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马宝君, 叶涵斌, 孟高强, 叶子.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[J].湖南中医药大学学报英文版,2024,44(8):1366-1372.[Click to copy
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| 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 |
| 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) |
| 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-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 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Aβ1-42诱导的海马神经元氧化应激和凋亡。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2024.08.003 |
| Received:June 12, 2024 |
| 基金项目:江苏省自然科学基金项目（BK20221043）。 |
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| Regulatory mechanism of Achyranthis bidentatae Radix extract-containing serum on neuronal oxidative stress and apoptosis induced by Aβ1-42 |
| MA Baojun, YE Hanbin, MENG Gaoqiang, YE Zi |
| (Department of Neurosurgery, The First People's Hospital of Nantong, Nantong, Jiangsu 226001, China) |
| 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-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. |
| Key words: Achyranthis bidentatae Radix extract amyloid β-protein1-42 hippocampal neurons oxidative stress apoptosis |
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