摘要
目的探讨牛膝提取物含药血清对β淀粉样蛋白_(1-42)(amyloidβ-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。
Objective To explore the effects and mechanism of Achyranthis bidentatae Radix extract-containing serum on neuronal oxidative stress and apoptosis induced by amyloidβ-protein_(1-42)(Aβ_(1-42)).Methods Rat hippocampal neurons were cultured in vitro and divided into control group,model group(Aβ_(1-42) induced 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_(1-42) induced neurons transfected with antisense RNA of zinc finger and BTB domain protein 20 small interfering RNA]and si-NC group(Aβ_(1-42) induced 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 acid-specific 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_(1-42) induced oxidative stress and apoptosis of hippocampal neurons by down-regulating the expression of ZBTB20-AS1.
作者
马宝君
叶涵斌
孟高强
叶子
MA Baojun;YE Hanbin;MENG Gaoqiang;YE Zi(Department of Neurosurgery,The First People's Hospital of Nantong,Nantong,Jiangsu 226001,China)
出处
《湖南中医药大学学报》
CAS
2024年第8期1366-1372,共7页
Journal of Hunan University of Chinese Medicine
基金
江苏省自然科学基金项目(BK20221043)。
关键词
牛膝提取物
β淀粉样蛋白_(1-42)
海马神经元
氧化应激
凋亡
Achyranthis bidentatae Radix extract
amyloid β-protein_(1-42)
hippocampal neurons
oxidative stress
apoptosis
作者简介
通信作者:叶子,男,硕士,主治医师,E-mail:ntsdyrmyymbj@163.com。
