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目的研究鹿藿正丁醇提取物(n-Butanol extract of Rhynchosia volubilis Lour,BERVL)的体外杀精子作用及机制。同时观察该提取物体外对嗜酸乳酸杆菌的影响,初步评估BERVL作为杀精子剂的可行性和安全性。方法体外杀精子实验以生理盐水为阴性对照,壬苯醇醚(nonoxynolum-9,N-9)为阳性对照,显微镜下观察浓度为250mg/ml,125mg/ml,100mg/ml,90mg/ml,62.5mg/ml的BERVL对人精子活力的影响;体外抑菌实验用试管稀释法和琼脂稀释法;体外杀精子机制探讨:用伊红Y水试验和Hoechst 33342/PI荧光双染,显微镜下检测精子膜功能和精子凋亡。结果 BERVL在90mg/ml浓度组:体外杀精实验,原地活动精子百分率为(4.730±3.231)%;体外抑菌试验,对嗜酸乳酸杆菌抑制率为32.96%~35.70%;伊红Y水溶液染色,精子头部未着色百分率为(7.846±4.706)%。Hoechst 33342/PI荧光双染,凋亡精子百分率为(4.977±2.267)%,死精子百分率为(90.111±4.509)%。结论90mg/ml为BERVL较安全理想的体外杀精浓度。BERVL有显著的体外杀精子作用,可能与破坏精子膜和加速精子凋亡及死亡有关。
Objective To study the in vitro spermicidal effect and mechanism of n-Butanol extract of Rhynchosia volubilis Lour (BERVL). At the same time, observe the effect of the extract on Lactobacillus acidophilus in vitro and preliminarily evaluate the feasibility and safety of BERVL as a spermicide. Methods The in vitro spermicidal test was performed with physiological saline as the negative control. Nonoxynolum-9 (N-9) was used as a positive control. The concentration of the drug was 250mg / ml, 125mg / ml, 100mg / ml and 90mg / 62.5mg / ml of BERVL on human sperm vitality; in vitro antibacterial test tube dilution method and agar dilution method; in vitro spermicidal mechanism: Yishui Y water test and Hoechst 33342 / PI fluorescence double staining, microscopic examination Sperm membrane function and sperm apoptosis. Results In the group of 90mg / ml BERVL, the spermicidal activity in vitro was (4.730 ± 3.231)%. The antibacterial activity in vitro was 32.96% ~ 35.70% Staining, sperm head uncolored percentage (7.846 ± 4.706)%. The percentage of apoptotic sperm was (4.977 ± 2.267)% and the percentage of dead sperm was (90.111 ± 4.509)% with Hoechst 33342 / PI double staining. Conclusion 90mg / ml is safe and ideal for in vitro spermicidal concentration of BERVL. BERVL has a significant in vitro spermicidal effect, which may be related to the destruction of sperm membrane and accelerated sperm apoptosis and death.