齐越 李荧 贾冬 杨彩瑜 房晓楠 李纪彤

[摘要] 目的 觀察癫痫清颗粒对AD小鼠模型BBB的影响。 方法 采用侧脑室注射Aβ复制AD模型。选取80只小鼠按体重随机分为假手术组，模型组，癫痫清颗粒12.48 g/kg组、6.24 g/kg组、3.12 g/kg组，盐酸多奈哌齐组，BD1047（sigma-1受体抑制剂）1 mg/kg组，BD1047+癫痫清颗粒12.48 g/kg组，每组各10只。假手术组和模型组给予蒸馏水，各给药组小鼠灌胃相应药物;同时BD1047组和BD1047+癫痫清12.48 g/kg组，则按照1 mg/kg的剂量腹腔注射给药，每日1次，连续给药21 d。末次给药后，Morris水迷宫检测小鼠的学习记忆能力;HE法观察脑组织病理学变化;硫黄素S染色检测脑组织中Aβ表达水平;Western blotting法测定海马和皮质中Occludin、ZO-1、Cx43、PI3K、p-PI3K、AKT、p-AKT蛋白表达水平。 结果 与模型组比较，BD1047+癫痫清颗粒12.48 g/kg组、癫痫清颗粒12.48 g/kg组、6.24 g/kg组及3.12 g/kg组及盐酸多奈哌齐组小鼠穿台次数、p-PI3K及p-AKT蛋白表达水平显著增加，Aβ表达水平降低，occludin、ZO-1及Cx43表达水平均显著升高，其余指标差异无统计学意义（P>0.05）;各给药组小鼠脑组织病理变化均得到不同程度改善。 结论 癫痫清颗粒可激活sigma-1受体，通过PI3K/AKT信号途径，抑制Aβ表达，保护BBB完整性，改善AD学习记忆障碍。

[关键词] 阿尔茨海默病;血脑屏障;Sigma-1受体;小鼠;连接蛋白

[中图分类号] R969.4          [文献标识码] A          [文章编号] 1673-9701（2021）32-0034-06

[Abstract] Objective To observe the effect of Dianxianqing particles on brain blood barrier（BBB） model of mice with Alzheimer′s disease（AD）. Methods The AD model was replicated by lateral ventricular injection of amyloid β-protein （Aβ）. A total of 80 mice were randomly divided into the sham-operated group， the model group， the Dianxianqing particles 12.48 g/kg group， the Dianxianqing particles 6.24 g/kg group， the Dianxianqing particles 3.12 g/kg group， the donepezil hydrochloride group， the BD1047（sigma-1 receptor inhibitor） 1 mg/kg group and the BD1047+Dianxianqing particles 12.48 g/kg group according to their body weight， with 10 mice in each group. The mice in the sham-operated group and the model group were given distilled water and the corresponding drugs were gavaged in each administration group. Meanwhile， the BD1047 group and the BD1047+Dianxianqing particles 12.48 g/kg group were administered intraperitoneally at a dose of 1 mg/kg once daily for 21 days. After the final administration， the learning and memory ability of mice was detected by Morris water maze test. The pathological changes of brain tissues were observed by hematoxylin-eosin staining（HE） method. The expression level of Aβ in brain tissue was detected by thioflavin S staining. The expression levels of occludin， zonula occludens-1（ZO-1）， connexin 43 （Cx43）， phosphatidylinositol 3-kinase （PI3K）， p-PI3K， serine-threonine kinase （AKT） and p-AKT in hippocampus and cortex were determined by Western blotting. Results Compared with the model group， the number of platform crossings and the expression levels of p-PI3K and p-AKT significantly increased in the BD1047+Dianxianqing particles 12.48 g/kg group， the Dianxianqing particles 12.48 g/kg group， the Dianxianqing particles 6.24 g/kg group， the Dianxianqing particles 3.12 g/kg group and the donepezil hydrochloride group. Meanwhile， the expression level of Aβ decreased， and the expression levels of occludin， ZO-1 and Cx43 significantly increased. There were no statistically significant differences in the remaining indicators. The pathological changes in the brain tissues of mice were improved to different degrees in each administration group. Conclusion Dianxianqing particles can activate sigma-1 receptor， as well as inhibit Aβ expression and protect BBB integrity through PI3K/AKT signaling pathway， thus improving learning and memory impairment of mice with AD.

[Key words] Alzheimer′s disease; Blood brain barrier; Sigma-1 receptor; Mice; Connexin

阿尔茨海默病（Alzheimer′s disease，AD）为一种老年人常见的中枢神经系统退行性疾病，临床主要表现为学习记忆能力减退、认知功能障碍，并伴有神经精神异常和行为障碍。AD病因复杂，细胞外淀粉样蛋白（Amyloid，Aβ）沉积被认为是AD的主要病理学特征[1]，并与血脑屏障（Blood-brain barrier，BBB）破坏密切相关[2]。动物和临床研究表明，BBB功能障碍是AD的病理结果，并会加速其疾病进展[3-4]。……