【摘要】 目的 探讨牛膝提取物含药血清对β淀粉样蛋白_1-42(amyloid β-protein_1-42,Aβ_1-42)诱导的神经元氧化应激和凋亡的影响及作用机制。方法 体外培养大鼠海马神经元，分为对照组，模型组(Aβ_1-42诱导神经元)，牛膝低、中、高剂量组(15、30、45μg/mL的牛膝提取物含药血清作用于Aβ_1-42诱导的神经元)，si-ZBTB20-AS1组[Aβ_1-42诱导转染锌指和BTB结构域蛋白20的反义RNA小干扰RNA的神经元]和si-NC组(Aβ_1-42诱导转染乱序无意义阴性序列的神经元)。ELISA法检测超氧化物歧化酶（superoxide dismutase,SOD）和丙二醛（malondialdehyde,MDA）含量，CCK-8法检测细胞存活率，流式细胞术检测细胞凋亡率，RT-qPCR法检测ZBTB20-AS1表达水平，Western blot法检测细胞增殖抗原Ki67和胱天蛋白酶-3（cysteine aspartic acid specific protease-3,Caspase-3）表达水平。结果 与对照组比较，模型组SOD含量、细胞存活率、Ki67蛋白表达均降低（P<0.05）,MDA含量、细胞凋亡率、BTB20-AS1表达、Caspase-3蛋白表达均升高（P<0.05）。与模型组比较，牛膝中、高剂量组SOD含量、细胞存活率、Ki67蛋白表达均升高（P<0.05）,MDA含量、细胞凋亡率、BTB20-AS1表达、Caspase-3蛋白表达均降低（P<0.05）。与si-NC组比较，si-ZBTB20-AS1组SOD含量、细胞存活率、Ki67蛋白表达均升高（P<0.05）,MDA含量、细胞凋亡率、BTB20-AS1表达、Caspase-3蛋白表达均降低（P<0.05）。结论 牛膝提取物含药血清可能通过下调ZBTB20-AS1表达抑制Aβ_1-42诱导的海马神经元氧化应激和凋亡。更多还原

【Abstract】 Objective To explore the effects and mechanism of Achyranthis bidentatae Radix extract-containing serum on neuronal oxidative stress and apoptosis induced by amyloid β-protein_1-42(Aβ_1-42).Methods Rat hippocampal neurons were cultured in vitro and divided into control group,model group (Aβ_1-42induced nerves),low-,medium-,and high-dose Achyranthis bidentatae Radix groups (15,30,45μg/mL Achyranthis bidentatae Radix extract-containing serum on Aβ_1-42-induced nerves),si-ZBTB20-AS1 group[Aβ_1-42induced neurons transfected with antisense RNA of zinc finger and BTB domain protein 20 small interfering RNA]and si-NC group (Aβ_1-42induced neurons transfected with disorderly meaningless negative sequence).Content of superoxide dismutase （SOD） and malondialdehyde （MDA） were determined by ELISA;CCK-8 method was used to check the cell survival rate;flow cytometry was used to test the apoptosis rate;RT-qPCR was used to check the expression level of ZBTB20-AS1;Western blot method was used to measure the expression levels of cell proliferation antigen Ki67 and cysteine aspartic acidspecific protease-3 （Caspase-3） protein.Results Compared with control group,SOD content,cell survival rate,and the protein expression of Ki67 in model group were lower （P<0.05）,MDA content,apoptosis rate,the expression of BTB20-AS1,and the protein expression of Caspase-3 were higher （P<0.05）.Compared with the model group,SOD content,cell survival rate,and the protein expression of Ki67 in the middle-and high-dose Achyranthis bidentatae Radix extract groups were higher （P<0.05）,and MDA content,apoptosis rate,the expression of BTB20-AS1,and the protein expression of Caspase-3 were lower （P<0.05）.Compared with the si-NC group,SOD content,cell survival rate,and the protein expression of Ki67 in the si-ZBTB20-AS1 group were higher （P<0.05）,and MDA content,apoptosis rate,the expression of BTB20-AS1,and the protein expression of Caspase-3 were lower （P<0.05）.Conclusion Achyranthis bidentatae Radix extract-containing serum may inhibit Aβ_1-42induced oxidative stress and apoptosis of hippocampal neurons by down-regulating the expression of ZBTB20-AS1.更多还原