摘要
本文通过Aβ25-35诱导体外原代培养的SD乳大鼠海马神经元,建立Aβ毒性损伤细胞模型,结合AnnexinV-FITC/PI荧光双染法流式细胞术、MTT比色法、实时荧光定量PCR及Western blot方法检测川芎嗪(tetrameth-ylpyrazine,TMP)对原代培养的海马神经元细胞活性、早期凋亡率和Bax、Bcl-2基因表达的影响。结果显示川芎嗪高、中剂量可明显增强细胞活性,增加神经元细胞的存活率(P<0.01),可显著抑制海马神经元细胞早期凋亡(P<0.01),抑制凋亡蛋白Bax的表达(P<0.01),增强抗凋亡蛋白bcl-2的表达(P<0.01)。川芎嗪可通过调节Bax/Bcl-2平衡抵抗Aβ25-35诱导的海马神经元凋亡,降低Aβ的神经元毒性,对海马神经元损伤有明显的保护作用。
To evaluate the protective effect of tetramethylpyrazine（TMP）on amyloid β-protein fragment 25-35（Aβ25-35）induced apoptosis in primary culturing hippocampal neurons of SD neonatal rats.Newborn SD rat hippocampal neurons were primarily cultured,then treated with TMP of different doses（10,30,100 mmol/L）and aggregated Aβ25-35（20 μmmol/L）for 24 h.Cell viability were determined by MTT assay.Apoptosis was identified by flow cytometry with annexin V-FITC/propidium iodide（PI）staining and the apoptotic rate was calculated.Real time fluorescent quantitative PCR and Western blot method were employed to detect the transcriptional level（Bax,Bcl-2）and expressions of target proteins（Bax,Bcl-2）in all groups.After exposure to Aβ25-35 for 24 h,the survival rate of hippocampal neurons was remarkably decreased while the cell apoptotic rate was notably increased in Aβ group（Compared with model group P0.01）.Incubation with TMP attenuated the decrease of the survival rate of hippocampal neurons（Compared with model group,10 mmol/L TMP,P0.05;30,100 mmol/L TMP,P0.01）and cell apoptotic rate was decreased（Compared with model group,P0.01）induced by Aβ25-35.Expressions of Bcl-2 were increased,while expressions of Bax was decreased（Compared with model group,10 mmol/L TMP,P0.05;30,100 mmol/L TMP,P0.01）.The ratio of Bax/Bcl-2 decreased in neurons treated with deferent dose of TMP.Hence,it was concluded that TMP can protect hippocampal neurons through inhibiting apoptosis induced by Aβ25-35 and accommodate the ratio of Bax/Bcl-2,which may be the important mechanism of TMP attenuating the neurotoxicity of Aβ25-35.
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2013年第5期684-688,694,共6页
Natural Product Research and Development
基金
安徽省高等学校省级优秀青年人才基金项目(2012SQRL103)
安徽中医学院自然科学研究基金项目(2012zr007)
安徽省高校省级自然科学研究项目(KJ2013Z160)
关键词
川芎嗪
海马神经元
淀粉样蛋白β
细胞凋亡
Bax
Bcl-2
tetramethylpyrazine（TMP）
hippocampal neurons
amyloid β-protein fragment 25-35
apoptosis
Bax
Bcl-2
