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苏维娜,李晓晶,张丽丽,包志伟.miR-214 调控 FGF9 的表达参与结核分枝杆菌感染肺泡Ⅱ型细胞中 PI3K/ AKT 信号通路的免疫调控作用研究[J].中国比较医学杂志,2021,31(11):48~54,61.
miR-214 调控 FGF9 的表达参与结核分枝杆菌感染肺泡Ⅱ型细胞中 PI3K/ AKT 信号通路的免疫调控作用研究
Immunoregulatory mechanism of miR-214 in Mycobacterium tuberculosis infection of alveolar type II cells
投稿时间:2020-12-16  
DOI:10. 3969 / j.issn.1671-7856. 2021. 11. 008
中文关键词:  miR-214  FGF9  PI3K/ AKT 信号通路  结核分枝杆菌  肺泡Ⅱ型细胞  免疫调控
英文关键词:miR-214  FGF9  PI3K/ AKT pathway  Mycobacterium tuberculosis  alveolar type II cells  immunization
基金项目:
作者单位E-mail
苏维娜 山东师范大学校医院检验科,济南 250014 bingtang2000@ 126.com 
李晓晶 山东中医药研究院,济南 250014  
张丽丽 山东省临沂市沂水人民医院检验科, 山东 临沂 276400  
包志伟 江汉大学医院内科,武汉 430056 908925713@ qq.com 
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中文摘要:
       目的 探究 miR-214 对结核分枝杆菌感染肺泡Ⅱ型细胞中免疫调控的作用机制。 方法 培养肺泡 Ⅱ型细胞系 A549,分别使用结核分枝杆菌疫苗株 BCG 和结核分枝杆菌标准株 H37Rv 感染 A549 细胞。 对 A549 细胞进行分组,并进行 H37Rv 感染。 qRT-PCR 检测各组细胞中 miR-214 和 FGF9 的表达情况,Western blot 检测各组细胞中 FGF9 蛋白的表达。 ELISA 检测细胞中免疫相关因子 SP-A、SP-D、TLR2 和 TLR4,炎症因子 IL-10、TNF-α、 TNF-γ、IL-1β 和 IL-8 的含量。 Western blot 检测 PI3K/ AKT 信号通路成员 PI3K、AKT、p-AKT 的表达。 结果 和 BCG 相比,H37Rv 感染后的 A549 细胞中 miR-214 表达升高,FGF9 表达降低。 和 NC mimic+oe-NC 组相比,miR-214 mimic+oe-NC 组细胞 SP-A、SP-D、IL-10 含量显著降低,TLR2、TLR4、TNF-α、TNF-γ、IL-1β、IL-8 含量显著升高,PI3K、 AKT/ p-AKT 表达显著升高。 NC mimic+oe-FGF9 组 SP-A、SP-D、IL-10 含量显著升高,TLR2、TLR4、TNF-α、TNF-γ、IL- 1β、IL-8 含量显著降低,PI3K、AKT/ p-AKT 表达显著升高降低。 和 miR-214 mimic+oe-NC 组相比,miR-214 mimic+ oe-FGF9 组 SP-A、SP-D、IL-10 含量显著升高,TLR2、TLR4、TNF-α、TNF-γ、IL-1β、IL-8 含量显著降低,PI3K、AKT/ p- AKT 表达显著降低。 结论 miR-214 通过抑制 FGF9 的表达,进而激活 PI3K/ AKT 信号通路,导致结核分枝杆菌感染肺泡Ⅱ型细胞过度免疫反应,从而导致炎症发展。
英文摘要:
       Objective To explore the mechanism of miR-214 in immunoregulation of type II alveolar cells infected by Mycobacterium tuberculosis. Methods A549 cells were infected with BCG and H37Rv. qRT-PCR was used to measure the expression levels of miR-214 and FGF9. Western blot was used to measure the expression level of FGF9. SP- A, SP-D, TLR2, TLR4, IL-10, TNF-α, TNF-γ, IL-1β and IL-8 were measure by ELISA. Western blot was used to detect expression of PI3K, AKT and p-AKT. Results Compared with those in cells infected with BCG, expression of miR-214 was increased, and expression of FGF9 was decreased in A549 cells infected with H37Rv. Compared with those in the NC- mimic+oe-NC group, SP-A, SP-D and IL-10 were decreased significantly, TLR2, TLR4, TNF-α, TNF-γ, IL-1β and IL-8 were increased significantly. PI3K and AKT/ p-AKT expression was increased significantly in the miR-214-mimic+oe-NC group, SP-A, SP-D and IL-10 were increased significantly. TLR2, TLR4, TNFα, TNF-γ, IL-1 β and IL-8 were decreased significantly, PI3K and AKT/ p-AKT expression was decreased significantly in the NC mimic+oe-FGF9 group. Compared with those in the NC-mimic+oe-NC group, SP-A, SP-D and IL-10 were increased significantly, TLR2, TLR4, TNF-α, TNF-γ, IL-1 β and IL-8 were decreased significantly. PI3K and AKT/ p-AKT expression was decreased significantly in the miR-214 mimic+oe-FGF9 group. Conclusions miR-214 inhibits FGF9 expression and activates the PI3K/ AKT signaling pathway, which leads to the excessive immune response of type II alveolar cells infected by Mtb and the development of inflammation.
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