白细胞介素-1β和黄芪对气管上皮细胞转化生长因子-β_1表达的影响

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。

Effects of IL-1β and astragalus on the expression of transforming growth factor-β1 in cultured airway epithelial cells

Objective To investigate the effects of IL-1β on expression of transforming growth factor-β_1 (TGF-β_1) in cultured airway epithelial cells treated with IL-1β +/- astragalus and the role of astragalus on airway remodeling. Methods The airway epithelial cells cultured ex vivo were randomly divided into control group and treatment group. The treatment groups were treated as follows: ①added with 1ng/ml IL-1β and collected for supernatant and cell-coated dishes after 8, 16, 24 and 48 h; ②added with varied levels of IL-1β(0.1 ng/ml,1 ng/ml and 10 ng/ml); ③added with 10 ng/ml IL-1β and varied levels of astragalus and dexamethasone. For series ② and ③, supernatant and cell-coated dishes were obtained after 24 h. The expression of TGF-β_1 protein in supernatant and cells was measured by double-antibody sandwich ELISA and immunocytochemistry respectively. Results ①The expression of TGF-β_1 in treatment group added with 1 ng/ml IL-1β was significantly higher at 24 h [(0.613±0.022), (701±32)pg/ml] than at other time spots (P<0.05 or P<0.01, respectively); ②Compared with control group [(0.138±0.009), (216±28)pg/ml], the expressions of TGF-β_1 significantly increased in treatment groups added with 0.1 ng/ml IL-1β [(0.156±0.003), (267±12)pg/ml], 1 ng/ml IL-1β [(0.614±0.020), (710±32)pg/ml] or 10 ng/ml IL-1β [(0.917±0.050), (940±34)pg/ml] (P<0.05 or P<0.01, respectively). A positive dose-effect correlation was found between TGF-β_1 level and IL-1β dosage (r=0.906, P<0.01); ③The expressions of TGF-β_1 in groups added with astragalus 50 mg [(0.397±0.020), (398±52)pg/ml], 200 mg [(0.144±0.005), (258±45)pg/ml], 500 mg [(0.401±0.005), (414±22)pg/ml] or dexamethasone [(0.155±0.003),(247±44)pg/ml] were much lower than that in IL-1β_10 ng/ml group [(0.904±0.047), (935±32)pg/ml] (P<0.05 or P<0.01, respectively). Conclusion IL-1β may contribute to the development and progress of airway remodeling by promoted expression of TGF-β_1, an effect which appeared to be inhibited with astragalus.