Bejarano Ignacio, Lozano Graciela M, Ortiz Agueda, García Juan F, Paredes Sergio D, Rodríguez Ana B, Pariente José A
Department of Physiology, Faculty of Science, University of Extremadura, Badajoz, Spain.
Fertil Steril. 2008 Oct;90(4 Suppl):1340-7. doi: 10.1016/j.fertnstert.2007.08.069. Epub 2007 Dec 26.
To determine the role of calcium signaling on apoptosis evoked by the reactive oxygen species H2O2 and by the physiological agonist P in human ejaculated spermatozoa.
Laboratory study.
Center for assisted human reproduction in a hospital in Spain.
PATIENT(S): Forty-five healthy volunteers.
INTERVENTION(S): Spermatozoa were treated with increasing concentrations of hydrogen peroxide (H2O2; 10 microM, 100 microM, and 1 mM) or with 20 microM of P for 5-120 minutes.
MAIN OUTCOME MEASURE(S): Activation of caspase-3 and -9 as well as phosphatidylserine externalization were examined in human ejaculated spermatozoa by fluorescence methods.
RESULT(S): Hydrogen peroxide and P induced activation of caspase-3 and -9. In addition, the effect of H2O2 and P was time dependent. Dimethyl-1,2-bis (aminophenoxy) ethane-N,N,N',N'-tetraacetic acid loading was able to inhibit H2O2- and P-induced caspase-3 activation and phosphatidylserine externalization. Pretreatment of spermatozoa with Ru360, to block the calcium uptake into mitochondria, also was able to decrease the activation of caspase-3 and phosphatidylserine exposure that was stimulated by either H2O2 or P.
CONCLUSION(S): These findings suggest that H2O2- and P-induced mitochondrial apoptosis is dependent on calcium signaling.
确定钙信号在活性氧物质过氧化氢（H2O2）和生理激动剂P诱导的人类射出精子凋亡中的作用。
实验室研究。
西班牙一家医院的人类辅助生殖中心。
45名健康志愿者。
用浓度递增的过氧化氢（H2O2；10微摩尔、100微摩尔和1毫摩尔）或20微摩尔的P处理精子5至120分钟。
采用荧光法检测人类射出精子中半胱天冬酶-3和-9的激活以及磷脂酰丝氨酸外翻情况。
过氧化氢和P诱导了半胱天冬酶-3和-9的激活。此外，H2O2和P的作用具有时间依赖性。二甲基-1,2-双（氨基苯氧基）乙烷-N,N,N',N'-四乙酸负载能够抑制H2O2和P诱导的半胱天冬酶-3激活以及磷脂酰丝氨酸外翻。用Ru360预处理精子以阻断钙进入线粒体，也能够降低由H2O2或P刺激引起的半胱天冬酶-3激活和磷脂酰丝氨酸暴露。
这些发现表明，H2O2和P诱导的线粒体凋亡依赖于钙信号。
