铝对大鼠精子质量及精子线粒体的影响
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摘要
目的:研究铝暴露对大鼠精子质量及精子线粒体的超微结构、线粒体膜电位(MMP)水平及膜通透性转换孔(MPTP)功能状态的影响,探讨铝降低精子质量的可能作用机制。方法:取体重为180～200 g雄性Wistar大鼠96只,根据饮水中AlCl_3的半数致死量(LD50),采用AlCl_3·6H_2O水溶液饮水暴露法,设置高剂量组[1/5 LD50,256.72 mg/(kg·d)]、中剂量组[1/10 LD50,128.36 mg/(kg·d)]、低剂量组[1/20 LD50,64.18 mg/(kg·d)]及对照组[0 mg/(kg·d)],每组随机分配24只,连续作用16周。实验结束时对各组大鼠附睾精子进行质量检测,透射电镜下观察精子线粒体结构,并采用流式细胞术测定精子线粒体的MMP水平及MPTP的功能状态。结果:与对照组比较,低、中、高剂量铝暴露组前向运动精子百分率均显著低于对照组[(46.49±5.37)%、(33.50±8.75)%、(16.94±5.00)%vs(66.28±5.68)%,P<0.01];死精子率显著高于对照组[(19.73±5.57)%、(35.80±5.90)%、(55.19±4.97)%vs(12.71±4.84)%,P<0.01];畸形精子百分率显著高于对照组[(19.06±2.44)%、(23.78±3.29)%、(32.06±4.65)%vs(14.56±1.62)%,P<0.01]。随着铝暴露剂量的增加,铝暴露组精子线粒体肿胀越明显;铝暴露组大鼠的精子MMP水平显著低于对照组[(60.88±7.37)%、(51.54±6.12)%、(37.70±7.44)%vs(74.35±4.67)%,P<0.01],MMP水平与铝暴露剂量呈负相关(r_s=-0.819,P<0.01);病理性开放的MPTP水平显著高于对照组[(27.80±5.74)%、(36.58±6.67)%、(64.95±8.07)%vs(15.37±7.13)%,P<0.01],病理性开放MPTP与铝暴露剂量呈正相关(r_s=0.867,P<0.01)。结论:铝暴露可致大鼠精子线粒体肿胀、精子MMP水平降低及MPTP病理性开放,并因此引起精子质量下降。
Objective: To study the impacts of aluminum chloride(AlCl_3) on the sperm quality, sperm mitochondrial membrane potential(MMP) and sperm mitochondrial membrane permeability transition pore(MPTP) function of male rats, and the possible mechanisms of AlCl_3 inducing the declination of sperm quality. Methods: According to the median lethal dose(LD50) of AlCl_3·6 H_2O in drinking water, we randomly assigned 96 male Wistar rats weighing 180-200 g to four groups of equal number and fed them with AlCl_3 aqueous drinking water at 256.72 mg/kg/d(1/5 LD50, high-dose group), 128.36 mg/kg/d(1/10 LD50, medium-dose group), 64.18 mg/kg/d(1/20 LD50, low-dose group) and 0 mg/kg/d(control group), respectively, all for 16 weeks. Then, we examined the quality of the epididymal sperm of the rats, observed the morphology of the sperm mitochondria under the transmission electron microscope, and determined the MMP level of the sperm mitochondria and the function of the MPTP by flow cytometry.Results: The percentage of progressively motile sperm was significantly decreased in the low-, medium-and high-dose AlCl_3 groups as compared with that in the control group([46.49 ± 5.37]%, [33.50 ± 8.75]% and [16.94 ± 5.00]% vs [66.28 ± 5.68]%, P < 0.01), that of dead sperm was remarkably increased([19.73 ± 5.57]%, [35.80 ± 5.90]% and [55.19 ± 4.97]% vs [12.71 ± 4.84]%, P < 0.01), and so was that of morphologically abnormal sperm([19.06 ± 2.44]%, [23.78 ± 3.29]% and [32.06 ± 4.65]% vs [14.56 ± 1.62]%, P < 0.01). Sperm mitochondrial swelling was aggravated in the AlCl_3-exposed rats in a dose-dependent manner. The sperm MMP level was significantly lower in the low-, medium-and high-dose AlCl_3 groups than in the control([60.88 ± 7.37]%, [51.54 ± 6.12]% and [37.70 ± 7.44]% vs [74.35±4.67]%, P < 0.01), with a negative correlation to the dose of AlCl_3(r_s =-0.819, P < 0.01), while the pathologically open MPTP was markedly higher in the former three than in the latter group([27.80 ± 5.74]%, [36.58 ± 6.67]% and [64.95 ± 8.07]% vs [15.37 ± 7.13]%, P < 0.01), with a positive correlation to the dose of AlCl_3(r_s = 0.867, P < 0.01). Conclusion: Exposure to aluminum can cause sperm mitochondrial swelling, decrease the sperm MMP level, induce pathological opening of the MPTP, and consequently reduce sperm quality in male rats.
