温脾通络开窍方联合头穴透刺对阿尔茨海默病大鼠海马自噬标志物Beclin1、P62和MAP1-LC3的影响  

Effects of Wenpi Tongluo Kaiqiao Prescription combined with scalp acupuncture on autophagy markers Beclin1,P62 and MAP1-LC3 in hippocampus of AD rats

作　　者：陈炜[1,2,3] 匡龙娇 蒋凌飞 朱小敏[4] 洪煌钟 Chen Wei;Kuang Longjiao;Jiang Lingfei;Zhu Xiaomin;Hong Huangzhong(Encephalopathy Department of the First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Key Laboratory of Basic Research of Chinese Medicine,Nanning 530001,China;Graduate School of Jiangxi University of Chinese Medicine,Nanchang 330004,China;Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]广西中医药大学第一附属医院脑病科,南宁530023 [2]广西中医基础研究重点实验室,南宁530001 [3]江西中医药大学研究生院,南昌330004 [4]广西中医药大学研究生院,南宁530001

出　　处：《国际中医中药杂志》2024年第12期1608-1614,共7页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82060844);广西中医药大学重点实验室开放课题(20-065-53-10)。

摘　　要：目的探讨温脾通络开窍方联合头穴透刺法对AD模型大鼠海马自噬标志物自噬关键分子酵母Atg6同系物(Beclin1)、泛素结合蛋白P62(P62)、微血管相关蛋白1轻链3(MAP1-LC3)表达的影响。方法采用脑立体定位注射药物诱导法建立AD大鼠模型,将造模成功大鼠按随机数字表法分为模型组、针药组及西药组,另设假手术组、正常对照组各10只。针药组予头穴透刺联合灌胃温脾通络开窍方16.8 g/kg;西药组灌胃盐酸多奈哌齐片(第1周0.08 mg/kg,第2周0.16 mg/kg,第3周0.24 mg/kg,第4周0.32 mg/kg);假手术组、正常对照组和模型组灌胃等体积生理盐水。造模后2周开始给药,持续给药4周。采用Morris水迷宫测试检测各组大鼠学习记忆能力,采用HE染色及TUNEL染色观察大鼠海马CA1区病理变化,采用Western blot检测海马Beclin1、P62及MAP1-LC3蛋白表达。结果与模型组比较,针药组和西药组大鼠逃避潜伏期明显缩短(P<0.05),穿越平台次数增加(P<0.05)。HE、TUNEL结果提示,模型组大鼠海马CA1区组织结构、细胞形态异常且细胞变性显著,海马神经元细胞质内自噬泡较多,自噬溶酶体结构清晰完整;针药组大鼠海马CA1区组织结构、细胞形态及细胞变性显著改善。针药组和西药组细胞凋亡百分比降低(P<0.05),海马组织Beclin1蛋白表达升高,MAP1-LC3及P62蛋白表达降低(P<0.05)。结论温脾通络开窍方联合头穴透刺法可能通过促进海马Beclin1表达,抑制P62、LC3表达强化自噬水平、提高自噬活性,从而抑制海马CA1区神经元凋亡,促进AD大鼠认知功能和记忆能力恢复。Objective To investigate the effects of Wenpi Tongluo Kaiqiao Prescription combined with scalp acupuncture on the expression levels of hippocampal autophagy markers Beclin1,P62 and MAP1-LC3 in Alzheimer's disease(AD)model rats.Methods The AD rat model was established by stereotactic drug injection,and randomly divided into model group,acupuncture-medicine group,and Western medicine group.Sham-operation group and control group were set,with 10 rats in each group.The acupuncture-medicine group was treated with scalp acupuncture plus Wenpi Tongluo Kaiqiao Prescription(16.8 g/kg);the Western medicine group was treated with donepezil hydrochloride tablets(0.08 mg/kg in week 1,0.16 mg/kg in week 2,0.24 mg/kg in week 3,0.32 mg/kg in week 4)for gavage.The sham-operation group,the control group and the model group were treated with equal volume of normal saline for gavage.The drug was administered 2 weeks after modeling.Morris water maze was used to test the learning and memory abilities of rats in each group after continuous administration for 4 weeks,and the hippocampus of the animal was taken after Morris water maze.HE staining and TUNEL staining were used to observe the pathological changes of the hippocampal CA1 region in each group.Western blot was used to detect the expression levels of Beclin1,P62 and MAP1-LC3 protein genes.Results Compared with the model group,the escape latency of rats in the acupuncture-medicine group and Western medicine groups was significantly shortened(P<0.05),and the number of times they crossed the platform increased(P<0.05).The HE and TUNEL results indicated that the model group rats had abnormal tissue structure and cell morphology in the CA1 area of the hippocampus,with significant cell degeneration.There were more autophagic vesicles in the cytoplasm of hippocampal neurons,and the structure of autophagic lysosomes was clear and complete;the tissue structure,cell morphology,and cellular degeneration of the CA1 area in the hippocampus of rats treated with acupuncture and medicine were s

关 键 词：阿尔茨海默病 温脾通络开窍方 头穴透刺 自噬 认知障碍 BECLIN1 大鼠 

分 类 号：R285.5[医药卫生—中药学]