牛膝提取物含药血清对Aβ1-42诱导的神经元氧化应激和凋亡的调控机制研究OACSTPCD
Regulatory mechanism of Achyranthis bidentatae Radix extract-containing serum on neuronal oxidative stress and apoptosis induced by Aβ1-42
目的 探讨牛膝提取物含药血清对β淀粉样蛋白 1-42(amyloid β-protein1-42,Aβ1-42)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(Aβ1-42 诱导神经元),牛膝低、中、高剂量组(15、30、45 μg/mL的牛膝提取物含药血清作用于Aβ1-42 诱导的神经元),si-ZBTB20-AS1 组[Aβ1-42 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(Aβ1-42 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 dis-mutase,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,Cas-pase-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Aβ1-42 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 amyloid β-protein1-42(Aβ1-42).Methods Rat hippocampal neurons were cultured in vitro and divided into control group,model group(Aβ1-42 induced nerves),low-,medium-,and high-dose Achyranthis bidentatae Radix groups(15,30,45 μg/mL Achyranthis bidentatae Radix extract-containing serum on Aβ1-42-induced nerves),si-ZBTB20-AS1 group[Aβ1-42 induced neurons transfected with antisense RNA of zinc finger and BTB domain protein 20 small interfering RNA]and si-NC group(Aβ1-42 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 Aβ1-42 induced oxidative stress and apoptosis of hippocampal neurons by down-regulating the expression of ZBTB20-AS1.
马宝君;叶涵斌;孟高强;叶子
南通市第一人民医院神经外科,江苏 南通 226001
中医学
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 extractamyloid β-protein1-42hippocampal neuronsoxidative stressapoptosis
《湖南中医药大学学报》 2024 (008)
1366-1372 / 7
江苏省自然科学基金项目(BK20221043).
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