Objective: To study the impacts of aluminum chloride(AlCl_3) on the sperm quality, sperm mitochondrial membrane potential(MMP) and sperm mitochondrial membrane permeability transition pore(MPTP) function of male rats, and the possible mechanisms of AlCl_3 inducing the declination of sperm quality. Methods: According to the median lethal dose(LD50) of AlCl_3·6 H_2O in drinking water, we randomly assigned 96 male Wistar rats weighing 180-200 g to four groups of equal number and fed them with AlCl_3 aqueous drinking water at 256.72 mg/kg/d(1/5 LD50, high-dose group), 128.36 mg/kg/d(1/10 LD50, medium-dose group), 64.18 mg/kg/d(1/20 LD50, low-dose group) and 0 mg/kg/d(control group), respectively, all for 16 weeks. Then, we examined the quality of the epididymal sperm of the rats, observed the morphology of the sperm mitochondria under the transmission electron microscope, and determined the MMP level of the sperm mitochondria and the function of the MPTP by flow cytometry.Results: The percentage of progressively motile sperm was significantly decreased in the low-, medium-and high-dose AlCl_3 groups as compared with that in the control group([46.49 ± 5.37]%, [33.50 ± 8.75]% and [16.94 ± 5.00]% vs [66.28 ± 5.68]%, P < 0.01), that of dead sperm was remarkably increased([19.73 ± 5.57]%, [35.80 ± 5.90]% and [55.19 ± 4.97]% vs [12.71 ± 4.84]%, P < 0.01), and so was that of morphologically abnormal sperm([19.06 ± 2.44]%, [23.78 ± 3.29]% and [32.06 ± 4.65]% vs [14.56 ± 1.62]%, P < 0.01). Sperm mitochondrial swelling was aggravated in the AlCl_3-exposed rats in a dose-dependent manner. The sperm MMP level was significantly lower in the low-, medium-and high-dose AlCl_3 groups than in the control([60.88 ± 7.37]%, [51.54 ± 6.12]% and [37.70 ± 7.44]% vs [74.35±4.67]%, P < 0.01), with a negative correlation to the dose of AlCl_3(r_s =-0.819, P < 0.01), while the pathologically open MPTP was markedly higher in the former three than in the latter group([27.80 ± 5.74]%, [36.58 ± 6.67]% and [64.95 ± 8.07]% vs [15.37 ± 7.13]%, P < 0.01), with a positive correlation to the dose of AlCl_3(r_s = 0.867, P < 0.01). Conclusion: Exposure to aluminum can cause sperm mitochondrial swelling, decrease the sperm MMP level, induce pathological opening of the MPTP, and consequently reduce sperm quality in male rats.
引文
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[4] 杨颖,凡永,匡延平,等.精子线粒体膜电位与男性精液参数和体质量指数的相关性研究.生殖与避孕,2017,37(4):268-271.
[5] 任大堆,孙竞,陈冬,等.线粒体通透性转换孔对细胞程序性死亡的调控机制及其靶向药物研究进展.国际药学研究杂志,2017,44(5):415-419.
[6] 孔维元,张铁辉.线粒体介导的细胞凋亡与男性不育的研究进展.医学综述,2014,20(22):4058-4060.
[7] Xu F,Liu Y,Zhao H,et al.Aluminum chloride caused liver dysfunction and mitochondrial energy metabolism disorder in rat.J Inorg Biochem,2017,174:55-62.
[8] Perrard MH,Sereni N,Schluth-Bolard C,et al.Complete human and rat ex vivo spermatogenesis from fresh or frozen testicular tissue.Biol Reprod,2016,95(4):89-89.
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[12] Teodoro JS,Palmeira CM,Rolo AP.Mitochondrial membrane potential (ΔΨ) fluctuations associated with the metabolic states of mitochondria.Method Mol Biol,2018,1782:109-119.
[13] Paoli D,Gallo M,Rizzo F,et al.Mitochondrial membrane potential profile and its correlation with increasing sperm motility.Fertil Steril,2011,95(7):2315-2319.
[14] Baines CP,Gutiérrez-Aguilar M.The still uncertain identity of the channel-forming unit(s) of the mitochondrial permeability transition pore.Cell Calcium,2018,73:121-130.
