甘露特纳胶囊改善阿尔兹海默病认知障碍并降低神经炎症的机制研究

The mechanism of GV-971 improving cognitive impairment and reducing neuroinflammation in Alzheimer′s disease

目的探究口服甘露特钠(Sodium Oligomannate Capsules,GV-971)对阿尔兹海默病(Alzheimer′s disease,AD)小鼠认知及神经炎症的影响机制。方法购买雌性ADAPP/PS1小鼠,通过口服GV-971对AD小鼠进行治疗。采用Morris水迷宫实验及Y迷宫实验检测各组小鼠的认知功能,硫黄素S染色检测Aβ(β-淀粉样蛋白)沉积,酶联免疫吸附测定试剂盒(enzyme-linked immunosorbent assay,ELISA)检测各组小鼠大脑中炎性细胞因子1β(interleukin-1β,IL-1β),肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和IL-10的水平。实时定量聚合酶链式反应(quantitative real time polymerase chain reaction,qRT-PCR)及Western blot分别检测各组小鼠脑组织中BACE1 mRNA及蛋白的表达水平。结果健康组和对照组的逃避潜伏期随时间的推移均呈缩短趋势,健康组比对照组趋势更为明显,治疗组的逃避潜伏期随时间的推移呈现先缩短后延长后缩短的趋势,3组组间、时点间以及组间·时点间交互作用差异有统计学意义(P<0.05),治疗组在目标象限停留的时间百分比少于健康组和对照组(P<0.05)。治疗组通过Y迷宫的准确率少于健康组和对照组(P<0.05)。与健康组相比,对照组大脑皮层和海马区Aβ斑块占比和斑块面积升高(P<0.05)。与对照组相比,治疗组小鼠大脑皮层和海马区Aβ斑块占比和Aβ斑块面积均降低(P<0.05)。ELISA结果显示,与健康组相比,对照组小鼠脑组织炎性细胞因子IL-1β和TNF-α水平显著增高,而IL-10水平降低(P<0.05)。与对照组相比,治疗组IL-1β和TNF-α水平均降低,而IL-10水平上升(P<0.05)。与健康组相比,对照组小鼠脑组织中BACE1的mRNA和蛋白水平均升高(P<0.05)。与对照组相比,治疗组小鼠大脑中BACE1的mRNA和蛋白表达下降(P<0.05)。结论GV-971降低脑组织BACE1的表达,改善AD小鼠认知障碍并且降低其神经炎症。

Objective To investigate the mechanism of the effect of oral administration of sodium oligomannate capsules(GV-971)on cognition and neuroinflammation in Alzheimer′s disease(AD)mice.Methods Female APP/PS1 mice with Alzheimer′s disease were purchased and treated with oral GV-971.The cognitive function of mice was tested by Morris water maze test and Y-maze test.Amyloidβ-protein(Aβ)deposition was detected by thioflavin S staining,and the levels of inflammatory cytokines interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and IL-10 in the brains of each group of mice were measured by enzyme-linked immunosorbent assay(ELISA)kits.Quantitative real time polymerase chain reaction(qRT-PCR)and Western blot were used to detect the expression levels of BACE1 mRNA and protein in the brain tissue of mice,respectively.Results The escape latency of the healthy group and the control group showed a decreasing trend over time,with the healthy group showing a more significant trend than the control group.The escape latency of the treatment group showed a trend of initially shortening,then prolonging,and then shortening over time.There was a significant difference in the interaction between groups,time points,and time points between groups(P<0.05).The percentage of time spent in the target quadrant in the treatment group was less than that in the healthy group and the control group(P<0.05).The accuracy of Y-maze passage in the treatment group was lower than that in the healthy group and control group(P<0.05).Compared with the healthy group,the percentage and area of Aβplaques in cerebral cortex and hippocampus the control group were increased(P<0.05),which,however,were decreased in the treatment group,as compared with the control group,(P<0.05).The ELISA results showed that compared with the healthy group,inflammatory cytokines IL-1βand TNF-αlevels in brain tissue of mice in the control group mice were increased,while the IL-10 levels were decreased(P<0.05).Compared with the control group,IL-1βand TNF-αlevels in the treatment group were decreased,while the IL-10 levels were increased(P<0.05).Compared with the healthy group,the mRNA and protein levels of BACE1 in the brain tissue of mice in the control group were increased(P<0.05).Compared with the control group,the mRNA and protein expression of BACE1 in the brain of mice in the treatment group was decreased(P<0.05).Conclusion GV-971 decreased the expression of BACE1 in the brain tissue,improved cognitive impairment and reduced neuroinflammation in AD mice.