before the model was made, and the rats in the 28d.2 to group 6 were given BLMN 5mg / g intratracheal administration. The rats in group 1 were given the same volume of normal saline in trachea. After the model was made, each group was given drugs according to the plan. The left lung tissues were fixed with 4% formaldehyde for HE staining and immunohistochemistry to detect the expression of ILK E-cadE and 伪 -SMA protein, and the right lung tissues were detected for hydroxyproline and RT-PCR for the expression of IL-K mRNA-E-cadherin mRNAin and 伪 -SMA mRNA. Result 1.HE staining showed that in group 2, inflammatory reaction was dominant at day 7. A large number of inflammatory cells were observed in lung fibroblasts on day 14, and a large number of fibroblasts were found in more fibroblasts on day 28, indicating the formation of pulmonary fibrosis model. The pathological changes in groups 2 to 6 showed the same trend and gradually developed to pulmonary fibrosis, but the degree of fibrosis in group 6 was the least. The content of 2.HYP in the control group was significantly higher than that in the control group for 14 days and 28 days, and the content of HYP in the second group was significantly higher than that in the control group (P < 0.05). The content of HYP in the second group was significantly higher than that in the control group (P < 0.05). And P < 0.05 was the most significant in the 6th group on the 28th day (P < 0.01). 3. RT-PCR.Immunohistochemistry showed that the expression of 伪 -SMA was low and the expression of 伪 -SMA was high in normal lung tissues. Compared with the normal tissue, the expression of ILK, 伪 -SMA and E-cad were decreased (P < 0.05) and the expression of 伪 -SMA was decreased (P < 0.01) in group 4 (P < 0.05), compared with that in model group (P < 0.05) after the intervention of erythromycin, the expression level of 伪 -SMA was decreased (P < 0.05) in group 4 (P < 0.05), especially in group 6 (P < 0.01) on the 28th day after treatment with Erythromycin, the expression of 伪 -SMA decreased significantly (P < 0.05), and the expression of 伪 -SMA increased significantly (P < 0.05). Conclusion 1.Erythromycin can prevent pulmonary fibrosis induced by bleomycin in rats, and its effect is related to early medication and course of treatment. The mechanism of erythromycin on pulmonary fibrosis may be to inhibit the expression of ILK, 伪 -SMA and increase E-cad water. It is related to delaying the process of epithelial-mesenchymal transformation.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.9;R965

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