藏花素通过PI3K/Akt信号途径对阿尔茨海默病细胞模型海马神经元凋亡及P-CREB表达水平的影响  

Effects of Crocin on the Apoptosis of Hippocampal Neurons and P-CREB in Alzheimer′s Disease Cell Model through PI3K/Akt Signaling Pathway

作　　者：杨艳 杨爱明[2] 孙俊 张征 白翔 YANG Yan;YANG Aiming;SUN Jun;ZHANG Zheng;BAI Xiang(Yunnan University of Chinese Medicine,Kunming 650500,Yunnan,China)

机构地区：[1]云南中医药大学,昆明650500 [2]云南省中医医院

出　　处：《中西医结合心脑血管病杂志》2020年第23期3954-3958,共5页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的研究藏花素通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号途径对阿尔茨海默病(AD)细胞模型海马神经元凋亡及磷酸化环磷酸腺苷(cAMP)反应序列结合蛋白(P-CREB)表达水平的影响。方法原代培养大鼠海马神经元,倒置显微镜观察细胞正常生长。海马神经元被分为正常组、藏花素组,其中藏花素组细胞用Aβ25~35孵育制备成AD细胞模型,并加入不同终浓度的藏花素(0μmol、1μmol、3μmol、10μmol),正常组直接加入等体积空白溶剂。采用MTT法检测神经元存活分数,采用TUNEL染色检测海马神经元凋亡情况,蛋白质印迹法(Western Blot)检测PI3K/Akt信号途径相关蛋白(PI3K、Akt、Bcl-2和Bax)以及P-CREB表达水平,荧光定量聚合酶链式反应(RT-PCR)检测PI3K/Akt信号途径相关蛋白mRNA表达水平。结果藏花素组神经元存活分数低于正常组,凋亡细胞数高于正常组,其中经过Aβ25~35处理后,随着藏花素浓度的增加,神经元存活分数增加,凋亡细胞数降低(P<0.05)。藏花素各剂量组PI3K、AKT、Bcl-2、P-CREB蛋白以及mRNA表达水平低于正常组,而Bax蛋白以及mRNA高于正常组,其中经过Aβ25~35处理后,随着藏花素浓度的增加,PI3K、Akt、Bcl-2、P-CREB蛋白以及mRNA表达水平均增加,而Bax蛋白以及mRNA表达水平降低(P<0.05)。结论藏花素可通过调控PI3K/Akt信号途径以及促进P-CREB表达,从而发挥海马神经元保护作用,抑制细胞凋亡。Objective To observe the effects of crocin on apoptosis of hippocampal neurons and phosphorylated cAMP responsive element binding protein(P-CREB)in Alzheimer′s disease(AD)cell model through phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods Primary cultured hippocampal neurons were observed under inverted microscope.Hippocampal neurons were divided into the normal group and the treated group,in which the AD cell model was prepared by incubation with Aβ25~35 in the treatment group,thus added with different final concentrations of crocin(0μmol、1μmol、3μmol、10μmol),and the normal group was directly treated with equal volume blank solvent.The survival fraction of neurons was detected by MTT,the apoptosis of ktocampal neurons was detected by TUNEL staining,the expression of PI3K/Akt signal pathway related proteins(PI3K,Akt,Bcl-2 and Bax)and P-CREB were detected by Western Blotting,and the expression of PI3K/Akt signal pathway related proteins mRNA was detected by RT-PCR.Results The survival fractions of neurons in the treatment group were significantly lower than those in the normal group,and the numbers of apoptotic cells in the treatment group were significantly higher than those in the normal group.After treatment with Aβ25~35,the survival fraction of neurons was increased significantly and the number of apoptotic cells was decreased significantly with the increase of crocin concentration(P<0.05).The protein and mRNA expressions of PI3K,Akt,Bcl-2,and P-CREB in the treatment group were significantly lower than those in the normal group,while the protein and mRNA of Bax were significantly higher than those in the normal group.After treatment with Aβ25~35,the protein and mRNA expressions of PI3K,Akt,Bcl-2,and P-CREB increased significantly with the increase of crocin concentration,while the protein and mRNA of Bax decreased significantly.Conclusion Crocin could protect hippocampal neurons and inhibit cell apoptosis by regulating PI3K/Akt signal pathway and promoting the exp

关 键 词：阿尔茨海默病 环磷酸腺苷反应序列结合蛋白 藏花素 海马神经元 磷脂酰肌醇3-激酶/蛋白激酶B 实验研究 

分 类 号：R749[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]