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目的检测经白细胞介素(IL)-1β、IL-1β和黄芪共同干预后兔气道上皮细胞转化生长因子-β1(TGF-β1)蛋白的表达水平,探讨IL-1β对气管上皮细胞TGF-β1表达的影响以及黄芪在哮喘气管重塑中的防治作用。方法体外培养兔气管上皮细胞,①实验组加入终浓度为1ng/ml的IL-1β,于不同时间点收集培养上清液及贴有细胞的盖玻片;②实验组分别加入不同浓度的IL-1β;③各组均加入终浓度为10ng/ml的IL-1β,同时实验组分别加入不同浓度的黄芪和地塞米松。②与③均于24h后收集培养上清液及贴有细胞的盖玻片。采用免疫细胞化学染色和双抗体夹心酶联免疫吸附试验(ELISA)测定TGF-β1蛋白的表达。结果①经IL-1β1ng/ml处理后TGF-β1的表达在24h点[上皮细胞吸光度值(0.613±0.022),上清液含量(701±32)pg/ml]明显高于其余各时间点(P分别<0.05和0.01)。②与对照组[(0.138±0.009),(216±28)pg/ml]比较,IL-1β 0.1ng组[(0.156±0.003),(267±12)pg/ml]、1ng组[(0.614±0.020),(710±32)pg/ml]、10ng组[(0.917±0.050),(940±34)pg/ml]TGF-β1表达均增高,差异有统计学意义(P分别<0.05和0.01);经直线相关分析培养上清液中TGF-β1的含量与所加IL-1β的浓度呈正相关(r=0.906,P<0.01)。③与IL-1β组[(0.904±0.047),(935±32)pg/ml]比较,黄芪50mg组[(0.397±0.020),(398±52)pg/ml]、黄芪200mg组[(0.144±0.005),(258±45)pg/ml]、黄芪500mg组[(0.401±0.005),(414±22)pg/ml]和地塞米松组[(0.155±0.003),(247±44)pg/ml]TGF-β1表达水平均降低,差异有统计学意义(P分别<0.05和0.01)。结论IL-1β可促进气管上皮细胞TGF-β1的蛋白表达,黄芪对这一过程有抑制作用,早期应用黄芪可能延缓气管重塑的形成与发展。
Objective To investigate the expression level of transforming growth factor-β1 (TGF-β1) in airway epithelial cells after intervening with interleukin (IL)-1β, IL-1β and astragalus, and to investigate the effect of IL-1β on the expression of TGF-β1 in tracheal epithelial cells. Effects of β1 Expression and Prevention and Cure of Astragalus in Tracheal Remodeling in Asthma Methods Rabbit tracheal epithelial cells were cultured in vitro. 1 IL-1β was added to the experimental group at a final concentration of 1 ng/ml. Culture supernatants and coverslips with cells were collected at different time points. 2 Different concentrations of IL were added to the experimental groups. IL-1β was added to each group at a final concentration of 10 ng/ml. At the same time, different concentrations of Astragalus and dexamethasone were added to the experimental group. Culture supernatants and cell-covered coverslips were collected after 2 and 3 for 24 h. Immunocytochemical staining and double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) were used to determine the expression of TGF-β1 protein. Results 1 The expression of TGF-β1 after treatment with IL-1β 1 ng/ml was significantly higher than that at the other time points at the 24h point [absorbance of epithelial cells (0.613±0.022), supernatant (701±32) pg/ml] ( P <0.05 and 0.01 respectively). 2 Compared with control group [(0.138±0.009), (216±28) pg/ml], IL-1β 0.1ng group [(0.156±0.003), (267±12) pg/ml], 1ng group [(0.614 The expression of TGF-β1 was increased in ±0.020, (710±32) pg/ml], 10 ng group [(0.917±0.050), (940±34) pg/ml], and the difference was statistically significant (P<0.05 and 0.01); There was a positive correlation between the content of TGF-β1 in culture supernatant and the concentration of IL-1β added by linear correlation analysis (r=0.906, P<0.01). 3Compared with the IL-1β group [(0.904±0.047), (935±32) pg/ml], the Astragalus 50mg group [(0.397±0.020), (398±52) pg/ml] and the Astragalus 200mg group [(0.144 ± 0.005), (258 ± 45) pg/ml], Astragalus 500mg group [(0.401 ± 0.005), (414 ± 22) pg/ml] and dexamethasone group [(0.155 ± 0.003), (247 ± 44) The expression of TGF/β1 in pg/ml] was decreased, and the difference was statistically significant (P<0.05 and 0.01, respectively). Conclusion IL-1β can promote the expression of TGF-β1 in tracheal epithelial cells. Huangqi can inhibit this process. Early application of Huangqi may delay the formation and development of tracheal remodeling.