生慧汤通过调节神经递质改善阿尔茨海默病模型小鼠认知损伤和昼夜节律紊乱

Shenghuitang Mitigates Cognitive Impairment and Circadian Rhythm Disturbance in Mouse Model of Alzheimer's Disease via Regulating Expression Levels of Neurotransmitters

目的:观察生慧汤对阿尔茨海默病(AD)小鼠血清中神经递质含量的影响,并探讨其改善AD认知损伤和昼夜节律紊乱的机制。方法:将27只APP/PS1小鼠随机分为模型组、多奈哌齐组和生慧汤组,另将9只野生型C57BL/6JNju正常小鼠设为空白组。多奈哌齐组(0.92×10^(-4) g·kg^(-1)·d^(-1))和生慧汤组(13.5 g·kg^(-1)·d^(-1))分别灌服多奈哌齐和生慧汤,空白组和模型组灌服等体积的纯水,各组连续灌胃4周。采用Morris水迷宫实验和自主活动实验评估小鼠的认知功能和昼夜节律。采用液相色谱-质谱联用(LC-MS/MS)技术检测小鼠血清中乙酰胆碱(ACh)、胆碱乙酰转移酶(ChAT)、去甲肾上腺素(NE)、肾上腺素(E)、谷氨酸(Glu)、5-羟基吲哚乙酸(5-HIAA)和多巴胺(DA)的表达水平。结果:与空白组比较,模型组小鼠的平台潜伏期、游泳距离、初次抵达平台时间、光照活动时间、黑暗活动时间和总活动时间显著增加(P<0.01),穿越平台次数和目标象限游泳时间则显著减少(P<0.01);与模型组比较,多奈哌齐组和生慧汤组小鼠的平台潜伏期、游泳距离、初次抵达平台时间、光照活动时间、黑暗活动时间和总活动时间减少(P<0.05,P<0.01),穿越平台次数和目标象限游泳时间增加(P<0.05,P<0.01)。与空白组比较,模型组小鼠血清中ACh、ChAT的表达水平显著减少(P<0.01);与模型组比较,多奈哌齐组和生慧汤组小鼠血清中ACh、ChAT的表达水平增加(P<0.05,P<0.01)。与空白组比较,模型组小鼠血清中Glu的表达水平显著增加(P<0.01),NE、5-HIAA和DA的表达水平显著减少(P<0.01);与模型组比较,生慧汤组小鼠血清中Glu的表达水平降低(P<0.05),NE、5-HIAA和DA的表达水平增加(P<0.05,P<0.01);多奈哌齐组小鼠血清中NE、Glu、5-HIAA、DA的表达水平较模型组变化不明显,且差异没有统计学意义。各组小鼠血清中E的表达水平变化不明显,且差异没有统计学意义。结论:生慧汤可改善AD小鼠的认知损伤和昼夜节律紊乱,其机制可能与其调节神经递质有关。

Objective:To observe the effect of Shenghuitang on serum levels of neurotransmitters in the mouse model of Alzheimer's disease(AD)and explore the mechanism of Shenghuitang in mitigating the cognitive impairment and circadian rhythm disturbance of AD.Method:Twenty-seven APP/PS1 dementia mice were randomly assigned into a model group,a donepezil(0.92×10^(-4) g·kg^(-1)·d^(-1))group,and a Shenghuitang(13.5 g·kg^(-1)·d^(-1))group.Another nine wild-type C57BL/6JNju mice was set as the control group.The mice were administrated with corresponding drugs by gavage and the control and model groups were given the same volume of pure water.Every group was continuously treated for 4 weeks.The cognitive function and circadian rhythm of mice were evaluated by Morris water maze test and open field test.The liquid chromatography-tandem mass spectrometry(LC-MS/MS)was employed to determine the expression levels of acetylcholine(ACh),choline acetyltransferase(ChAT),norepinephrine(NE),epinephrine(E),glutamate(Glu),5-hydroxyindoleacetic acid(5-HIAA),and dopamine(DA)in the serum.Result:Compared with the control group,the modeling increased the escape latency,swimming distance,time of first arrival on the platform,activity time of light environment,activity time of dark environment,and total activity time(P<0.01),while it decreased the number of crossing the platform and the swimming time in the target quadrant(P<0.01).Compared with the model group,donepezil and Shenghuitang decreased the escape latency,swimming distance,time of first arrival on the platform,activity time of light environment,activity time of dark environment and total activity time(P<0.05,P<0.01),while they increased the number of crossing the platform and the swimming time in the target quadrant(P<0.05,P<0.01).Compared with the control group,the modeling down-regulated the expression levels of ACh and ChAT in the serum(P<0.01).Compared with the model group,donepezil and Shenghuitang upregulated the expression levels of ACh and ChAT in the serum(P<0.05,P<0.01).Compared with the control group,the modeling up-regulated the expression level of Glu in the serum(P<0.01)and down-regulated the expression levels of NE,5-HIAA,and DA(P<0.01).Compared with the model group,Shenghuitang downregulated the expression level of Glu(P<0.05)and up-regulated the expression levels of NE,5-HIAA,and DA(P<0.05,P<0.01).The expression levels of NE,Glu,5-HIAA,and DA in the donepezil group did not change significantly compared with those in the model group.The expression level of E showed no significant difference among different groups.Conclusion:Shenghuitang may ameliorate the cognitive impairment and circadian rhythm disturbance of AD mice by regulating the levels of neurotransmitters in the serum.