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。
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 H37 Rv. 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 PI3 K, 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 H37 Rv. 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. PI3 K 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, PI3 K 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. PI3 K 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 PI3 K/AKT signaling pathway, which leads to the excessive immune response of type II alveolar cells infected by Mtb and the development of inflammation.
作者
苏维娜
李晓晶
张丽丽
包志伟
SU Weina;LI Xiaojing;ZHANG Lili;BAO Zhiwei(Clinical Laboratory,Shandong Normal University Hospital,Jinan 250014,China;Shandong Academy of Traditional Chinese Medicine,Jinan 250014;Department of Clinical Laboratory,Yishui People’s Hospital,Linyi 276400;Department of Internal Medicine,Jianghan University Hospital,Wuhan 430056)
出处
《中国比较医学杂志》
CAS
北大核心
2021年第11期48-54,61,共8页
Chinese Journal of Comparative Medicine
基金
山东省重点研发计划项目(2018GSF119019)。
