麻黄碱对脂多糖诱导的小胶质细胞功能损伤的修复作用及机制

Repair effect of ephedrine on lipopolysaccharide-induced microglia function injury and its mechanism

目的研究麻黄碱对脂多糖(LPS)诱导的小胶质细胞功能损伤的修复作用及机制。方法以人小胶质细胞HMC3为研究对象,考察不同质量浓度麻黄碱(75、150、300、600μg/mL)对HMC3细胞活力和凋亡的影响。然后将HMC3细胞分为对照组(不受药物干预)、LPS组(1μg/mL)、麻黄碱组(1μg/mL LPS+300μg/mL麻黄碱)、BAY11-7082组[1μg/mL LPS+5μmol/L核因子κB(NF-κB)信号通路抑制剂BAY11-7082]、抑制剂组(1μg/mL LPS+300μg/mL麻黄碱+5μmol/L BAY11-7082)和激活剂组(1μg/mL LPS+300μg/mL麻黄碱+1μmol/L NF-κB信号通路激活剂Prostratin),加入相应药物作用24 h后,检测细胞迁移能力和细胞中可溶性白细胞介素6(sIL-6)、白细胞介素10(IL-10)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平以及NF-κB信号通路相关蛋白表达水平。结果300μg/mL的麻黄碱可使HMC3细胞活力显著升高、凋亡率显著降低(P<0.05)。与对照组相比,LPS组迁移细胞数显著增多,细胞中sIL-6、MDA水平和NF-κB蛋白磷酸化水平均显著升高,IL-10、SOD水平均显著降低(P<0.05)。与LPS组相比,麻黄碱组和BAY11-7082组上述指标水平均显著逆转(P<0.05)。与麻黄碱组相比,抑制剂组迁移细胞数显著减少,细胞中sIL-6、MDA水平和NF-κB蛋白磷酸化水平均显著降低,IL-10、SOD水平均显著升高(P<0.05);而激活剂组上述指标水平均显著逆转(P<0.05)。结论麻黄碱可通过抑制LPS诱导的HMC3细胞凋亡、迁移、炎症和氧化应激,修复细胞功能损伤,其作用机制可能与抑制NF-κB信号通路活性有关。

OBJECTIVE To study the repair effect of ephedrine on lipopolysaccharide(LPS)-induced microglia function injury and its mechanism.METHODS Human microglia cells(HMC3)were used as research objects to investigate the effects of different concentrations of ephedrine(75,150,300,600μg/mL)on the viability and apoptosis of HMC3 cells.HMC3 cells were divided into control group(without drug intervention),LPS group(1μg/mL),ephedrine group(1μg/mL LPS+300μg/mL ephedrine),BAY11-7082 group[1μg/mL LPS+5μmol/L nuclear factor-κB(NF-κB)pathway inhibitor BAY11-7082],inhibitor group(1μg/mL LPS+300μg/mL ephedrine+5μmol/L BAY11-7082)and activator group(1μg/mL LPS+300μg/mL ephedrine+1μmol/L NF-κB pathway activator Prostratin).After 24 hours of drug treatment,cell migration,the levels of soluble interleukin-6(sIL-6),interleukin-10(IL-10),superoxide dismutase(SOD)and malondialdehyde(MDA),and the expressions of NF-κB pathway-related proteins were all detected.RESULTS The viability of HMC3 cells could be increased significantly by 300μg/mL ephedrine,while the apoptotic rate was decreased significantly(P<0.05).Compared with the control group,the number of migrating cells was increased significantly in the LPS group;the levels of sIL-6 and MDA,the phosphorylation of NF-κB protein were increased significantly,while the levels of IL-10 and SOD were decreased significantly(P<0.05).Compared with the LPS group,the above indexes were reversed significantly in the ephedrine group and BAY11-7082 group(P<0.05).Compared with the ephedrine group,the number of migrating cells was decreased significantly in the inhibitor group;the levels of sIL-6 and MDA,the phosphorylation of NF-κB protein were decreased significantly,while the levels of IL-10 and SOD were increased significantly(P<0.05).The above indexes were reversed significantly in the activator group(P<0.05).CONCLUSIONS Ephedrine can repair cell injury by inhibiting LPS induced apoptosis,migration,inflammation and oxidant stress of HMC3 cells,the mechanism of which may be associated with inhibiting the activity of the NF-κB signaling pathway.