目的:观察和比较自白芸豆中提取的植物凝集素(PHA)对人、小鼠、牛、山羊和兔精子的凝集作用和整体给药的避孕和抗早孕作用。方法:体外实验中,在精液中加入不同浓度的PHA,对使不同物种精子发生凝集作用影响因素进行观察,高浓度PHA制动的精子用相应获能液洗涤后复苏;在体内实验中,对交配见阴栓后的雌鼠于不同时间一次性经阴道给予PHA;另外不同时间间隔对正常性周期小鼠多次给予PHA观察其避孕效果。结果:体外实验结果表明,人、小鼠和牛的精子在低浓度PHA(15.6~125μg/ml)作用下30min就完全发生凝集,但不会致死,不同物种的精子凝集方式有所不同。凝集后的精子可以在室温下(22±7℃)存活7~12h。而≥250μg/ml的高浓度PHA会使这5种精子完全制动并发生不可逆致死。并且在低pH值时,精子存活时间明显缩短。体内实验结果表明经阴道给予PHA具有抗早孕的效果;每天经阴道给予PHA具有100%的避孕效果。结论:在体外实验中,云南大白芸豆植物凝集素具有广泛的精子凝集作用;在体实验证明PHA可有效用于避孕和抗早孕,因此PHA有可能开发成为具有外用避孕作用的药物。
Objective:To observe the impact of phytohemagglutinin(PHA)extracted from Phaseolus vulgaris(white kidney bean)on spermatozoa of different species in vitro and its contraceptive and anti-early pregnancy effectiveness in vivo.Methods:The spermatozoa from human,mice,cattle,goat and rabbit were prepared respectively in their capacitating mediums before different concentrations of PHA were added.The effective concentration of PHA and the duration of sperm aggregation were observed and recorded.The female mice with vaginal plug or normal estrus cycle were given PHA intravaginally.Results:After treatment with low concentration(15.6-125μg/ml)of PHA,the duration of aggregation in the spermatozoa of different species was about 30 minutes,but the spermatozoa couldn't be killed.The aggregated sperm could survive for 7-12 hours at room temperature(22±7℃).Treated with high concentration(≥250μg/ml)of PHA,the spermatozoa of different species were shocked and then died.The PHA administrated vaginally had good effects on protecting against pregnancy.Conclusions:The PHAextracted from white kidney bean native to Yunnan can aggregate the sperm membrane proteins in different species.It maybe used as a potential con-traception and anti-early pregnancy agent.