王瓜根蛋白的柱层析分离及其抗生育作用

应用Ambelie.IRC.50柱层析分离纯化王瓜根抗早孕蛋白,图谱显示抗早孕蛋白达到基线分离,用SDS.PAGE不连续电泳证实是电泳纯。应用本工艺生产的王瓜根抗早孕蛋白作小白鼠抗着床试验,其ED_(100)(100％抑制率)为1.5mg/kg,是结晶天花粉ED_(100)(10mg/kg)的6倍以上,提示这种蛋白有可能作妇女房事后避孕的新药。对应用王瓜抗早孕蛋白注射后的小白鼠子宫作组织学鉴定,由显微组织切片显示王瓜根抗早孕蛋白较天花粉蛋白作用强,子宫组织发生了变化。用王瓜根做抗早孕蛋白口服试验,也证实其效果好。这提示王瓜根蛋白可制成简便口服避孕片。

Preparation study of the effect of Radix Trichosanthis cucumeroides proteins on terminating early pregnancy in mice

In China, some people have used a preciptaton method of actone to prepare Radix Trichosanthis proteins since 1973. We have, however developed a new method of preparative chromatography in Amberlite IRC-50 column. This Method revealed that it was high yield and performance. Its Chromatographic conditions are:Column Dimensions: 1x15cm; Bed Materials: Amberlite IRC-50; Mobile Phase: First Step-Distilled water; Second Step-0.06N Na_2 HPO_4+0.4N NaCI; Flow Rate: 1.0ml/10min; Sample vol 2ml Chart Speed: 1cm/20min. There are seven peaks after separated by column Amberlite-IRC-50. Some of these proteins were shown to terminate pregnancy in mice following subcutaneous adminis-tration on the 4~th and 6~ttdays of pregnancy. The ED_100 of peak 6 terminated early pregnancy in mice was 1.5mg/kg. It is much better than that of Radix Trichosanthis protein ED_100 10mg/kg which we have been used in clinic. Subcutaneous injection of Radix Trichosanthis cucumeroides protein at doses capable of terminating early pregnancy inhibited the decidual reaction in pseudopregnant mice. This inhibitory effect, as well as the effect to terminate early pregnancy increased with increasing dosage. Some of these proteins were also shown to terminate pregnancy in mice following oral oppication on the 6~th'and 8~thdays of pregnancy. As oral administration, theED_100 of terminating early pregnancy in mice was 30mg/kg. Base on these results, we considered that this protein should be used as contraceptive medicine for oral use, and that further intensive investigations should be carried out.