黄芪多糖调控Nrf2/HO-1通路对阿尔茨海默病大鼠海马神经元损伤的作用机制研究  

Effect of Astragalus Polysaccharide on Hippocampal Neuron Injury in Alzheimer′s Disease Rats

作　　者：范红娟[1] 李书瑞[1] 康凯宁[1] 关连颖[1] FAN Hongjuan;LI Shurui;KANG Kaining;GUAN Lianying(Handan Central Hospital,Handan 056000,Hebei,China)

机构地区：[1]邯郸市中心医院,河北邯郸056000

出　　处：《中西医结合心脑血管病杂志》2023年第23期4340-4346,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的:探讨黄芪多糖(APS)对阿尔茨海默病(AD)大鼠海马神经元损伤的影响及其机制。方法:按随机数字表法将180只健康Wistar大鼠平均分为正常组、模型组、APS低剂量组(APS-L组)、APS中剂量组(APS-M组)、APS高剂量组(APS-H组)和吡拉西坦片组。除正常组外,其他各组采用双侧海马定向注射β-淀粉样蛋白1~42片段(Aβ_(1~42))的方法制备AD大鼠模型,APS-L组、APS-M组、APS-H组分别给予200、400、800 mg/kg APS灌胃,吡拉西坦片组给予500 mg/kg吡拉西坦片灌胃,正常组和模型组给予0.9%氯化钠溶液灌胃,每日1次,连续给予30 d。苏木精-伊红染色观察海马CA1区神经元病理改变,透射电子显微镜观察神经元超微结构,分光光度法检测海马组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,酶联免疫吸附(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6水平,实时荧光定量聚合酶链式反应(RT-PCR)法测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)mRNA表达,蛋白免疫印迹(Western Blot)法检测Nrf2、HO-1、胞浆核因子-κB p65(NF-κB p65)、胞核NF-κB p65蛋白表达。结果:与模型组比较,APS-L组、APS-M组、APS-H组和吡拉西坦片组大鼠海马CA1区神经元病理性形态结构改变、超微结构病变呈不同程度改善。APS-M组、APS-H组和吡拉西坦片组海马CA1区神经元病理分级明显降低(P<0.05),SOD、GSH-Px活性明显升高且MDA含量明显降低(P<0.05),TNF-α、IL-1β、IL-6水平明显降低(P<0.05),Nrf2、HO-1 mRNA和蛋白表达量均明显升高(P<0.05),胞核NF-κB p65表达量和胞核NF-κB p65/胞浆NF-κB p65明显降低(P<0.05)。APS对各检测指标的影响具有一定剂量依赖性,APS-H组对各指标的影响均优于吡拉西坦片组(P<0.05)。结论:APS对AD大鼠海马神经元损伤具有保护作用,该作用具有一定的剂量依赖性,其机制可能与激活Nrf2/HO-1通路、抑制NF-κB核转位进而Objective:To investigate the effects of astragalus polysaccharide(APS)on hippocampal neuron injury in rats with Alzheimer′s disease(AD).Methods:A total of 180 healthy Wistar rats were divided into normal group,model group,low APS(APS-L),medium APS(APS-M),high APS dose(APS-H)group and piracetam(PIR)group by random number table method.Except the normal group,the AD rat model was prepared by bilateral hippocampal injection ofβ-amyloid 1-42 fragments(Aβ1-42).APS-L,APS-M and APS-H groups were given 200,400 and 800 mg/kg APS intragastatically,respectively.The PIR group were given 500 mg/kg PIR intragastatically.The normal group and model group were given 0.9%sodium chloride solution intragastatically,once a day,for 30 days.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of neurons in CA1 region of hippocampus.Transmission electron microscopy was uesd to observe the ultrastructure of neurons.Superoxide dismutase(SOD),glutamate glutathione peroxidase(GSH-Px)activity and malondialdehyde(MDA)content in hippocampus were detected by spectrophotometry.The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)were measured by reverse transcription polymerase chain reaction(RT-PCR).The expressions of Nrf2,HO-1,cytoplasmic nuclear factor-κB p65(NF-κB p65),and nuclear NF-κB p65 were detected by Western Blot.Results:Compared with model group,the pathological morphological and ultrastructural changes of neurons in hippocampal CA1 region of rats in APS-L,APS-M,APS-H group,and PIR group improved with different degrees.The pathological grade of neurons in hippocampal CA1 region significantly decreased in APS-M,APS-H and PIR groups(P<0.05),the activities of SOD and GSH-Px significantly increased and the content of MDA significantly decreased(P<0.05),and the levels of TNF-α,IL-1βand IL-6 significantly decreased(P<0.05),Nrf2 and HO-1 mRNA an

关 键 词：阿尔茨海默病 黄芪多糖 海马神经元 核因子E2相关因子2/血红素加氧酶1通路 氧化应激 

分 类 号：R285.5[医药卫生—中药学]