[1]郭珂一,周峥宇,赵幸娟.葛根素对阿尔茨海默病果蝇的神经保护作用及机制[J].新乡医学院学报,2022,39(6):513-518.[doi:10.7683/xxyxyxb.2022.06.003]
　GUO Keyi,ZHOU Zhengyu,ZHAO Xingjuan.Neuroprotective effect and mechanism of puerarin on drosophila with Alzheimer′s disease[J].Journal of Xinxiang Medical University,2022,39(6):513-518.[doi:10.7683/xxyxyxb.2022.06.003]
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葛根素对阿尔茨海默病果蝇的神经保护作用及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
39
期数:
2022年6
页码:
513-518
栏目:
基础研究
出版日期:
2022-06-05

文章信息/Info

Title:
Neuroprotective effect and mechanism of puerarin on drosophila with Alzheimer′s disease
作者:
郭珂一1; 周峥宇2; 赵幸娟2
（1.郑州大学附属郑州中心医院神经内科，河南　郑州　450000；2.郑州大学第五附属医院神经内科，河南　郑州　450000）
Author(s):
GUO Keyi1; ZHOU Zhengyu2; ZHAO Xingjuan2
(1.Department of Neurology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,Henan Province,China;2.Department of Neurology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,Henan Province,China)
关键词:
阿尔茨海默病; Aβ转基因果蝇; 葛根素; β-淀粉样蛋白
Keywords:
Alzheimer′s disease; Aβ transgenic drosophila; puerarin; amyloid β-protein
分类号:
R741
DOI:
10.7683/xxyxyxb.2022.06.003
文献标志码:
A
摘要:
目的　探讨葛根素对阿尔茨海默病（AD）模型果蝇的神经保护作用及机制。方法　将p〔Gal4〕A307品系的果蝇置于恒温培养箱中培养，收集羽化6 h内的雌蝇为p〔Gal4〕A307处女蝇；将p〔Gal4〕A307处女蝇分别与UAS-Aβ42arc雄蝇杂交，收集子一代果蝇标记为Aβarc果蝇；将p〔Gal4〕A307处女蝇与W1118雄蝇杂交，收集子一代果蝇为正常对照果蝇。取Aβarc果蝇随机分为AD模型组、0.10 mmol·L-1葛根素干预组、0.20 mmol·L-1葛根素干预组、0.24 mmol·L-1葛根素干预组，0.10 mmol·L-1葛根素干预组、0.20 mmol·L-1葛根素干预、0.24 mmol·L-1葛根素干预组果蝇分别用终浓度为0.10、0.20、0.24 mmol·L-1葛根素干预，AD模型组及正常对照组果蝇用含24 g·L-1酵母的超纯水干预。于干预第21天，应用爬管实验检测各组果蝇爬行高度；根据果蝇爬行高度，确定0.20 mmol·L-1为葛根素最适浓度，记录正常对照组、AD模型组、0.20 mmol·L-1葛根素干预组果蝇死亡情况，应用酶联免疫吸附法测定3组果蝇脑组织中Aβ相对表达量，免疫荧光法检测3组果蝇脑组织中Aβ斑块面积。结果　AD 模型组、0.10 mmol·L-1葛根素干预组、0.20 mmol·L-1葛根素干预组、0.24 mmol·L-1葛根素干预组果蝇爬管高度显著低于正常对照组，0.10 mmol·L-1葛根素干预组、0.20 mmol·L-1葛根素干预组、0.24 mmol·L-1葛根素干预组果蝇爬管高度显著高于AD模型组，0.20 mmol·L-1葛根素干预组果蝇爬管高度显著高于0.10 mmol·L-1葛根素干预组和0.24 mmol·L-1葛根素干预组（P<0.05）；0.10 mmol·L-1葛根素干预组与0.24 mmol·L-1葛根素干预组果蝇爬管高度比较差异无统计学意义（P>0.05）。AD模型组、0.20 mmol·L-1葛根素干预组果蝇的中位生存期显著短于正常对照组，0.20 mmol·L-1葛根素干预组果蝇的中位生存期显著长于AD模型组（P<0.05）。AD模型组、0.20 mmol·L-1葛根素干预组果蝇脑组织中Aβ表达水平显著高于正常对照组，0.20 mmol·L-1葛根素干预组果蝇脑组织中Aβ表达水平显著低于AD模型组（P<0.05）。AD模型组、0.20 mmol·L-1葛根素干预组果蝇脑组织中Aβ斑块面积占比显著高于正常对照组，0.20 mmol·L-1葛根素干预组果蝇脑组织中Aβ斑块面积占比显著低于AD模型组（P<0.05）。结论　葛根素可通过降低脑组织内Aβ水平、减少Aβ沉积对AD果蝇发挥神经保护作用，进而改善AD果蝇的爬行能力，延长其寿命。
Abstract:
Objective　To investigate the neuroprotective effect and mechanism of puerarin on drosophila with Alzheimer′s disease (AD).Methods　The p〔Gal4〕 A307 strain drosophila were cultured in a constant temperature incubator.The female drosophila within 6 hours of emergence were collected as p〔Gal4〕 A307 virgin drosophila.The p〔Gal4〕 A307 virgin drosophila were hybridized with UAS-Aβ42arc male drosophila,and the first generational drosophila were labeled as Aβarc drosophila.The p〔Gal4〕 A307 female drosophila were hybridized with W1118 male drosophila,and the first generational drosophila was collected as the normal control drosophila.The Aβarc drosophila were randomly divided into AD model group,0.10 mmol·L-1 puerarin intervention group,0.20 mmol·L-1 puerarin intervention group,and 0.24 mmol·L-1 puerarin intervention group.The