交泰丸通过调控AMPK信号通路改善阿尔茨海默病小鼠相关脂代谢异常

doi:10.12122/j.issn.1673-4254.2026.10.19

摘要/Abstract

摘要：

目的探讨交泰丸对阿尔茨海默病（AD）脂代谢调控作用的AMP活化蛋白激酶（AMPK）信号通路机制。方法 20只3月龄雄性APP/PS1双转基因小鼠，随机分为模型组、交泰丸组，10只C57BL/6J野生型雄性小鼠为正常对照组。经4周灌胃干预后取样，通过油红O染色评估各组小鼠脑内脂质沉积情况，生化试剂盒测定血浆及脑组织中总胆固醇（TC）、甘油三酯（TG）和游离脂肪酸（NEFA）水平。Western blotting技术检测脑组织中β分泌酶1（BACE1）、β淀粉样蛋白沉积（Aβ）、磷酸化Tau（p-Tau）、紧密连接蛋白（ZO-1）、闭合蛋白（Occludin）、AMP激活蛋白激酶（AMPK）、磷酸化AMPK（p-AMPK）、固醇调节元件结合蛋白1（SREBP1）、乙酰辅酶A羧化酶（ACC）、脂肪甘油三酯脂肪酶（ATGL）及载脂蛋白E（APOE）的蛋白表达水平。细胞实验中，将成熟脂肪细胞与PC12细胞共培养并分为正常组、模型组、交泰丸干预组、AICAR组、交泰丸联合AICAR组。通过油红O染色观察PC12细胞内脂滴沉积现象，生化试剂盒测定培养基上清液和细胞内TG及NEFA含量。免疫荧光技术评估细胞内AMPK活性，Western blotting检测AMPK、p-AMPK及ACC蛋白表达水平。结果动物实验表明，与模型组比较，交泰丸组小鼠脑内脂质沉积减少；血浆内TC、TG水平下调（P<0.01），NEFA浓度升高（P<0.05）；脑组织内的TG水平降低（P<0.01），NEFA含量降低（P<0.05），而TC未见明显变化。此外，脑内淀粉样蛋白Aβ表达下调（P<0.01），p-Tau表达降低（P<0.05），Occludin表达上调（P<0.01）；代谢通路相关指标显示，交泰丸组小鼠脑内p-AMPK/AMPK总蛋白比值升高（P<0.01），ACC表达降低（P<0.01），SREBP1及APOE表达表达降低（P<0.01，P<0.05），而ATGL无显著性差异。细胞实验结果显示，与模型组相比，交泰丸组细胞内脂质蓄积明显减少；细胞中TG、NEFA水平分别降低（P<0.05，P<0.01），细胞培养上清中TG浓度下降（P<0.01），NEFA释放量增加（P<0.05）；细胞内Aβ水平降低（P<0.01），AMPK活性增强，p-AMPK/AMPK比值升高（P<0.05），ACC表达降低（P<0.01），与AICAR联用有协同增效作用，p-AMPK/AMPK比值升高（P<0.01）。结论交泰丸可以激活AMPK活性，调节下游脂代谢蛋白表达以及脂代谢产物，从而改善AD中脂质沉积和脂代谢紊乱的现象，且对血脑屏障和神经细胞具有保护作用。

关键词: 阿尔茨海默病, 交泰丸, 脂代谢, AMPK信号通路

Abstract:

