去氧胆酰酪氨酸对人类精子质膜完整性和细胞凋亡的影响  

Effect of deoxycholyltyrosine on human sperm plasma membrane integrity and apoptosis

作　　者：温成丽[1,2] 职瑞娜 王玉柱[2] 黄超[2] 李卫华[2] 丁训城[2] 

机构地区：[1]复旦大学公共卫生学院,上海200032 [2]上海市计划生育科学研究所国家计划生育药具重点实验室,上海200032

出　　处：《中国药理学与毒理学杂志》2014年第3期386-390,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家科技重大专项(2012ZX10001007-009-003);国家科技重大专项(2013ZX10001006-003-002);十二五国家科技支撑计划(2012BAI31B04);国家自然科学基金青年基金项目(81100459);上海市科委项目(13431900800);上海市人口和计划生育委员会(2012JG03)~~

摘　　要：目的探讨去氧胆酰酪氨酸(DCT)对人类精子活力、质膜完整性以及凋亡的影响。方法上游高活力人类精子分别与DCT 0.25,0.5和1.0 g·L-1作用20 min,取精子悬液分析精子活率(SM)和活力(SV);SYBR-14/PI双染后经过荧光显微镜和流式细胞仪观察质膜完整性,AnnexinⅤ/PI双染流式细胞仪分析细胞凋亡。结果 DCT对SM和SV均有显著的抑制作用,且存在量-效关系(rSV=0.91,rSM=0.88,P<0.05)。质膜完整性结果显示,质膜破损精子在DCT 0.25,0.5和1.0 g·L-1时质膜破损精子百分率分别为(54.3±9.8)%,(73.8±3.5)%和(80.6±8.3)%,与空白对照组具有明显差别(P<0.05),且具有明显的量效关系(r=0.91,P<0.05)。凋亡实验结果显示,晚期凋亡精子百分率在0.25,0.5和1.0 g·L-1时,分别是(11.9±1.2)%,(30.1±6.3)%和(75.6±6.3)%,具有明显的浓度依赖性(r=0.98,P<0.05)。DCT≤0.5 g·L-1引起精子早期凋亡率升高,但是高浓度时却出现降低趋势。结论 DCT不仅破坏精子质膜完整性,同时诱发精子凋亡,导致受精能力降低或消失。OBJECTIVE To explore the effect of deoxycholytyosine（ DCT） on human sperm viability,plasma membrane integrity and apoptosis. METHODS Sperms were incubated with DCT 0,0. 25, 0. 5,or 1. 0 g·L^-1for 20 min. The treated sperm suspension was detected by the CASA system to measure the sperm viability（ SV） and sperm motility（ SM）. The treated sperm suspension was also incubated with SYBR-14 /PI and examined by epifluorescence microscopy and flow cytometry（ FCM）. The same treated sperm suspension was cultured by AnnexinⅤ /PI and detected by FCM. RESULTS The sperm viability and motility were significantly inhibited by DCT in a concentration-dependent manner（ rSV= 0. 91,rSM= 0. 88,P 0. 05）. According to the results of sperm plasma membrane tests,the percentage of dead sperm in DCT 0. 25,0. 5,and 1. 0 g·L^-1was（ 54. 3 ± 9. 8） %,（ 73. 8 ± 3. 5） % and（ 80.6 ±8.3） % respectively and had an obvious concentration-effect relationship（ r =0. 91,P 0. 05）. Apoptosis test results showed that the percentage of apoptotic sperm in DCT 0. 25,0. 5,and 1. 0 g·L^-1 was（ 11. 9 ± 1. 2） %,（ 30. 1 ± 6. 3） % and（ 75. 6 ± 6. 3） % respectively,which also had an obvious concentration-effect relationship and was significantly different from the control group（ P 0. 05）. The rate of early-stage apoptosis of sperm increased with DCT at a low concentration（ ≤0. 5 g·L^-1）,but decreased at a high DCT concentration. CONCLUSION DCT can not only destroy the sperm plasma membrane but also induce sperm apoptosis,having an adverse effect on the fertilization ability of sperm.

关 键 词：去氧胆酰酪氨酸 精子获能 细胞凋亡 

分 类 号：R966[医药卫生—药理学]