基于垂体腺苷酸环化酶激活多肽在帕金森动物模型中的神经保护作用及机制探索  

Exploring the neuroprotective effects and mechanisms of pituitary adenylate cyclase-activating polypeptides in Parkinson's disease animal models

作　　者：裴丽娟 田洪战 李蕊 田婷[2] 张敏[2] 蔡宏斌[2] Pei Lijuan;Tian Hongzhan;Li Rui;TianTing;Zhang Min;Cai Hongbin(Third People's Hospital of Gansu Province,Lanzhou 730030,Gansu,China;The Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730030,Gansu,China)

机构地区：[1]甘肃省第三人民医院,甘肃兰州730030 [2]兰州大学第二医院(第二临床医学院),甘肃兰州730030

出　　处：《生物医学转化》2024年第1期108-116,共9页Biomedical transformation

基　　金：甘肃省自然科学基金项目(23JRRA0976);兰州大学第二医院萃英学子科研培育计划项目(CYXZ2019-02)。

摘　　要：目的 探讨垂体腺苷酸环化酶激活多肽(PACAP)在帕金森病(PD)动物模型中的神经保护作用及机制。方法SD大鼠设置假手术组(Sham组)、PD组(帕金森造模组)、PD+PACAP_低浓度组(造模+按体重5μg/kg予PACAP鼻腔给药)、PD+PACAP_中浓度组(造模+按体重10μg/kg予PACAP鼻腔给药)、PD+PACAP_高浓度组(造模+按体重20μg/kg予PACAP鼻腔给药) 5组,最终每组纳入10只。四个PD组将脂多糖溶液注射至SD大鼠黑质中建立PD模型,假手术组同法操作但注射生理盐水。通过转棒法检测大鼠行为学表现;ELISA法检测黑质谷氨酸浓度(Glu)和α-突触核蛋白(α-syn)水平,WB法检测黑质α-syn及核因子激活的B细胞的κ-轻链(NF-κB)、核因子κB抑制蛋白α (IκBα)的表达水平;实时荧光定量聚合酶链式反应法(qPCR)检测大鼠黑质炎症因子肿瘤坏死因子α (TNF-α)、白细胞介素-1β (IL-1β)、白细胞介素-6IL-6的mRNA表达水平;WB法和免疫荧光法检测核转录因子胶质纤维酸性蛋白(GFAP)水平来观察星形胶质细胞的活化水平,TUNEL法检测黑质神经元的凋亡情况。结果 (1)行为学结果显示,在转棒实验中,与Sham组比较,PD组潜伏期缩短,掉落次数增加(37.20±5.27次/min);与PD组比较,PACAP组潜伏期延长,掉落次数减少(14.50±2.32次/min),PACAP治疗组的潜伏期和掉落次数随着PACAP浓度的增加而潜伏期可以更长,掉落次数可以更少(P<0.001)。(2)星形胶质细胞激活相关指标显示,PD组GFAP表达水平和黑质神经元的凋亡数量较Sham组显著升高;PACAP治疗组上述指标随着PACAP的浓度增加而显著减低(P<0.01)。(3)黑质神经炎症指标显示,与Sham组比较,PD组NF-κB含量及TNF-α、IL-1β、IL-6的mRNA显著升高,IκBα含量显著降低;PACAP治疗组NF-κB含量及TNF-α、IL-1β、IL-6的mRNA随着PACAP的浓度增加而减低,IκBα含量随着PACAP的浓度增加而升高(P<0.01)。(4) PD疾病特征指标显示,PD组谷氨酸和α-syn含量均较Sham组显Objective To investigate the neuroprotective effects and mechanisms of Pituitary Adenylate Cyclase-Activating Polypeptide(PACAP)in Parkinson's Disease(PD)animal models.Methods SD rats were divided into five groups:sham-operated group(Sham group),PD group(Parkinson's modeling group),PD+PACAP_low concentration group(modeling+5μg/kg of PACAP intranasal administration according to body weight),PD+PACAP_medium concentration group(modeling+10μg/kg of PACAP intranasal administration according to body weight),PD+PACAP_high concentration group(modeling+20μg/kg of PACAP intranasal administration according to body weight).Each group included 10 animals.The four PD models were established by injecting lipopolysaccharide solution into the substantia nigra of SD rats.The Sham group was operated in the same way but injected with saline.The behavioral performance of the rats was assessed using the rotarod test.The concentration of glutamate(Glu)and the levels ofα-synuclein(α-syn)in the substantia nigra were detected using ELISA.The expression levels ofα-syn in the substantia nigra,as well as those ofκ-light chain of nuclear factor-activated B-cells(NF-κB)and inhibitor of nuclear factorκB(IκBα)were detected using WB analysis.Quantitative Polymerase Chain Reaction(qPCR)was used to detect the mRNA expression levels of the rat nigral inflammatory factors Tumor Necrosis Factorα(TNF-α),Interleukin-1β(IL-1β),Interleukin-6(IL-6).WB and immunofluorescence methods were used to detect the levels of nuclear transcription factor Glial Fibrillary Acidic Protein(GFAP)to observe the activation level of astrocytes,and the TUNEL method was used to detect apoptosis in nigrostriatal neurons.Results(1)Behavioral results from the rotarod test indicated that compared to the Sham group,the PD group had a shorter latency and an increased number of falls(37.20±5.27 falls/min).In comparison to the PD group,the PACAP group showed a prolonged latency and a decreased number of falls(14.50±2.32 falls/min).As the concentration of PACAP increas

关 键 词：帕金森病 神经炎症 垂体腺苷酸环化酶激活多肽 星形胶质细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]