开心散对双侧海马CA1区注射Aβ_(1-42)致AD大鼠的治疗作用及机制研究  

Therapeutic effect and mechanisms of Kaixin San in AD rats induced by bilateral Aβ_(1-42) injection into the CA1 area of the hippocampus

作　　者：裴海鸾 马贝贝 王婷婷 李瑞吉 刘金辉 于尚玥 娄天宇 左泽平 田时秋 李依林 王晨晓 田颖颖 田骄 赵新月 刘闯 郭玉东 王晶[1] 王志斌 PEI Hailuan;MA Beibei;WANG Tingting;LI Ruiji;LIU Jinhui;YU Shangyue;LOU Tianyu;ZUO Zeping;TIAN Shiqiu;LI Yilin;WANG Chenxiao;TIAN Yingying;TIAN Jiao;ZHAO Xinyue;LIU Chuang;GUO Yudong;WANG Jing;WANG Zhibin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Beijing Tongrentang Research Institute,Beijing 100071;Beijing Institute for Drug Control,Beijing 102200)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]北京同仁堂研究院,北京100071 [3]北京市药品检验研究院,北京102200

出　　处：《中国比较医学杂志》2022年第10期78-90,141,共14页Chinese Journal of Comparative Medicine

基　　金：北京中医药大学科研发展基金(2019-ZXFZJJ-099)。

摘　　要：目的考察开心散对双侧海马CA1区注射Aβ_(1-42)致AD大鼠的治疗效果,初步明晰其作用机制。方法以“β-淀粉样蛋白毒性学说”为依据,使用脑立体定位仪在SD大鼠双侧海马CA1区注射Aβ_(1-42)寡聚肽段,从而复制AD大鼠模型。将痴呆大鼠随机分为模型组、石杉碱甲组、开心散低剂量组、中剂量组与高剂量组,每组10只。另取10只大鼠颅内注射等量的生理盐水为假手术组,正常大鼠10只作为空白对照组。连续灌胃给药30 d。通过Morris水迷宫行为学测试观测大鼠学习记忆成绩的变化;HE染色及Nissl染色观察脑组织病理形态;ELISA法检测脑组织中胆碱能神经递质(ACh、AChE、ChAT)、炎症因子(TNF-α、IL-6、IL-1β)及β-淀粉样蛋白(APP、Aβ_(1-40)、Aβ_(1-42))的含量;IHC法解析皮层、海马组织中tau蛋白及GSK-3β的表达水平;Western blot法察看脑组织中BAX与BCL-2的表达程度。结果与模型组相比,开心散干预组大鼠逃避潜伏期明显缩短,穿越平台次数增多,在目标象限的停留时间及游泳路程所占百分比均有增加(P<0.05或P<0.01);皮层、海马组织神经元数量增多,形态结构趋于正常,且尼氏小体数量增加,胞浆着色变深;脑组织中AChE活性减弱,ChAT及ACh含量明显升高(P<0.05或P<0.01);TNF-α、IL-6、IL-1β、APP、Aβ_(1-40)及Aβ_(1-42)的含量显著下降(P<0.05或P<0.01);皮层、海马组织中tau蛋白及GSK-3β的阳性表达量皆有下调(P<0.05或P<0.01);BAX蛋白的表达水平趋于下降,且BCL-2/BAX的比值有不同程度的升高趋势(P>0.05)。结论开心散对AD损伤大鼠的痴呆症状与脑组织损伤有较好的治疗效果,其作用机制涉及到胆碱能系统的修复、炎症因子释放的抑制、β-淀粉样蛋白的清除及tau蛋白磷酸化水平的调控。Objective To examine the therapeutic effect of Kaixin San in rats with AD caused by bilateral Aβ_(1-42) injection into CA1 area of the hippocampus,and to preliminarily clarify its mechanisms.Methods Using the“β-amyloid protein toxicity theory”,the rat model of AD was established by bilaterally injecting Aβ_(1-42) oligopeptide segments in the CA1 region of the hippocampus of SD rats using a brain stereotaxic instrument.The rats were randomly divided into model,Hupezine A,Kaixin San low,middle and high dose groups,with 10 rats in each group.Another 10 rats were subjected to intracranial injection of an equal amount of physiological saline as the sham operation group and 10 normal rats were used as the blank control.The rats were subjected to continuous intragastric administrations for 30 days.Changes in learning and memory performance were observed by the Morris water maze behavioral test.HE and Nissl staining was used to visualize pathological morphology of the brain.The contents of cholinergic neurotransmitters(ACh,AChE and ChAT),proinflammatory factors(TNF-α,IL-6 and IL-1β),andβ-amyloid protein(APP,Aβ_(1-40) and Aβ_(1-42))in brain tissue were measured by ELISA.Expression of tau protein and GSK-3βin cortical and hippocampal tissues was detected by IHC.BAX and BCL-2 expression in brain tissues was measured by Western blot.Results Compared with the model group,rats in the Kaixin San treatment groups had a significantly shorter escape latency,an increased number of platform crossings,and an enhanced dwell time and percentage of swimming distance to the total distance in the target quadrant(P<0.05 or P<0.01).The numbers of neurons in cortical and hippocampal tissues were increased,the morphological structure tended to be normalized,the number of Nissl bodies increased,and the cytoplasm darkened.AChE activity was attenuated,and ChAT with ACh levels were significantly increased in brain tissue(P<0.05 or P<0.01).TNF-α,IL-6,IL-1β,APP,Aβ_(1-40) and Aβ_(1-42) contents were decreased markedly(P<0.05 or P<0.

关 键 词：开心散 AD大鼠 脑组织 治疗作用 作用机制 

分 类 号：R-33[医药卫生]