滋肾醒脑汤通过TLR4/NF-κB/NLRP3抑制APP/PS1小鼠小胶质细胞焦亡改善认知功能研究*

作者：邱华安1，2，胡国恒3，李宇翔1，曾 阳3，谢 丹3，王 焱3，盛 望3

单位：1.湖南中医药大学，湖南 长沙 410208； 2.浏阳市中医医院，湖南 浏阳 410300； 3.湖南中医药大学第一附属医院，湖南 长沙 410007

引用：引用：邱华安，胡国恒，李宇翔，曾阳，谢丹，王焱，盛望.滋肾醒脑汤通过TLR4/NF-κB/NLRP3抑制APP/PS1小鼠小胶质细胞焦亡改善认知功能研究[J].中医药导报,2025,31(7):25-31.

DOI：10.13862/j.cn43-1446/r.2025.07.004

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摘要：

目的：探讨滋肾醒脑汤对APP/PS1小鼠小胶质细胞焦亡及认知功能的影响。方法：将50只APP/PS1小鼠随机分为模型组、滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组，每组10只。另取10只C57BL/6J小鼠为正常组。各组分别灌胃给予相应药物。采用水迷宫行为学评定各组小鼠学习及记忆能力；苏木精-伊红（HE）染色观察小鼠脑组织形态；尼氏染色观察小鼠神经元损伤；透射电镜观察小胶质细胞形态；免疫组化分析海马及皮层β-淀粉样蛋白1~42（Aβ1~42）沉积情况；蛋白免疫印记法（Western blotting）法检测各组海马中Toll样受体4（TLR4）、核因子κB（NF-κB）、核苷酸结合寡聚结构域样受体蛋白3（NLRP3）和胱天蛋白酶-1（Caspase-1）蛋白表达情况。结果：模型组、滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠逃避潜伏期均长于正常组（P<0.01或P<0.05）；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠逃避潜伏期均短于模型组（P＜0.05）。模型组小鼠平台象限滞留时间短于正常组（P＜0.05），且跨越平台次数少于正常组（P＜0.05）；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠平台象限滞留时间均长于模型组（P＜0.05或P＜0.01）；滋肾醒脑汤中剂量组及多奈哌齐组小鼠穿越平台次数多于模型组（P＜0.01）。HE染色显示正常组小鼠海马区CA1区神经元排列整齐，形态正常；模型组小鼠海马区CA1区神经元结构不完整，受损的神经元数量增多；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠海马区CA1区小鼠神经元结构、数量及形态均有所改善，仅有少数神经元核空泡丢失，偶见异质细胞。尼氏染色显示正常组小鼠神经元内尼氏体分布均匀，细胞形态规则、排列紧密；模型组小鼠神经元存活数量少、排列稀疏，神经元明显肿胀且胞内尼氏体分布稀疏；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠海马组织中的损伤神经元显著减少，神经元数量、形态及结构均明显改善。免疫组化染色显示正常组小鼠海马CA1区、皮质区神经元形态规则；模型组较正常组细胞形态异常，神经元周围存在明显的深褐色Aβ1~42蛋白沉积；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠Aβ1~42蛋白沉积明显改善，细胞受损情况明显改善。透射电镜下正常组小鼠海马小胶质细胞形态结构正常；模型组小鼠海马神经元明显变性坏死，可见大量呈空泡状的焦亡小体；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠神经元皱缩情况明显改善，胞膜较为完整。模型组小鼠海马组织TLR4、NF-κB、NLRP3及Caspase-1蛋白相对表达量高于正常组（P<0.01）；滋肾醒脑汤低剂量组、滋肾醒脑汤中剂量组、滋肾醒脑汤高剂量组及多奈哌齐组小鼠海马组织TLR4、NF-κB、NLRP3及Caspase-1蛋白相对表达量均低于模型组（P<0.01）。结论：滋肾醒脑汤可能通过抑制TLR4/NF-κB/NLRP3信号通路活性，降低TLR4、NF-κB、NLRP3及Caspase-1的表达，抑制APP/PS1小鼠小胶质细胞焦亡，减少Aβ1~42沉积，发挥神经保护作用，改善认知功能。

关键词：阿尔茨海默病；滋肾醒脑汤；TLR4/NF-κB/NLRP3通路；小胶质细胞；细胞焦亡；认知功能；小鼠

Abstract：

Objective: To investigate the effects of Zishen Xingnao decoction (ZSXND) on microglial pyroptosis and cognitive function in APP/PS1 mice. Method: Totally 50 APP/PS1 mice were randomly divided into model group, ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group, 10 mice in each group, and 10 C57BL/6J mice were selected as the normal group. Drugs were administered via gavage. Learning and memory abilities were assessed using the Morris water maze test. Hematoxylin-eosin (HE) staining was used to observe neuronal morphology and Nissl staining was employed to evaluate neuronal damage. Transmission electron microscopy (TEM) was used to observe microglial ultrastructure. Immunohistochemistry was used to analyze Aβ1~42 deposition in the hippocampus and cortex. Western blotting quantified hippocampal protein levels of toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), NOD-like receptor protein3 (NLRP3) and Caspase-1. Results: Model group, ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group showed longer escape latency than normal group (P<0.01 or P<0.05). ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group exhibited shorter escape latency than model group (P<0.05). The model group showed reduced platform quadrant retention time than normal group (P<0.05) and model group showed fewer platform crossings than normal group (P<0.05). ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group showed longer platform retention time than model group (P<0.05 or P<0.01). ZSXND medium-dose group and donepezil group showed increased platform crossings than model group (P<0.01). HE staining showed that the neurons in the CA1 region of the hippocampus were arranged neatly and had normal morphology in normal group. There were disorganized and damaged neurons in the hippocampal CA1 region of the model group, whereas ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group showed improved neuronal structure, count, and morphology, with only minimal nuclear vacuolization and occasional heterochromatin. Nissl staining showed that Nissl bodies were evenly distributed in neurons in normal group, with regular cell morphology and tight arrangement. There were significantly reduced Nissl bodies, sparse arrangement, and cellular swelling in model group neurons. These damages were markedly ameliorated in ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group. Immunohistochemical staining showed regular morphology of neurons in the hippocampal CA1 and cortical regions in normal group. There were prominent brownish Aβ1~42 deposits surrounding neurons with abnormal morphology in model group. Aβ1~42 deposition and neuronal damage were significantly reduced in ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group. According to TEM, the morphology and structure of hippocampal microglia were normal in normal group. There were degenerated neurons and numerous vacuolated pyroptotic bodies in model group, while ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group showed improved cell shrinkage and relatively intact cell membranes. Model group showed higher TLR4, NF-κB, NLRP3, and Caspase-1 expression in hippocampal than normal group (P<0.01). ZSXND low-dose group, ZSXND medium-dose group, ZSXND high-dose group, and donepezil group showed lower TLR4, NF-κB, NLRP3, and Caspase-1 expression in hippocampal than model group (P<0.01). Conclusion: Zishen Xingnao decoction may exert neuroprotective effects and improve cognitive function by inhibiting the TLR4/NF-κB/NLRP3 signaling pathway, reducing the expression of TLR4, NF-κB, NLRP3, and Caspase-1, inhibiting microglial pyroptosis, and mitigating Aβ1~42 deposition.

Key words：Alzheimer disease; Zishen Xingnao decoction; TLR4/NF-κB/NLRP3 signaling pathway; microglia; cell pyroptosis; cognitive function; mouse

发布时间：2026-01-06

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