桔梗皂苷D的杀精效果及促凋亡研究

Effects of Platycodin D on spermcidal and apoptosis-inducing activities

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/LPD、0.20mmol/LPD及0.31mmol/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/LPD组、0.20mmol/LPD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/LPD、0.20mmol/LPD、0.25mmol/LPD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 compo- nent 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, 0.31retool/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 via- bilities 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 0.20 mmol/L PD group, and 42.74 ± 25.62% for N - 9 control group. The sperm viability of the control group was significandy 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 mmol/L PD group and 0.25 mmol/L PD group were 181.7±46.0,246.0 ±52.8 and 272.8 ±57.3, respective- ly. 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.