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小檗碱对原代大鼠皮质神经元氧化应激的保护作用

The Protective Effect of Jatrorrhizine Against Oxidative Stress in Primary Rat Cortical Neurons.

作者信息

Luo Tao, Shen Xiu-Yin, Li Sheng, Ouyang Ting, Mai Quan-An, Wang Hua-Qiao

机构信息

Department of Anatomy and Neurobiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, 510080, China.

Guangzhou Sport University, Guangzhou 510500, China.

出版信息

CNS Neurol Disord Drug Targets. 2017;16(5):617-623. doi: 10.2174/1871527315666160711101210.

Abstract

OBJECTIVES

This study investigated the neuroprotective effects of Jatrorrhizine in rat cortical neurons.

METHOD

The effects of Jatrorrhizine on hydrogen peroxide (H2O2)-induced cell lesion, levels of lipid peroxidation and antioxidant enzyme activities were investigated in rat cortical neurons. Levels of mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) were measured by fluorescent rhodamine staining and 2',7'-dichlorfluorescein-diacetate staining, respectively. ATP content was measured by a high performance liquid chromatography. The protein levels for Bax, Bcl2 and cleaved caspase-3 were analyzed by western blot protein expression.

RESULTS

There was a significant reduction in cell viability and activities of Superoxide dismutase and glutathione peroxidase for the cortical neurons after exposure to 50μM H2O2 for 12h. The hydrogen peroxide increased the production of malondialdehyde and ROS but decreased MMP and ATP in the neurons. However, pretreatment with different concentrations of Jatrorrhizine (5-20μM) inhibited H2O2-induced neurotoxicity markedly. Jatrorrhizine also attenuated the H2O2-induced Bcl-2/Bax ratio reduction and caspase-3 activation in these neurons.

CONCLUSIONS

Our findings suggest that Jatrorrhizine plays a critical neuroprotective role in H2O2 - induced apoptosis through its anti-oxidative actions. This may allow Jatrorrhizine to be a novel therapeutic with its high bioavailability to treat Alzheimer's disease.

摘要

目的

本研究调查了小檗碱对大鼠皮质神经元的神经保护作用。

方法

研究了小檗碱对过氧化氢(H2O2)诱导的大鼠皮质神经元细胞损伤、脂质过氧化水平和抗氧化酶活性的影响。分别通过罗丹明荧光染色和2',7'-二氯荧光素二乙酸酯染色测量线粒体膜电位(MMP)和细胞内活性氧(ROS)水平。通过高效液相色谱法测量ATP含量。通过蛋白质免疫印迹分析Bax、Bcl2和裂解的caspase-3的蛋白质水平。

结果

暴露于50μM H2O2 12小时后,皮质神经元的细胞活力以及超氧化物歧化酶和谷胱甘肽过氧化物酶的活性显著降低。过氧化氢增加了神经元中丙二醛和ROS的产生,但降低了MMP和ATP。然而,用不同浓度的小檗碱(5-20μM)预处理可显著抑制H2O2诱导的神经毒性。小檗碱还减弱了H2O2诱导的这些神经元中Bcl-2/Bax比值降低和caspase-3激活。

结论

我们的研究结果表明,小檗碱通过其抗氧化作用在H2O2诱导的细胞凋亡中发挥关键的神经保护作用。这可能使小檗碱成为一种具有高生物利用度的治疗阿尔茨海默病的新型疗法。

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