橙皮素早期干预对APPswe/PS1dE9小鼠星形胶质细胞的作用

Effect of Hesperetin Early Intervention on Astroglial APPswe/PS1dE9 in Mice

摘要

目的 研究橙皮素早期干预对野生型及转基因小鼠星形胶质细胞的作用。方法 选取3月龄野生型C57BL/6J小鼠为对照组、对照给药橙皮素10 mg/kg/d、20 mg/kg/d剂量组;设立3月龄转基因小鼠为模型组、橙皮素20 mg/kg/d、40 mg/kg/d、80 mg/kg/d剂量组,灌胃1次/d,灌胃6个月后处死。购买3月龄C57BL/6J小鼠为正常对照组,免疫组化（IHC）检测小鼠脑组织星形胶质细胞活化标志物GFAP表达;免疫荧光双重染色（IF）检测小鼠脑组织GFAP/C3、GFAP/S100A10共定位的细胞表达;蛋白免疫印迹法（WB）检测小鼠脑组织中GFAP、C3的蛋白表达;ELISA法测定小鼠脑组织中IL-1α、TGF-β1含量。结果 与正常对照组相比,对照组GFAP阳性表达上调,GFAP/C3共定位阳性细胞数增加,GFAP/S100A10减少,C3表达上调,IL-1α含量升高。与对照组相比,对照给药橙皮素组GFAP表达下降,GFAP/C3共定位阳性细胞数减少,GFAP/S100A10增加,C3表达下调,IL-1α含量下降,TGF-β1含量升高。与对照组相比,模型组GFAP表达增强,C3蛋白表达上调,GFAP/S100A10阳性细胞数减少,TGF-β1含量下降。与模型组相比,橙皮素给药组GFAP表达下降,GFAP/C3共定位阳性细胞数减少,GFAP/S100A10增加,C3的表达下调,TGF-β1含量升高(P<0.05)。结论 橙皮素早期干预能够明显减轻自然衰老小鼠及转基因小鼠炎症反应,其机制可能与抑制星形胶质细胞活化,促进抗炎A2型细胞活化,抑制促炎A1型活化,减轻炎症对神经元的损伤有关。

Abstract

Objective To explore the effect of hesperetin early intervention to astrocyte in wild type and APPswe/PS1dE9 double transgenic mice. Methods The 3-month-old C57BL/6J wild type mice were set up as control group, hesperetin 10 mg/kg/d and 20 mg/kg/d group. The 3-month-old APPswe/PS1dE9 transgenic mice were set up as model group, hesperetin 20 mg/kg/d, 40 mg/kg/d, and 80 mg/kg/d groups, continuous intragastric administration for 6 months, once a day. The mice were killed after 6 months of intragastric administration, and 3-month-old C57BL/6J mice were purchased as normal control group. Immunohistochemical was used to test the expression of GFAP in mouse brain, double immunofluorescence staining was used to test the co-localized expressions of GFAP/C3 and GFAP/S100A10, Western Blot was used to observe the expression of GFAP and C3 in brain tissues of mice, IL-1α and TGF-β1 in brain tissue of mice were tested by using ELISA. Results Compared with the normal control group, the expression of GFAP and C3 in the control group increased, the number of GFAP/C3 colocalization positive cells increased, while the GFAP/S100A10 decreased. Compared with the control group, hesperetin could decrease the expression of GFAP, the number of GFAP/C3 colocation-positive cells, C3 as well as IL-1α, while the number of GFAP/S100A10 positive cells, TGF-β1 increased. Compared with the control group, the expression of GFAP, C3 in the model group increased, while the number of GFAP/S100A10 positive cells decreased. Compared with the model group, the expression of GFAP, C3 and the number of GFAP/C3 colocation-positive cells in hesperetin group decreased, while the expression of TGF-β1 and the number of GFAP/S100A10 colocation-positive cells increased (P<0.05). Conclusion Hesperetin early intervention can significantly alleviate the inflammation of naturally aged mice and transgenic mice, and its mechanism may be related to inhibiting the activation of astrocytes, promoting the activation of anti-inflammatory A2 cells, inhibiting the activation of pro-inflammatory A1 cells, and reducing the damage of neurons caused by inflammation.

关键词

阿尔茨海默病 / 橙皮素 / 星形胶质细胞 / 早期干预

Key words

Alzheimer's disease / Hesperetin / Astrocyte / Early intervention

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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）