苍术苷体外对大鼠精子线粒体膜电位及凋亡的影响

Effects of Atractyloside (ATR) on the Mitochondrial Membrane Potential and Apoptosis in Rat Sperm in Vitro

目的:观察中药苍耳子的毒性成分苍术苷(Atractylodesin,ATR)体外对大鼠精子线粒体膜电位、超微结构及凋亡的影响,分析苍耳子可能存在的生殖毒性。方法:取雄性SD大鼠,麻醉、分离附睾精子,分别与不同浓度苍术苷(1.51、3.02、6.04、12.08 mmol/L)共同孵育。观察精子线粒体通透性开放情况,经流式细胞仪检测精子线粒体膜电位(MMP)和凋亡,并在电子显微镜下观察精子线粒体超微结构的改变。结果:与空白对照组比较,苍术苷(3.02 mmol/L^12.08 mmol/L)组大鼠精子线粒体膜电位明显下降,精子早期凋亡率和总凋亡率显著增加。电镜观察可见,与苍术苷共同孵育后,大鼠精子线粒体出现肿胀、融合,基质缺失,外膜破裂等现象,并呈现明显剂量依赖性。结论:苍术苷(3.02 mmol/L^12.08 mmol/L)体外能明显促进大鼠精子凋亡和降低线粒体膜电位,改变精子线粒体超微结构,促进线粒体通透性转换孔(mPTP)开放。说明苍耳子可能存在一定的生殖毒性,苍术苷为其生殖毒性的物质基础。

Objective:To investigate the effects of Atractylodesin(ATR),a toxic component of Xanthium sibiricum,on mitochondrial membrane potential,ultra structure and apoptosis of rat sperm in vitro,and to analyze the possible reproductive toxicity of Xanthium sibiricum.Methods:Male SD rats were anesthetized,epididymal sperm were isolated and incubated with different concentrations of atractyloside(12.08,6.04,3.02,1.51 mmol/L).The sperm mitochondrial membrane potential(MMP)and apoptosis were detected by flow cytometry,and the ultra structural changes of sperm mitochondria were observed under an electron microscope.Results:Compared with the control group,sperm mitochondrial membrane potential in Atractyloside group(3.02~12.08 mmol/L)was decreased,the early apoptotic rate and total apoptotic rate in Atractyloside group were increased.Electron microscopic observation showed that after co incubation with atractyloside,rat sperm mitochondria appeared swelling,fusion,matrix loss,adventitia rupture and other phenomena,in significant dose dependent manner.Conclusion:Atractylodesin(3.02~12.08 mmol/L)can promote sperm apoptosis,decrease mitochondrial membrane potential in vitro,change mitochondrial ultra structure of sperm and promote mitochondrial permeability transition pore(mPTP)opening.It suggests that Xanthium sibiricum may present the reproductive toxicity,and Atractylodesin is the material basis of reproductive toxicity.