In vitro tests for essential sperm functions using the phyto-oestrogen genistein as a test substance

Sperm motility, binding of spermatozoa to the zona pellucida and induction of the acrosome reaction are prerequisites for successful oocyte fertilization. Examination of the physiological and nonphysiological effects of particular compounds on sperm functions requires high-quality in vitro test systems. In this short methodological overview, a reliable combined in vitro test system with bovine gametes is described. The purpose of the study was to evaluate whether aliquots of pooled post-thaw spermatozoa are suitable for examination of environmental substances that affect essential sperm functions. The combined test system includes a number of known methods for the assessment of sperm vitality and motion parameters, acrosomal status, inducibility of acrosome reaction and sperm zona pellucida binding. First observations indicate that genistein inhibits the induction of acrosomal exocytosis and binding of spermatozoa to the zona pellucida. Motility parameters and the viability of bovine spermatozoa were not affected by this substance. It is concluded that genistein, a phyto-oestrogen which is abundant in several plants, can be used as a test substance for the evaluation of effects upon essential bovine sperm functions in vitro.     

葛根药材HPLC指纹图谱与6种成分含量测定研究

目的 建立葛根Pueraria lobate药材HPLC指纹图谱，并测定不同产地葛根中6种成分(3’-羟基葛根素、葛根素、3’-甲氧基葛根素、葛根素芹菜糖苷、大豆苷、大豆苷元)的含量，为葛根优质药材产地与质量控制提供参考。方法 采用HPLC法，葛根药材30%乙醇提取液的分析采用Hypersil BDS C₁₈色谱柱(250 mm×4.6 mm,5μm);0.1%甲酸水-乙腈为流动相，梯度洗脱；体积流量1.0 mL/min；检测波长250 nm；柱温25℃。建立32批葛根药材指纹图谱，并测定不同产地葛根药材6种成分的含量。结果 建立了葛根药材HPLC指纹图谱，共提取出10个共有峰，指认6个共有峰；建立了含量测定方法，并对不同产地来源的32批葛根药材中6种成分含量测定，不同产地葛根质量对比综合评分结果显示河南最高，其次为陕西。结论 建立了葛根HPLC指纹图谱方法，通过对比，各样品与对照图谱相似度均大于0.9，为葛根道地药材质量标准提供整体质量控制依据。结合产地调研信息，可将葛根优质原料药材产地优选为陕西、河南秦岭山脉。