Objective To explore whether Jiaotaiwan ameliorates Alzheimer disease (AD) by regulating lipid metabolism via the AMPK pathway. Methods Twenty 3-month-old male transgenic APP/PS1 mice were randomized equally into two groups for treatment with Jiaotaiwan or normal saline via gavage for 4 weeks, with 10 saline-treated wild-type C57BL/6J mice as the control group. Brain lipid deposition, plasma and brain TC, TG and NEFA levels, and expressions of BACE1, Aβ, p-Tau, ZO-1, occludin, AMPK/p-AMPK, SREBP1, ACC, ATGL, and APOE in the brain tissues were detected. In the in vitro experiment, PC12 cell co-cultured with mature adipocytes were treated with Jiaotaiwan, AICAR, or both, and the intracellular lipid droplets, TG, NEFA, AMPK activity, and ACC expression were measured. Results In the AD mouse models, Jiaotaiwan significantly reduced brain lipid deposits, lowered plasma TC and TG levels, increased plasma NEFA level, and decreased brain TG and NEFA expressions without affecting TC expression. Jiaotaiwan treatment also significantly lowered the expressions of cerebral Aβ and p-Tau, increased cerebral occludin expression and p-AMPK/AMPK ratio, and reduced ACC, SREBP1 and APOE expressions without affecting ATGL expression in the mouse models. In the co-cultured cells, Jiaotaiwan obviously alleviated cellular lipid accumulation, lowered cellular TG and NEFA levels, reduced supernatant TG content, and enhanced NEFA release. In both the AD mouse model and co-cultured cell model, Jiaotaiwan significantly lowered Aβ level, increased AMPK activity and p-AMPK/AMPK expression, and lowered the expressions of ACC. Jiaotaiwan combined with AICAR produced a synergistic effect for enhancing p-AMPK/AMPK expressions in the co-cultured cells. Conclusion In the APP/PS1 mouse model of AD, Jiaotaiwan alleviates AD-related lipid deposition and dysregulation by activating AMPK and modulating the downstream lipogenic effectors, providing also protective effects on the blood-brain barrier and neural cells.

Key words: Alzheimer disease, Jiaotaiwan, lipid metabolism, AMPK signaling pathway

阮毓卿, 崔璨, 李菁哲, 王妍, 王秀. 交泰丸通过调控AMPK信号通路改善阿尔茨海默病小鼠相关脂代谢异常[J]. 南方医科大学学报, 2026, 46(10): 2509-2517.

Yuqing RUAN, Can CUI, Jingzhe LI, Yan WANG, Xiu WANG. Jiaotaiwan improves lipid metabolism abnormalities in a mouse model of Alzheimer disease by regulating the AMPK signaling pathway[J]. Journal of Southern Medical University, 2026, 46(10): 2509-2517.

图/表 7

图1 交泰丸对APP/PS1双转基因小鼠Aβ蛋白及血脑屏障的影响

Fig.1 Effect of Jiaotaiwan on Aβ protein and blood-brain barrier in APP/PS1 mice. *P<0.05, **P<0.01 vs APP/PS1 group.

图2 交泰丸对APP/PS1双转基因小鼠血脂的影响

Fig.2 Effect of Jiaotaiwan on lipid metabolism in APP/PS1 double transgenic mice. *P<0.05, **P<0.01 vs APP/PS1 group. TC: Total cholesterol; TG: Triglyceride; NEFA: Non-esterified fatty acid.

图3 脑组织油红O染色

Fig.3 Oil Red O staining of the brain tissues of the mice (Upper panel: original magnification： ×20; lower panel: ×200).

图4 交泰丸对APP/PS1双转基因小鼠脑脂的影响

Fig.4 Effect of Jiaotaiwan on brain lipids in AD mouse models. *P<0.05, **P<0.01 vs APP/PS1 group.

图5 交泰丸对AD模型细胞脂质沉积的影响

Fig.5 Effect of Jiaotaiwan on lipid deposition in AD mouse models (Scale bar=200 μm).

图6 交泰丸对AD模型细胞内外脂代谢产物的影响

Fig.6 Effects of Jiaotaiwan on intracellular and extracellular lipid metabolic products in PC12 cell model of AD. A: Intracellular TG and NEFA content. B: TG and NEFA content in the culture supernatant. *P<0.05, **P<0.01 vs Aβ25-35 group.

图7 交泰丸对AMPK/ACC通路的影响

Fig.7 Effect of Jiaotaiwan on the AMPK/ACC pathway. A: Effect of Jiaotaiwan on the AMPK pathway in APP/PS1 double transgenic mice. B: Effects of Jiaotaiwan on the AMPK/ACC pathway in AD model cells. C: Effect of Jiaotaiwan on AMPK activity in AD model cells (Scale bar=25 μm). D: Effect of Jiaotaiwan-medicated serum combined with the AMPK agonist on AD model cells. *P<0.05, **P<0.01 vs Aβ25-35 group.

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