摘要
目的:探讨补肾化痰复方对APP/PS1双转基因阿尔兹海默病(AD)模型小鼠海马区核仁应激及其下游MDM2-P53通路的影响。方法:AD小鼠分为模型组、西药组、中药低剂量组和高剂量组。正常组由同品系非转基因小鼠组成。西药组用盐酸多奈哌齐干预,剂量为0.65 mg·kg^(-1)·d^(-1)。中药高剂量组用补肾化痰复方干预,剂量为6.5 g·kg^(-1)·d^(-1),中药低剂量组的剂量减半。正常组和模型组给与等体积的ddHO。药物干预90 d后,取各组小鼠海马组织,实时荧光定量PCR检测rRNA转录水平;重亚硫酸盐测序法检测rDNA启动子区域甲基化水平;免疫荧光染色检测核仁形态;Western Blot检测核糖体蛋白(RPL5和RPL11)、MDM2,P53和pP53的表达量。结果:与正常组比较,AD模型组小鼠海马区表现出显著的核仁应激反应,包括rRNA转录水平降低、rDNA启动子区域甲基化水平升高、核仁体积减小和阳性细胞率降低(P<0.01),核糖体蛋白(RPL5和RPL11)、MDM2、P53和pP53蛋白表达量升高(P<0.01,P<0.05)。与盐酸多奈哌齐比较,补肾化痰复方能更加有效地缓解AD小鼠的核仁应激压力和逆转上述各项变化。结论:补肾化痰复方能缓解AD诱发的核仁应激压力,并能借助于MDM2-P53通路调控细胞命运,从而缓解AD进程。
Objective:To explore the effects of Bushen Huatan Compound on the nucleolar stress and its downstream MDM2-P53 pathway in the hippocampus of APP/PS1 double transgenic Alzheimer’s disease (AD) model mice.Methods:AD mice were divided into model group,Western medicine group,low-and high-dose group of traditional Chinese medicine.The normal group consisted of non-transgenic mice of the same strain.The Western medicine group was intervened with Donepezil at a dose of0.65 mg·kg^(-1)·d^(-1).The high-dose group of traditional Chinese medicine was intervened by Bushen Huatan Compound at a dose of 6.5 g·kg^(-1)·d^(-1),and the dose of the low-dose group of traditional Chinese medicine was halved.Normal group and model group were given an equal volume of ddHO.After 90 days of drug intervention,the hippocampal tissues of the mice in each group were taken out to complete the following analysis:real-time fluorescent quantitative PCR to detect rRNA transcription levels;bisulfite sequencing to detect the methylation levels of rDNA promoter region;immunofluorescence staining to detect nucleolar morphology and Western Blot to detect the expression levels of ribosomal proteins (RPL5 and RPL11),MDM2,P53 and pP53.Results:Compared with the normal group,the hippocampus of the mice in AD model group showed a significant nucleolar stress response,including decreased rRNA transcription,increased methylation in the rDNA promoter region,decreased nucleolar volume and rates of positive cells (P<0.01),and increased expression levels of ribosomal proteins (RPL5 and RPL11),MDM2,P53 and pP53 (P<0.01,P<0.05).Compared with Donepezil,the Bushen Huatan Compound can alleviate the nucleolar stress and reverse the above-mentioned changes more effectively in the AD mice.Conclusion:The Bushen Huatan Compound can relieve the nucleolar stress induced by AD,and can regulate cell fate with the help of MDM2-P53 pathway,thereby alleviating the process of AD.
作者
黄敏
谭爱华
谢光璟
赵敏
王平
HUANG Min;TAN Ai-hua;XIE Guang jing;ZHAO Min;WANG Ping(Institute of Gerontology,Hubei University of Chinese Medicine,Wuhan 430065,China;School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2022年第10期5638-5642,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
湖北省自然科学基金项目(No.2017CFB383)。
