参芎黄连解毒汤对阿尔茨海默病模型小鼠神经元损伤和Aβ清除的影响  

Effect of Shenxiong Huanglian Jiedu Decoction on Neuronal Damage and AβClearance in Mice Model of Alzheimer's Disease

作　　者：刘晶 陈康[1] 周渝顺 张哲祚 于顾然[1] 李浩 LIU Jing;CHEN Kang;ZHOU Yushun;ZHANG Zhezuo;YU Guran;LI Hao(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China)

机构地区：[1]南京中医药大学附属医院,南京210029 [2]中国中医科学院望京医院,北京100102

出　　处：《中国实验方剂学杂志》2025年第7期43-52,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：科技部国家重点研发计划“中医药现代化”重点专项(2022YFC3501400)。

摘　　要：目的:研究参芎黄连解毒汤对阿尔茨海默病(AD)模型小鼠β淀粉样蛋白(Aβ)清除能力和神经元损伤的影响。方法:选取36只2月龄SPF级C57BL/6J小鼠为实验对象,利用双侧海马区注射Aβ寡聚体的方式构建阿尔茨海默病模型小鼠,设置空白组、假手术组、参芎黄连解毒汤(中药)低、高剂量组(3.27、6.54 g·kg^(-1))和多奈哌齐组(0.65 mg·kg^(-1))。采用水迷宫和旷场实验评价小鼠的学习记忆能力和活动功能,苏木素-伊红(HE)和尼氏染色法观察脑组织病理,活体成像评估荧光Aβ经脑脊液的代谢情况,离子钙结合衔接分子-1(Iba-1)染色观察小胶质细胞活化情况,免疫荧光和酶联免疫吸附测定法检测脑组织和血清中神经营养因子-3(NT-3)和脑源性神经营养因子(BDNF)含量,蛋白免疫印迹法(Western blot)检测海马组织中炎症和通路蛋白的表达。结果:与空白组、假手术组比较,模型组小鼠逃避潜伏期延长,平台停留时间缩短,海马组织出现神经元固缩,尼氏体溶解、小胶质细胞活化等病理表现,荧光Aβ经脑脊液代谢速度减慢,海马组织BDNF、NT-3、白细胞介素-10(IL-10)表达水平显著降低(P<0.01);海马组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、Toll样受体4(TLR4)、髓样分化因子88(MyD88)和磷酸化核转录因子-κB(p-NF-κB p65)表达水平明显升高(P<0.05,P<0.01);与模型组比较,中药低、高剂量组和多奈哌齐组小鼠逃避潜伏期缩短,平台停留时间延长;海马组织神经元核固缩、尼氏体溶解、小胶质细胞活化等情况好转,荧光Aβ经脑脊液代谢增快,海马组织中BDNF、NT-3表达增多(P<0.01),TLR4、MyD88、p-NF-κB p65表达显著降低(P<0.05,P<0.01);中药高剂量组海马组织TNF-α明显降低(P<0.05),IL-10表达明显升高(P<0.05);多奈哌齐组海马组织TNF-α、IL-6、IL-1β的表达明显降低(P<0.05,P<0.01)。结论:参芎黄连解毒能改善AD小鼠神经元损�Objective:To investigate the effects of Shenxiong Huanglian Jiedu decoction on the clearance of amyloidβ-protein(Aβ)and neuronal damage in the mouse model of Alzheimer's disease(AD).Methods:A total of 36 SPF-grade 2-month-old C57BL/6J mice were used in this study,and the modeling was performed by bilateral hippocampal injection of Aβoligomers in C57BL/6J mice.The experiment was conducted with a blank group,a sham operation group,a model group,low-and high-dose(3.27,6.54 g·kg^(-1),respectively)Shenxiong Huanglian Jiedu decoction groups,and a positive control(donepezil hydrochloride,0.65 mg·kg^(-1))group.At the end of the drug intervention,the learning and memory abilities and the activities of mice were evaluated by the Morris water maze and open field tests.Brain histopathology was examined by hematoxylin-eosin and Nissl staining.Additionally,in vivo imaging was employed to measure the metabolism of fluorescent Aβin the cerebrospinal fluid,and staining of ionized calcium-binding adapter molecule-1(Iba-1)was employed to assess microglial activation in the hippocampal tissue.Additionally,neurotrophin-3(NT-3)and brain-derived neurotrophic factor(BDNF)levels in the brain tissue and serum were determined by the immunofluorescence assay and enzyme-linked immunosorbent assay.Western blot was conducted to determine the expression of inflammation and pathway-related proteins in the hippocampal tissue.Results:Compared with the blank group and the sham operation group,the escape latency of the mice in the model group was prolonged,the platform residence time was shortened,the hippocampal tissue showed pathological manifestations such as neuronal pyknosis,Nissl body dissolution,and microglia activation.The metabolic rate of fluorescent Aβthrough cerebrospinal fluid was slowed down,and the expression levels of BDNF,NT-3,and interleukin-10(IL-10)in the hippocampus were significantly decreased(P<0.01).The expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),Toll-like receptor

关 键 词：参芎黄连解毒汤 阿尔茨海默病 神经元损伤 β淀粉样蛋白(Aβ)清除 

分 类 号：R259[医药卫生—中西医结合] R285[医药卫生—中医内科学] R742[医药卫生—中医学]