TNF-α反义核酸应用于治疗肺纤维化的研究

发布时间：2018-08-30 18:54

【摘要】：肺纤维化,即肺间质组织由胶原蛋白、弹性蛋白及蛋白糖类构成,当纤维母细胞受到化学性或物理性伤害时,会分泌胶原蛋白进行肺间质组织的修补,进而造成肺纤维化。科学家在研究TNF-a在机体内的作用机制时发现TNF-a在肺纤维化的发生发展过程中有着重要的作用,自然而然的TNF-a反义核酸被用于治疗肺纤维化也成了一种新的思路。TNF-a反义核酸用于治疗肺纤维化的潜力巨大,但是全身性的给药TNF-α反义核酸会造成多种负面的结果,比如说感染、免疫缺失以及恶性肿瘤。所以我们尝试通过局部给药TNF-α反义核酸来达到增强其治疗肺纤维化的疗效同时减少副作用的目的。为了研究TNF-a反义核酸在治疗肺纤维化中的作用,我们构建了博莱霉素诱导的C57小鼠肺纤维化模型,在小鼠模型4天、6天、9天、13天的时候,通过鼻腔滴注的方式进行TNF-a反义核酸原位给药,在第14天的时候处死小鼠。通过肺组织切片双标荧光染色的方式检测肺实质组织中的TH2细胞。通过肺部灌洗的方式检测灌洗液中Treg细胞的变化比例；灌洗液上清检测炎症因子TNF-α,IL-13,INF-γ,IL-10,TGF-β的变化水平；通过免疫荧光组织切片的方式检测TNF-a在肺组织的分布；利用组织切片染色,Western blot等检测技术在组织水平对肺纤维化发生发展过程中的各项指标进行分析。通过以上手段,我们在体内和体外条件下研究了TNF-a反义核酸在小鼠肺纤维化中的治疗效果和机理。
[Abstract]:Pulmonary fibrosis, which consists of collagen, elastin and proteoglycoids, will secrete collagen to repair the interstitial tissue of lung, which will lead to pulmonary fibrosis when the fibroblasts are damaged by chemical or physical factors. In studying the mechanism of TNF-a in the body, scientists have found that TNF-a plays an important role in the development of pulmonary fibrosis. Naturally, the use of TNF-a antisense nucleotides in the treatment of pulmonary fibrosis has also become a new way of thinking. TNF-a antisense nucleotides have great potential for treatment of pulmonary fibrosis, but systemic administration of TNF- 伪 antisense nucleotides can lead to a variety of negative results, such as infection. Immune deficiency and malignant tumors. Therefore, we try to enhance the efficacy of TNF- 伪 antisense nucleic acid in the treatment of pulmonary fibrosis and reduce side effects. In order to study the role of TNF-a antisense nucleic acid in the treatment of pulmonary fibrosis, we constructed a bleomycin induced pulmonary fibrosis model in C57 mice. TNF-a antisense nucleic acid was given in situ by nasal drip, and the mice were killed on the 14th day. TH2 cells in lung parenchyma were detected by double-labeled fluorescence staining in lung tissue sections. The percentage of Treg cells in the lavage fluid was detected by the method of lung lavage, the level of the inflammatory factor TNF- 伪, IL-13, INF- 纬, IL-10 and TGF- 尾 was detected by the supernatant of the lavage fluid, and the distribution of TNF-a in the lung tissue was detected by immunofluorescence tissue section. The indexes of the development of pulmonary fibrosis were analyzed at the tissue level by using tissue section staining and Western blot. The therapeutic effect and mechanism of TNF-a antisense oligonucleotides on pulmonary fibrosis in mice were studied in vivo and in vitro.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563

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