桔梗皂苷D的杀精效果及促凋亡研究

摘要

目的:研究中药桔梗的单体化学成分桔梗皂苷D(PD)对人精子的杀伤效果,初步探索其杀精机制.方法:将同一健康男性的精液均分为4份,分别用生理盐水、0.15mmol/L PD、0.20mmol/L PD及0.31 mmol/L壬苯醇醚-9(N-9),阳性对照处理样本2min.用计算机辅助的精液分析系统(CASA)检测PD对人精子的杀伤效果,并通过AnnexinV-FITC检测精子凋亡.结果:生理盐水对照组、0.15mmol/L PD组、0.20mmol/L PD组、N-9对照组的精子活率分别为67.97±21.76％,36.68±28.23％,24.18±7.72％,42.74±25.62％.各处理组与生理盐水对照组间差异有统计学意义(P＜0.001),N-9与两个PD剂量组之间差异无统计学意义(P＞0.05).细胞凋亡检测结果显示,PD主要引起精子的晚期凋亡,0.15mmol/L PD、0.20mmo]/L PD、0.25mmol/L PD组的晚期凋亡指数分别为181.7 ±46.0、246.0±52.8、272.8±57.3,PD各剂量组的晚期凋亡率与对照组之间差异有统计学意义(P ＜0.001).结论:PD对人精子有显著的瞬间杀灭效应,它主要引起精子的晚期凋亡,可能与PD对精子膜的损伤有关.PD将来可能作为一种有潜力的临床杀精剂.%Objective:To evaluate the spermicidal and apoptosis-inducing activities of Platycodon D (PD),a major component of triterpene saponins in Platycodon grandiflora (P.grandiflora).Methods:Sperm samples were collected from healthy men aged 23-41 years,and were prepared by gradient centrifugation.Each sperm sample was divided into four groups based on incubation solutions:control (0.9％ sodium chloride),0.15 mmol/L PD,0.20 mmol/L PD,and N-9 control (0.20mg/ml Nonoxynol-9,0.31mmol/L).The sperm motility and viability were assessed by computer-assisted sperm a nalysis system (CASA).The sperm apoptosis was determined by Annexin V-FITC flowcytometry.Results:The sperm viabilities were 67.97 ± 21.76％ for the control group,36.68 ±28.23％ for 0.15 mmol/L PD group,24.18 ± 7.72％ for 0.20mmol/L PD group,and 42.74 ± 25.62％ for N-9 control group.The sperm viability of the control group was significantly higher than those of PD groups (P ＜ 0.001).There were no significant differences in the sperm viability among PD groups and N-9 control group (P ＞ 0.05).A dosage dependence of PD on apoptosis was observed.Apoptosis index of 0.15 mmol/L PD group,0.20 mmo]/L PD group and 0.25 mmol/L PD group were 181.7 ± 46.0,246.0-± 52.8 and 272.8 ± 57.3,respectively.There were significant differences in the apoptosis index among PD groups and the control group (P ＜0.001).Conclusion:These results suggest that PD has significant spermicidal activity,and could be developed as a potential vaginal contraceptive for future use in humans.