摘要
目的:观察蒺藜皂苷(TTLS)对缺氧/复氧诱导的大鼠皮层神经元凋亡及细胞内游离钙离子浓度变化的影响。方法:原代培养大鼠皮层神经元,建立缺氧/复氧损伤的皮层神经元凋亡模型。比色法测定细胞乳酸脱氢酶(LDH)释放漏出率;AnnexinV-FITC/PI双染,流式细胞仪定量分析细胞凋亡百分率;Fluo-3荧光染色,激光扫描共聚焦显微镜观察细胞内钙平均荧光值变化。结果:缺氧3h复氧12h可诱导出皮层神经元凋亡,引起细胞内钙离子浓度增高,高于对照组(P<0.01)。蒺藜皂苷(0.025g/L)能明显降低缺氧/复氧诱导的神经元凋亡,并可降低细胞内钙浓度,与模型组比较有显著差异(P<0.01)。结论:蒺藜皂苷可降低由缺氧/复氧诱导的皮层神经元的凋亡,减轻细胞损伤,这种作用机制可能与其抑制神经细胞内钙超载有关。
AIM: To study the effect of Tribulus terrestris L. saponin (TTLS) on apoptosis and changes in cytosolic calcium concentration induced by hypoxia/re - oxygenation in rat cortical neurons. METHODS: Rat cortical neurons in primary culture were used, and a apoptosis model was induced by hypoxia/reoxygenation. LDH releasing rate was detected by spectrophotometry. The apoptosis rate of cortical neurons was analyzed quantitatively by flow cytometry with Annexin V - FITC and PI staining. Intracellular free Ca^2+ ( [ Ca^2+ ] i ) was observed with a confocal laser - scanning microscope and determined by mean fluorescent value with Fluo -3 fluometry. RESULTS: Compared to control group, three hours of hypoxia and twelve hours of reoxygenation group induced cortical neuronal apoptosis and significantly increased the intracellular free Ca^2+ concentration(P 〈0.01 ). Compared with model group, TTLS decreased the percentage of neuronal apoptosis and reduced neuronal [ Ca^2+],(P 〈0.01 ). CONCLUSION: TTLS could obviously reduce hypoxia/reoxygenation - induced apoptosis and alleviate the damage degree of rat cortical neurons. The mechanism might be related to inhibiting the calcium overload induced by hypoxia/reoxygenation in rat cortical neurons.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第5期945-949,共5页
Chinese Journal of Pathophysiology
基金
教育部新世纪优秀人才支持计划(No.NCET-05-0139)
吉林敖东洮南药业股份有限公司开发基金资助项目
关键词
蒺藜
神经元
细胞凋亡
细胞内钙
Tribulus terrestris L.
Neurons
Apoptosis
Calcium