CAJ | 학술논문

目的探讨乌头酸水合酶(AGO2)在人类精子冷冻复苏前后的表达变化及其致精子功能损伤机制.方法利用计算机精液分析系统(CASA)分析15例冷冻前后人类精子的活力;提取人类冷冻前后精子的总蛋白进行Western blot分析ACO2的含量变化;分析并且进行免疫荧光染色定位ACO2在精子的表达区域;利用化学发光法检测冷冻前后精子三磷酸腺苷(ATP)含量的变化;通过荧光探针JC-1对线粒体鞘染色结果的不同进行线粒体膜电势的检测,同时采用流式细胞仪直接对红色荧光(R1)与绿色荧光(R2)定量分析,比较两组差异.结果与冷冻前组比较,冻后复苏组的精子活力下降了18％,ATP含量下降了32％,线粒体R1的比例下降了27％,差异均有统计学意义(P＜0.05).结论 AC02作为线粒体蛋白在冷冻复苏后发生了蛋白的降解,线粒体的功能受到了损伤,使得ATP的产量下降,导致了精子运动能力缺失.
목적 탐토오두산수합매(AGO2)재인류정자냉동복소전후적표체변화급기치정자공능손상궤제.방법 이용계산궤정액분석계통(CASA)분석15례냉동전후인류정자적활력;제취인류냉동전후정자적총단백진행Western blot분석ACO2적함량변화;분석병차진행면역형광염색정위ACO2재정자적표체구역;이용화학발광법검측냉동전후정자삼린산선감(ATP)함량적변화;통과형광탐침JC-1대선립체초염색결과적불동진행선립체막전세적검측,동시채용류식세포의직접대홍색형광(R1)여록색형광(R2)정량분석,비교량조차이.결과 여냉동전조비교,동후복소조적정자활력하강료18％,ATP함량하강료32％,선립체R1적비례하강료27％,차이균유통계학의의(P＜0.05).결론 AC02작위선립체단백재냉동복소후발생료단백적강해,선립체적공능수도료손상,사득ATP적산량하강,도치료정자운동능력결실.
Objective To investigate the change of aconitase-2 (ACO2) between pre-freeze and post-thaw and possible mechanism of cryo-injury.Methods The parameters of sperm from 15 donaters were analyzed by computer-aided sperm analysis (CASA) and revealed a loss in motility; the Western blotting revealed a reduction of ACO2 content after thawing and ACO2 was located in mitochondrial shealth by immunofluorescence test.The change of adenosine-triphosphate (ATP) contents was determined by Chemiluminescence test.The sperm mitochondrial membrane potential was determined by JC-1 test and indirectly revealed mitochondrial function loss by red/green fluorescence,and quantification of fluorescence was determined by Flow Cytometry.T-test was used to compare these two groups.Results Compared with prefreezing group,the motility decreased 18％,ATP contents decreased 32％ and R1 proportion reduced by 27％ (P ＜ 0.05).Conclusion ACO2,degraded after thawing and resulted in mitochandrial functional iniury and reduction in ATP contents,was responsible for sperm motility loss.