骨髓间充质干细胞对LPS刺激BV2小胶质细胞炎症因子的影响

Effect of bone marrow mesenchymal stem cells on inflammatory factors released by LPS-stimulated BV-2 microglia

目的:探讨大鼠骨髓间充质干细胞(MSCs)对LPS诱导BV2小胶质细胞炎症反应的影响。方法:采用贴壁分离法分离、纯化大鼠MSCs,BV2小胶质细胞进行常规培养传代。实验分为PBS对照组(A组)、PBS联合MSCs处理组(B组)、LPS刺激组(C组)、LPS联合MSCs干预组(D组)。BV2小胶质细胞接种于6孔板中培养过夜后,按上述实验分组处理,24 h后收集各组培养上清液用于后续实验检测。显微镜下观察BV2小胶质细胞形态变化,Griess法检测NO浓度,ELISA法检测TNF-α、IL-1β的含量。结果:MSCs可以改善活化小胶质细胞的细胞形态;C组中LPS诱导小胶质细胞活化后,培养上清NO、IL-1β、TNF-α的含量明显上升(P<0.05);D组中予MSCs处理后NO、IL-β、TNF-α释放显著减少(P<0.05)。结论:MSCs明显降低活化小胶质细胞炎症因子的释放,其可能是通过抑制小胶质细胞炎症反应而发挥神经保护作用;本研究中MSCs通过非直接接触的方式作用于小胶质细胞,推测没有进入脑内的MSCs对活化小胶质细胞亦有抑制作用。

Objective To explore the effect of bone marrow mesenchymal stem cells （MSCs） on LPS- stimulated BV2 microglia in inflammatory reaction. Methods Mouse MSCs were isolated and purified by adherence screening. The routinely cultured BV2 microglia in vitro were divided into PBS control group （group A）, PBS plus MSCs treatment group （group B）, LPS stimulation group （group C ） and LPS plus MSCs group （group D）.MSCs and BV2 microglia were cultured in the transwell co-culture system for 24 hours. We observed BV2 mieroglia morphological changes under the microscope, detected the concentrations of NO by Griess reaction, and the level of IL-1β,TNF-α by ELISA. Results MSCs can improve the morphology of activated microglia. The concentrations of TNF-α, IL-1β and NO in culture supernatants were increased significantly （P 〈 0.05 ） after microglia activation, however, at the present of MSCs, the concentration of these inflammatory factors declined dramaticly （P 〈 0.05 ）. Conclusions MSCs can significantly inhibit the activation of microglia. It may play a neuroprotective effect by reducing the inflammation of microglia. MSCs showing anti-inflammatory effects through non-direct contact with nieroglial, suggesting that MSCs outside the brain may also inhibit the activation of microglia.