吴珍张亚贝云成沈萍萍△.微囊藻毒素-LR 对小鼠肝细胞线粒体功能的影响*[J].,2014,14(2):226-229 |
微囊藻毒素-LR 对小鼠肝细胞线粒体功能的影响* |
Exploring the Effects of MC-LR on Mitochondrial Functionin Mouse Liver* |
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DOI: |
中文关键词: 微囊藻毒素-LR 线粒体膜电位 线粒体自噬 |
英文关键词: Microcystin-LR Mitochondiral membrane potential Mitophagy |
基金项目:江苏省自然科学基金项目(BK2010049);国家自然科学基金项目(91013015) |
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中文摘要: |
摘要目的:探究微囊藻毒素-LR 对小鼠肝细胞线粒体功能的影响。方法:采用BALB/c 小鼠作为模型动物,随机分为3 组:A 组,
空白对照组,正常饮用水;B 组,添加5 g/L微囊藻毒素-LR 的饮用水;C 组,添加30 g/L 微囊藻毒素-LR 的饮用水。分组喂养3
个月,分离小鼠肝脏、提取线粒体,采用线粒体荧光探针JC-1 测定线粒体膜电位(MMP),qRT-PCR检测自噬相关基因Beclin1 和
Lc3琢的转录水平,Western Blot检测细胞色素C的释放,电镜观察线粒体的形态和内部结构。结果:微囊藻毒素-LR 处理组的小
鼠肝细胞线粒体膜电位明显下降,自噬相关基因Lc3琢的转录水平上升,细胞色素C由线粒体释放到胞浆,电镜观察线粒体形态
异常、内部结构被破坏。结论:微囊藻毒素-LR 对小鼠肝细胞线粒体有较强的毒性作用,并引发线粒体自噬。 |
英文摘要: |
ABSTRACT Objective:To investigate the effects of MC-LR on mitochondria isolated from mouse liver. Methods:The mice were
randomly divided into 3 groups: Group A were fed with tap water, Group B were fed with the water containing 5 μg/L MC-LR, while
Group C were fed with water containing 30 μg /L MC-LR for 3 months respectively. Mitochondria were isolated from mouse liver, and
mitophagy was detected through mitochondrial membrane potential by using JC-1, which was widely used in apoptosis studies to monitor
mitochondrial health. After extracting RNA from the mouse liver, transcriptional levels of autophagy-related genes Beclin1 and Lc3α
were measured by qRT-PCR. The release of cytochrome C from mitochondria to plasma was detected by Western Blot. The morphology
of mitochondria was observed by electron microscope. Results:Compared with the mitochondrial uncoupler carbonyl cyanide
m-chlorophenylhydrazone (CCCP), mitochondrial membrane potential declined obviously. The transcriptional level of Lc3αincreased
correspondingly, while that for Beclin1 was not significant. Cytochrome C was released from mitochondria to the plasma. The
abnormality of mitochondria could be detected clearly by electron microscope, and some mitochondria were tending to fall apart.
Conclusion:Mitophagy could be induced in mouse liver after the treatment of MC-LR, which indicated that MC-LR had severe
mitochondrial toxicity in mouse liver. |
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