橙皮素早期干预APPswe/PS1d E9小鼠对Aβ转运蛋白的影响

Effect of hesperetin in early intervention on Aβ transporter in APPswe / PS1d E9 mice

摘要

目的 研究不同剂量橙皮素早期干预APPswe/PS1d E9双转基因小鼠对β-淀粉样蛋白（Aβ）相关转运蛋白的影响。方法 三月龄C57BL/6J野生型小鼠为对照组,三月龄APPswe/PS1d E9双转基因小鼠随机分为模型组、橙皮素低、中、高剂量组（20、40、80 mg·kg^-1·d^-1）,每天灌胃1次,连续6个月。免疫组织化学法测定脑组织中胰岛素降解酶（IDE）的表达,蛋白印迹法检测转运Aβ的相关蛋白晚期糖基化终产物（RAGE）、低密度脂蛋白受体相关蛋白1（LRP-1）、P-糖蛋白（P-gp）及血脑屏障（BBB）相关蛋白紧密连接蛋白-5（Claudin-5）、闭合蛋白（Occludin）、闭锁小带蛋白-1（ZO-1）的表达。结果 与模型组相比,橙皮素三个剂量组IDE表达明显增强;与对照组相比,模型组P-gp和LRP-1表达减少,RAGE表达增加,Claudin-5、Occludin、ZO-1表达均减少;与模型组相比,橙皮素干预后P-gp和LRP-1表达增加,RAGE表达减少,Claudin-5、Occludin和ZO-1表达均增加（P<0.05）。结论 橙皮素早期干预APPswe/PS1d E9小鼠可通过增强IDE活性、影响Aβ转运蛋白表达及血脑屏障紧密性,从而影响Aβ代谢和转运。

Abstract

Objective To explore the effect of different doses of hesperetin early intervention on β-amyloid transporter in APPswe/PS1dE9 transgenic mice. Methods The 3-month-old C57BL/6J wild type mice were set up as the control group, the 3-month-old APPswe/PS1d E9 transgenic mice were randomly divided into the model group, hesperetin low-dose group, medium-dose group and high-dose groups (20, 40, 80 mg·kg^-1·d^-1), and gavage once a day for 6 months. Immunohistochemistry was applied to test the expression of insulin-degrading enzyme (IDE) in brain tissue, Western blotting (WB) was applied to detect receptor of advanced glycation endproducts (RAGE), low density lipoprotein receptor-associated protein 1 (LRP-1) , P-glycoprotein (P-gp) and the blood-brain barrier related proteins Claudin-5, Occludin and occlusive zonolin-1 (ZO-1). Results Compared with the model group, IDE expression was significantly enhanced in three hesperetin groups. Compared with the control group, the model group exhibited decreased expressions of P-gp and LRP-1, increased expression of RAGE, and decreased expressions of Claudin-5, Occludin, and ZO-1. In contrast, compared with the model group, the hesperetin groups showed increased expressions of P-gp and LRP-1, decreased expression of RAGE, and increased expressions of Claudin-5, Occludin, and ZO-1(P<0.05). Conclusion Early intervention of hesperetin in APPswe/PS1dE9 mice can affect Aβ metabolismand transport by enhancing IDE activity, regulating Aβ transporter and blood-brain barrier tightness.

关键词

阿尔茨海默病 / 橙皮素 / 血脑屏障 / 胰岛素降解酶 / 晚期糖基化终产物 / P-糖蛋白

Key words

Alzheimer's disease / hesperetin / blood-brain barrier / insulin-degrading enzyme / receptor of advanced glycation endproducts / P-glycoprotein

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基金资助

河北省教育厅高等学校科学技术研究项目（ZD2017002）; 河北省中医药重点学科建设项目（冀中医药函[2021]7号）; 承德医学院神经生物学学科（071006）