摘要
目的:观察蒺藜皂苷(Tribulus terrestris L.saponion,TTLS)对大鼠皮层神经元经缺氧-复氧后细胞凋亡的影响,并探讨其可能的作用机制。方法:离体培养大鼠皮层神经元,先用95%N2与5%CO2混合气体造成细胞缺氧3h,再给予95%O2与5%CO2混和气复氧12h,建立缺氧-复氧损伤的皮层神经元凋亡模型。中药组于缺氧-复氧时均加入TTLS进行干预。Annexin V-FITC/碘化丙啶(propidium iodide,PI)双染,流式细胞仪定量分析细胞凋亡百分率;JC-1荧光染色,激光扫描共聚焦显微镜观察细胞线粒体膜电位的变化,荧光酶标仪测定神经元胞浆内caspase-3/7的活性,并采用免疫细胞化学法观察细胞Bax蛋白表达情况。结果:细胞缺氧3h复氧12h后,凋亡率明显增加,线粒体膜电位显著降低,胞浆内caspase-3/7活性增多,Bax蛋白大量表达(P<0.01)。TTLS能抑制缺氧-复氧损伤的皮层神经元线粒体膜电位的降低,减少caspase-3/7活性,并可降低Bax蛋白的表达,从而抑制细胞凋亡的发生。结论:缺氧-复氧可以诱导皮层神经元的凋亡。TTLS可降低缺氧-复氧诱导细胞凋亡的发生,其机制与维持线粒体膜电位的稳定,抑制caspase酶活性,降低Bax蛋白表达有关。
Objective： To observe the effects of Tribulus terrestris L. saponion （TTLS） on apoptosis in cortical neurons induced by hypoxia-reoxygenation in rats. Methods： Primary culture of rat cortical neurons was performed in vitro. A model of apoptosis of cortical neurons was established by hypoxia and reoxygenation. Hypoxia for 3 h in neural cells was induced with mixture of 95% Nzand 5% CO2, and then reoxygenation in neural cells was induced with mixture of 95% O2 and 5% CO2 for 12 h. Different concentrations of TTLS were administered to traditional Chinese herbal medicine-treated group separately during hypoxia and reoxygenation. The apoptosis rate was analyzed quantitatively by flow cytometry with Annexin V-FITC and propidium iodide staining. Mitochondria membrane potential was observed by a confocal laser-scanning microscope with JC-1 fluorescence. Caspase-3/7activity in cytoplasm was measured by fluorescent plate reader. Bax protein expression was observed by immunohistochemical technique. Results： The percentage of apoptosis was significantly increased, mitochondria membrane potential was obviously decreased, fluorescence of caspase-3/7 activity was increased, and Bax protein was abundantly expressed followed by 3 h of hypoxia and 12 h of reoxygenation （P〈0.01）. TTLS could inhibit the depression of membrane potential induced by hypoxia and reoxygenation, decrease the activity of caspase-3/7, reduce the expression of Bax protein, and inhibit the apoptosis of the cortical neurons. Conclusion： Hypoxia and reoxygenation can induce apoptosis of rat cortical neurons. TTLS can decrease the apoptosis induced by hypoxia and reoxygenation. The mechanism might be related to stabilization of mitochondria membrane potential, inhibition of caspase activity and reduction of Bax protein expression.
出处
《中西医结合学报》
CAS
2008年第1期45-50,共6页
Journal of Chinese Integrative Medicine
基金
教育部新世纪优秀人才支持计划(NoNCET-05-0139)
吉林敖东洮南药业股份有限公司开发基金资助
关键词
蒺藜
神经元
细胞凋亡
线粒体
缺氧
Tribulus terrestris
neurons
apoptosis
mitochondria
anoxia
