摘要
目的:研究人参皂苷Re对瞬时转染瑞典型突变淀粉样前体蛋白(APPswe)cDNA的小鼠神经瘤母细胞(N2a细胞)内β2-肾上腺受体(β2-AR)及β-样淀粉蛋白肽(Aβ)的影响。方法:瞬时转染APPswe质粒入N2a细胞24h后,加入1、10及100μmol/L浓度的人参皂苷Re处理细胞24h后收集细胞。应用点印迹检测4G8(识别Aβ17-24片段)的表达,免疫印迹检测APP、β2-AR、β2-AR丝氨酸346位点磷酸化(pS346-β2-AR)和β2-AR苏氨酸68位点磷酸化(pT68-β2-AR)的表达,并采用免疫细胞化学技术分析统计pS346-β2-ARβ及β2-AR的染色强度。结果:瞬时转染APPswe质粒的N2a细胞内pS346-β2-AR的表达较pcDNA组增强(P<0.05),pT68-β2-AR的表达较pcDNA组减弱(P<0.05)。与APPswe组比较,1、10、100μmol/L的人参皂苷Re可降低pS346-β2-AR的表达(P<0.01),人参皂苷Re不能逆转pT68-β2-AR的表达水平。免疫细胞化学统计显示,转染APPswe质粒的N2a细胞内pS346-β2-AR染色强度较对照组显著增强(P<0.05),人参皂苷Re可减弱其表达。β2-AR的表达在各组无显著差异。转染APPswe质粒的N2a细胞内APP及4G8的表达较对照组显著增加(P<0.01);与APPswe组比较,比较,100μmol/L的人参皂苷Re可减少APP蛋白的表达(P<0.05),10μmol/L及100μmol/L的人参皂苷Re降低4G8表达(P<0.05)。结论:人参皂苷Re可降低瞬时转染APPswe质粒的N2a细胞内pS346-β2-AR的表达水平,并显著减少该细胞内APP及Aβ的表达,改善AD的基本病变。
Objective:To explore the effects of Ginsenoside Re onβ2-adrenergic receptor(β2-AR)andβ-amyloid(Aβ)in transiently transfected Swedish amyloid precursor proteins-695 mutation plasmid(APPswe)Neuro2 a(N2 a)cells.Methods:N2 a cells were transiently transfected with APPswe cDNA,then the cells were treated with Ginsenoside Re at a concentration of1,10 and 100μmol/L for 24 h.Western Blot was used to detect APP expression.The expression of 4 G8(recognition of Aβ17-24 fragment)was detected by Dot blot.For measurement ofβ2-AR,Immunocytochemistry and Western Blot were used to analyze totalβ2-AR,phosphorylation ofβ2-AR at serine 346(pS346-β2-AR)and threonine at 68(pT68-β2-AR).Results:pS346-β2-AR was increased(P<0.05),while pT68-β2-AR was decreased in transiently transfected APPswe N2 a cells compared with control group(P<0.05).Ginsenoside Re with the concentration of 1,10 and 100μmol/L reduced pS346-β2-AR(P<0.01),however Ginsenoside Re could not reverse the expression of pT68-β2-AR.Immunocytochemistry results showed that staining intensity of pS346-β2-AR increased in transiently transfected APPswe N2 a cells compared with control group(P<0.05),Ginsenoside Re could weaken its expression.There were no significant differences of totalβ2-AR in 5 groups.APP and 4 G8 expression in transiently transfected APPswe N2 a cells increased significantly compared with control group(P<0.01),while compared with model group,100μmol/L Ginsenoside Re could decrease the expression of APP(P<0.01),yet 10μmol/L and 100μmol/L Ginsenoside Re could decrease the expression of 4 G8(P<0.05).Conclusion:Ginsenoside Re can decreases pS346-β2-AR,APP and Aβexpression in transiently transfected APPswe N2 a cells,and may improve Alzheimer’s disease basic lesions.
作者
姜霞
刘萍
李凡
王平
JIANG Xia;LIU Ping;LI Fan;WANG Ping(Department of Pathology,Hubei University of Chinese Medicine,Wuhan430065,China;Institute of Gerontology,Hubei University of Chinese Medicine,Wuhan430065,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第6期2397-2401,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81303252)
湖北省自然科学基金项目(No.2017CFB733)
第九批中国博士后基金特别资助(No.2016T90685)~~
作者简介
通讯作者:王平,湖北省武汉市洪山区黄家湖西路1号湖北中医药大学老年病研究所,邮编:430065,电话:027-68890008 E-mail:pwang54@aliyun.com
