吉非替尼通过增加Foxo3a活性抑制肺纤维化小鼠上皮-间质转分化

发布时间：2018-08-05 09:31

【摘要】：目的:研究吉非替尼对肺纤维化小鼠转录因子叉头框蛋白O3a(Foxo3a)活性、α-平滑肌肌动蛋白(α-SMA)水平及相关通路的影响,探讨吉非替尼抑制肺上皮-间质转分化的可能机制。方法:将30只SPF级雌性昆明小鼠随机分为3组:对照组(生理盐水气管内雾化)、博来霉素组(博来霉素3 mg/kg溶于100μL生理盐水气管内雾化)和吉非替尼处理组(博来霉素气管内雾化后,每天吉非替尼20 mg/kg溶于100μL生理盐水灌胃)。实验第14天收集样本,将小鼠肺组织置于10%中性甲醛固定,石蜡包埋切片后行HE与Masson染色;RT-PCR法检测Foxo3a和α-SMA mRNA表达水平;Western blotting法检测表皮生长因子受体(EGFR)、Akt、Foxo3a和α-SMA蛋白表达水平。结果:吉非替尼处理组小鼠肺组织病理损伤较博来霉素组明显减轻,胶原沉积明显减少,炎症损伤评分及纤维化评分明显下降(均P0.01),Foxo3a mRNA表达水平明显升高(P0.05),α-SMA mRNA表达水平明显下降(P0.05),总Foxo3a蛋白表达增加,但Foxo3a磷酸化水平显著下降(P0.01),胞核Foxo3a蛋白明显增加(P0.05);同时,EGFR和Akt磷酸化水平也显著下降(P0.01,P0.05),上皮-间质转分化标志蛋白α-SMA表达水平明显降低(P0.05)。结论:吉非替尼抑制博来霉素诱导的肺纤维化,其机制可能与抑制EGFR/Akt通路活化、增强转录因子Foxo3a活性、从而抑制上皮-间质转分化密切相关。
[Abstract]:Aim: to study the effects of gefitinib on the activity of transcriptional factor fork box protein O3a (Foxo3a), the level of 伪 -smooth muscle actin (伪 -SMA) and its related pathways in pulmonary fibrosis mice, and to explore the possible mechanism of gifitinib inhibiting lung epithelial-interstitial transdifferentiation. Methods: thirty SPF female Kunming mice were randomly divided into three groups: control group (normal saline atomization), bleomycin group (bleomycin 3 mg/kg dissolved in 100 渭 L saline atomization) and gefitinib treatment group (Bofetini treatment group). After nebulizing in the trachea, Gefitinib was dissolved in 100 渭 L saline for 20 mg/kg. Samples were collected on the 14th day of the experiment. The lung tissues of mice were fixed with 10% neutral formaldehyde. The expression levels of Foxo3a and 伪 -SMA mRNA were detected by HE and Masson staining after paraffin embedded sections. The expression levels of (EGFR) AktAktto3a and 伪 -SMA proteins were detected by Western blotting. Results: compared with bleomycin group, the pathological injury of lung tissue in the group treated with gefitinib was significantly alleviated and the deposition of collagen was significantly decreased. The expression of Foxo3a mRNA was significantly increased (P0.05), the expression of 伪 -SMA mRNA was significantly decreased (P0.05), and the expression of total Foxo3a protein was increased. However, the level of Foxo3a phosphorylation was significantly decreased (P0.01), the level of Foxo3a protein in nucleus increased significantly (P0.05), and the phosphorylation level of Akt and EGFR decreased significantly (P0.01P 0.05), and the expression of 伪 -SMA, the marker of epithelial-interstitial differentiation, decreased significantly (P0.05). Conclusion: gefitinib inhibits bleomycin-induced pulmonary fibrosis by inhibiting the activation of EGFR/Akt pathway, enhancing the activity of transcription factor Foxo3a, and thus inhibiting the epithelial-interstitial transdifferentiation.
【作者单位】：南方医科大学研究生学院;广州军区广州总医院呼吸内科;
【基金】：广东省自然科学基金资助项目(No.S2011010000511) 广东省科技计划项目(No.2010B031600122) 广东省自然科学基金资助项目(No.9151001002000014) 吴阶平医学基金会临床科研专项资助基金资助项目(No.320.6750.12330)
【分类号】：R563.9

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