drosophila in the 0.10 mmol·L-1 puerarin intervention group,0.20 mmol·L-1 puerarin intervention group and 0.24 mmol·L-1 puerarin intervention group were treated with 0.1,0.2 and 0.24 mmol·L-1 puerarin,respectively.The drosophila in the AD model group and normal control group were treated with ultrapure water containing 24 g·L-1 yeast.On the 21st day of intervention,the climbing height of drosophila in each group was detected by tube climbing experiment.According to the climbing height of drosophila,the 0.20 mmol·L-1 was determined as the optimal concentration of puerarin;the survival days of drosophila in the normal control group,AD model group and 0.20 mmol·L-1 puerarin intervention group were recorded;the expression level of Aβ in brain tissues of drosophila in the three groups was determined by enzyme-linked immunosorbent assay,and the area of Aβ plaque in brain tissue of drosophila in the three groups was determined by immunofluorescence.Results　The climbing height of drosophila in the AD model group,0.10 mmol·L-1 puerarin intervention group,0.20 mmol·L-1 puerarin intervention group and 0.24 mmol·L-1 puerarin intervention group was significantly lower than that in the normal control group(P<0.05).The climbing height of drosophila in the 0.10 mmol·L-1 puerarin intervention group,0.20 mmol·L-1 puerarin intervention group and 0.24 mmol·L-1 puerarin intervention group was significantly higher than that in the AD model group(P<0.05).The climbing height of drosophila in the 0.20 mmol·L-1 puerarin group was significantly higher than that in the 0.10 mmol·L-1 puerarin group and 0.24 mmol·L-1 puerarin group (P<0.05).There was no significant difference in the climbing height of drosophila between the 0.10 mmol·L-1 puerarin group and 0.24 mmol·L-1 puerarin group (P>0.05).The median survival time of drosophila in the AD model group and 0.20 mmol·L-1 puerarin intervention group was significantly shorter than that in the normal control group,and the median survival time of drosophila in the 0.20 mmol·L-1 puerarin intervention group was significantly longer than that in the AD model group (P<0.05).The expression level of Aβ in brain tissue of drosophila in the AD model group and 0.20 mmol·L-1 puerarin intervention group was significantly higher than that in the normal control group,and the expression level of Aβ in brain tissue of drosophila in the 0.20 mmol·L-1 puerarin intervention group was significantly lower than that in the AD model group (P<0.05).The ratio of area of Aβ plaques in brain tissue of drosophila in the AD model group and 0.20 mmol·L-1 puerarin intervention group was significantly higher than that in normal control group,and the ratio of area of Aβ plaques in brain tissue of drosophila in the 0.20 mmol·L-1 puerarin intervention group was significantly lower than that in the AD model group (P<0.05).Conclusion　Puerarin can play a neuroprotective role for drosophila with AD by reducing Aβ level and inhibiting Aβ deposition in brain tissue,which can improve the crawling ability and prolong the life of AD drosophila.

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更新日期/Last Update: 2022-06-05