[15] Crompton M.The mitochondrial permeability transition pore and its role in cell death.Biochem J,1999,341 ( Pt 2):233-249.
[16] Solesio ME,Pavlov EV.Mitochondrial calcium uptake in activation of the permeability transition pore and cell death.In Molecular Basis for Mitochondrial Signaling.Springer,2017.107-118.
[17] Mnatsakanyan N,Beutner G,Porter GA,et al.Physiological roles of the mitochondrial permeability transition pore.J Bioenerg Biomembr,2017,49(1):13-25.
[18] 傅龙龙,张林媛,安琪,等.左卡尼汀对冷冻精子运动参数和精子线粒体功能的影响.中华男科学杂志,2018,24(12):1059-1063.
[19] 李然伟,刘睿智,许宗革,等.精子凋亡与精子密度、活力、形态关系探讨.中国实验诊断学,2007,11(6):776-779.
[20] Aziz N,Said T,Paasch U,et al.The relationship between human sperm apoptosis,morphology and the sperm deformity index.Hum Reprod,2007,22(5):1413-1419.
[2] Mouro VGS,Menezes TP,Lima GDA,et al.How bad is aluminum exposure to reproductive parameters in rats?Biol Trace Elem Res,2018,183(2):314-324.
[3] Agnihotri SK,Agrawal AK,Hakim BA,et al.Mitochondrial membrane potential (MMP) regulates sperm motility.In Vitro Cell Dev Biol Anim,2016,52(9):953-960.
[4] 杨颖,凡永,匡延平,等.精子线粒体膜电位与男性精液参数和体质量指数的相关性研究.生殖与避孕,2017,37(4):268-271.
[5] 任大堆,孙竞,陈冬,等.线粒体通透性转换孔对细胞程序性死亡的调控机制及其靶向药物研究进展.国际药学研究杂志,2017,44(5):415-419.
[6] 孔维元,张铁辉.线粒体介导的细胞凋亡与男性不育的研究进展.医学综述,2014,20(22):4058-4060.
[7] Xu F,Liu Y,Zhao H,et al.Aluminum chloride caused liver dysfunction and mitochondrial energy metabolism disorder in rat.J Inorg Biochem,2017,174:55-62.
[8] Perrard MH,Sereni N,Schluth-Bolard C,et al.Complete human and rat ex vivo spermatogenesis from fresh or frozen testicular tissue.Biol Reprod,2016,95(4):89-89.
[9] World Health Organization.Laboratory manual for the examination and processing of human semen.5th ed.Geneva:World Health Organization,2010.16-25.
[10] 陈丽,董君,闫朝君,等.线粒体嵴重构及其调控.生理科学进展,2018,49(1):3-13.
[11] Zorova LD,Popkov VA,Plotnikov EY,et al.Mitochondrial membrane potential.Analyt Biochem 2018,552:50-59.
[12] Teodoro JS,Palmeira CM,Rolo AP.Mitochondrial membrane potential (ΔΨ) fluctuations associated with the metabolic states of mitochondria.Method Mol Biol,2018,1782:109-119.
[13] Paoli D,Gallo M,Rizzo F,et al.Mitochondrial membrane potential profile and its correlation with increasing sperm motility.Fertil Steril,2011,95(7):2315-2319.
[14] Baines CP,Gutiérrez-Aguilar M.The still uncertain identity of the channel-forming unit(s) of the mitochondrial permeability transition pore.Cell Calcium,2018,73:121-130.
[15] Crompton M.The mitochondrial permeability transition pore and its role in cell death.Biochem J,1999,341 ( Pt 2):233-249.
[16] Solesio ME,Pavlov EV.Mitochondrial calcium uptake in activation of the permeability transition pore and cell death.In Molecular Basis for Mitochondrial Signaling.Springer,2017.107-118.
[17] Mnatsakanyan N,Beutner G,Porter GA,et al.Physiological roles of the mitochondrial permeability transition pore.J Bioenerg Biomembr,2017,49(1):13-25.
[18] 傅龙龙,张林媛,安琪,等.左卡尼汀对冷冻精子运动参数和精子线粒体功能的影响.中华男科学杂志,2018,24(12):1059-1063.
[19] 李然伟,刘睿智,许宗革,等.精子凋亡与精子密度、活力、形态关系探讨.中国实验诊断学,2007,11(6):776-779.
[20] Aziz N,Said T,Paasch U,et al.The relationship between human sperm apoptosis,morphology and the sperm deformity index.Hum Reprod,2007,22(5):1413-1419.
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