芝麻素调控AMPK/SIRT1自噬通路对阿尔茨海默病模型大鼠学习和记忆能力的影响

doi:10.13481/j.1671-587X.20260309

摘要/Abstract

摘要：

目的探讨芝麻素（Ses）调控单磷酸腺苷（AMP）活化蛋白激酶（AMPK）/沉默信息调节因子1（SIRT1）自噬通路对阿尔茨海默病（AD）大鼠学习和记忆能力的影响。方法 90只雄性SD大鼠随机分为对照组、模型组、低剂量Ses组（Ses-L组）、高剂量Ses组（Ses-H组）、阳性对照多奈哌齐（Donepezil）组和高剂量Ses+AMPK抑制剂Compound C组（Ses-H+Compound C组），每组15只。采用跳台实验检查各组大鼠跳台潜伏期和触电数，Morris水迷宫实验检测各组大鼠逃避潜伏期和穿越平台数，HE染色观察各组大鼠海马CA1区神经元病理形态表现，TUNEL染色检测各组大鼠海马组织中神经元凋亡率，透射电子显微镜观察各组大鼠海马CA1区自噬体的形成情况，Western blotting法检测各组大鼠海马CA1区组织中苄氯素1（Beclin1）、微管相关蛋白1轻链3（LC3）、AMPK、磷酸化AMPK（p-AMPK）和沉默信息调节因子1（SIRT1）蛋白表达水平。结果跳台实验、Morris水迷宫实验和HE染色，与对照组比较，模型组大鼠跳台潜伏期缩短（P<0.05），触电数增加（P<0.05）；逃避潜伏期延长（P<0.05），穿越平台数减少（P<0.05），大部分神经元出现核固缩，正常细胞数目减少，神经元凋亡率升高（P<0.05）。与模型组比较，Ses-L组、Ses-H组和Donepezil组大鼠跳台潜伏期延长（P<0.05），触电数减少（P<0.05）；逃避潜伏期缩短（P<0.05），穿越平台数增加（P<0.05）；海马CA1区神经元结构和形态有所改善，神经元凋亡率降低（P<0.05）。与Ses-L组比较，Ses-H组和Donepezil组大鼠跳台潜伏期延长（P<0.05），触电数减少（P<0.05）；逃避潜伏期缩短（P<0.05），穿越平台数增加（P<0.05），海马CA1区病理损伤进一步减轻，神经元凋亡率降低（P<0.05）。与Ses-H组比较，Ses-H+Compound C组大鼠跳台潜伏期缩短（P<0.05），触电数增加（P<0.05）；逃避潜伏期延长（P<0.05），穿越平台数减少（P<0.05）；海马CA1区可见大量核固缩的神经元，神经元凋亡率升高（P<0.05）。TUNEL染色和Western blotting法，与对照组比较，模型组大鼠海马CA1区自噬体数量减少，Beclin1和SIRT1蛋白表达水平及LC3-Ⅱ/LC3-Ⅰ和p-AMPK/AMPK比值降低（P<0.05）；与模型组比较，Ses-L组、Ses-H组和Donepezil组大鼠海马CA1区自噬体数量增多，Beclin1和SIRT1蛋白表达水平及LC3-Ⅱ/LC3-Ⅰ和p-AMPK/AMPK比值升高（P<0.05）；与Ses-L组比较，Ses-H组和Donepezil组大鼠海马CA1区自噬体数量增多，Beclin1和SIRT1蛋白表达水平及LC3-Ⅱ/LC3-Ⅰ和p-AMPK/AMPK比值升高（P<0.05）；与Ses-H组比较，Ses-H+Compound C组大鼠海马CA1区自噬体数量减少，Beclin1和SIRT1蛋白表达水平及LC3-Ⅱ/LC3-Ⅰ和p-AMPK/AMPK比值降低（P<0.05）。结论 Ses可改善AD模型大鼠学习和记忆能力，可能与Ses激活AMPK/SIRT1自噬通路有关。

关键词: 芝麻素, 腺苷酸活化蛋白激酶/沉默信息调节因子1自噬通路, 阿尔茨海默病, 学习和记忆能力, 自噬

Abstract:

Objective To discuss the effect of sesamin （Ses） regulating adenosine monophosphate （AMP）-activated protein kinase （AMPK）/silent information regulator 1 （SIRT1） autophagy pathway on the learning and memory abilities of rats with Alzheimer’s disease （AD）. Methods Ninety male SD rats were randomly divided into control group， model group， low dose of Ses group （Ses-L group）， high dose of Ses group （Ses-H group）， positive control Donepezil group（Donepezil group）， and high dose of Ses+AMPK inhibitor Compound C group （Ses-H+Compound C group）， with 15 rats in each group. Step-down test was used to examine the step-down latencies and numbers of electric shocks of the rats in various groups； Morris water maze test was used to detect the escape latency and number of platform crossings of the rats in various groups； HE staining was used to observe the pathological morphology of neurons in the hippocampal CA1 region of the rats in various groups； TUNEL staining was used to detect the apoptotic rate of neurons in the hippocampus tissue of the rats in various groups； transmission electron microscope was used to observe the formation of autophagosomes in the hippocampal CA1 region of the rats in various groups； Western blotting method was used to detect the expression levels of rabbit primary antibodies against Beclin1， microtubule-associated protein 1 light chain 3 （LC3）， AMPK， phosphorylated AMPK （p-AMPK）， and SIRT1 proteins in the hippocampus CA1 region tissue of the rats in various groups. Results The step-down test， Morris water maze test and HE staining results showed that compared with control group， the step-down latency of the rats in model group was shortened （P<0.05）， and the number of electric shocks was increased （P<0.05）； the escape latency was prolonged （P<0.05）， and the number of platform crossings was decreased （P<0.05）； most neurons showed pyknosis， the number of normal cells was reduced， and the apoptotic rate of neurons was increased （P<0.05）. Compared with model group， the step-down latencies of the rats in Ses-L group， Ses-H group and Donepezil group were prolonged （P<0.05）， and the numbers of electric shocks were decreased （P<0.05）； the escape latencies were shortened （P<0.05）， and the numbers of platform crossings were increased （P<0.05）； the structure and morphology of neurons in hippocampal CA1 region were improved， and the apoptotic rates of neurons were decreased （P<0.05）. Compared with Ses-L group， the step-down latencies of the rats in Ses-H group and Donepezil group were prolonged （P<0.05）， and the number of electric shocks were decreased （P<0.05）； the escape latency was shortened （P<0.05）， and the numbers of platform crossings were increased （P<0.05）； the pathological damage in hippocampal CA1 region was further alleviated， and the apoptotic rate of neurons were decreased （P<0.05）. Compared with Ses-H group， the step-down latency of the rats in Ses-H+Compound C group was shortened （P<0.05）， and the number of electric shocks was increased （P<0.05）； the escape latency was prolonged （P<0.05）， and the number of platform crossings was decreased （P<0.05）； a large number of pyknotic neurons were observed in hippocampal CA1 region， and the apoptotic rate of neurons was increased （P<0.05）. The TUNEL staining and Western blotting results showed that compared with control group， the number of autophagosomes in hippocampal CA1 region of the rats in model group was decreased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were decreased （P<0.05）； compared with model group， the numbers of autophagosomes in hippocampal CA1 region of the rats in Ses-L group， Ses-H group and Donepezil group were increased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were increased （P<0.05）； compared with Ses-L group， the numbers of autophagosomes in hippocampal CA1 region of the rats in Ses-H group and Donepezil group were increased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were increased （P<0.05）； compared with Ses-H group， the number of autophagosomes in hippocampal CA1 region of the rats in Ses-H+Compound C group was decreased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were decreased （P<0.05）. Conclusion Ses can improve the learning and memory abilities of AD model rats， which may be related to the activation of AMPK/SIRT1 autophagy pathway by Ses.

Key words: Sesamin, Activated protein kinase/Silent information regulator 1 autophagy pathway, Alzheimer’s disease, Learning and memory ability, Autophagy

中图分类号:

R742

王景欣,高月娟,张菁楠,牛佳雯,金春花. 芝麻素调控AMPK/SIRT1自噬通路对阿尔茨海默病模型大鼠学习和记忆能力的影响[J]. 吉林大学学报(医学版), 2026, 52(3): 663-671.

Jingxin WANG,Yuejuan GAO,Jingnan ZHANG,Jiawen NIU,Chunhua JIN. Effect of sesamin on learning and memory abilities of rats with Alzheimer’s disease model through regulating AMPK/SIRT1 autophagy pathway[J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 663-671.

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Group	Step-down latency（t/s）	Number of electrocutions
Control	242.22±18.67	0.75±0.04
Model	115.56±8.73^*	4.26±0.28^*
Ses-L	148.72±10.05^△	3.71±0.24^△
Ses-H	213.35±12.26^△#	1.45±0.12^△#
Donepezil	217.72±11.35^△#	1.49±0.15^△#
Ses-H+Compound C	145.28±8.89^○	3.43±0.22^○

Group	Escape latency（t/s）	Number of crossing platform
Control	10.56±0.82	8.86±0.62
Model	45.23±3.15^*	3.34±0.24^*
Ses-L	36.63±2.27^△	4.95±0.33^△
Ses-H	15.59±1.03^△#	7.39±0.62^△#
Donepezil	14.62±1.17^△#	7.44±0.67^△#
Ses-H+Compound C	32.33±2.73^○	5.25±0.46^○

Group	Apoptotic rate of neurons（η/%）
Control	5.23±0.44
Model	37.75±2.82^*
Ses-L	29.02±2.14^△
Ses-H	11.26±1.02^△#
Donepezil	10.83±1.04^△#
Ses-H+Compound C	24.45±2.03^○

Group	Beclin1	LC3-Ⅱ/LC3-Ⅰ	p-AMPK/AMPK	SIRT1
Control	0.66±0.03	1.18±0.11	0.94±0.07	0.86±0.06
Model	0.21±0.01^*	0.41±0.03^*	0.33±0.02^*	0.39±0.02^*
Ses-L	0.33±0.02^△	0.59±0.04^△	0.49±0.03^△	0.51±0.03^△
Ses-H	0.57±0.04^△#	0.96±0.08^△#	0.85±0.07^△#	0.77±0.06^△#
Donepezil	0.58±0.05^△#	0.98±0.09^△#	0.87±0.06^△#	0.79±0.06^△#
Ses-H+Compound C	0.36±0.02^○	0.64±0.05^○	0.52±0.04^○	0.54±0.03^○