摘要
目的 研究雷公藤内酯酮的雄性抗生育作用机制。方法 用雷公藤内酯酮 2 0 0μg/ (kg· d)喂服Wistar大鼠 ,8周后取材。做大鼠睾丸精子计数、精子活动度、睾丸切片的形态学观察 ,精核蛋白提取以及基因 cyclinD1和 Cdk4m RNA的原位杂交实验。结果 实验组大鼠睾丸精子数量为 (2 6 .43± 3.90 )× 10 6 / g,比对照组大鼠的精子计数 (4 1.15± 5 .5 1)× 10 6 / g明显降低 (P<0 .0 1) ,精子活动度为 (6 6 .0± 6 .8) % ,较对照组 (88.0± 2 .0 ) %极显著降低 (P<0 .0 0 1) ;cyclin D1和 Cdk4基因表达在精子细胞中显著上升。结论 雷公藤内酯酮的雄性抗生育作用显著 ,作用机制可能主要是增加精子细胞 cyclin D1和 Cdk4基因的表达 ,抑制附睾精核蛋白的生物合成。
The male contraceptive mechanism in rats of triptonide,isolated from Multiglycosides of Tripterygium wilfordii(GTW),was investigated. Methods The dosage of triptonide was 200 μg/(kg·d),fed for 8 weeks via gastric intubation. Sperm counts,sperm motility, and sperm silver staining were used to test the antispermatogenic effect of triptonide;the study of morphology was learned from paraffin sections of testis and visceras of triptonide-treated rats;protamine was abstracted from testis by electrophoresis;using antisense mRNA of cyclin D1 and Cdk4 as prodes,we examed the expression of cyclin D1 and Cdk4 through hybridization in situ. Results Sperm counts (26.43±3.90)×10 6/g and sperm motility(66.0±6.8)% of triptonide-treated rats were substantially different from those of control,(41.15±5.51)×10 6/g(P<0.01) and (88.0±2.0)% (P<0.001).Paraffin sections of testis of triptonide-treated rats stained by PAS showed that the number of spermatids per 100 sertoli cells (9.15±0.33) was remarkably decreased compared with control’s (10.73±0.38) (P<0.001).At the same time,delaying sperm-released process was observed,which affects the nature course of sperm.And the extended vacuoles of vary sizes,the segregation of cells in lumens were also found in the cross-section of testis of rats treated with triptonide.In addition,the contents of protamine (4.6±2.2)% was extremely decreased,compared with the control’s (40.6±7.2)% (P<0.001).By using antisense mRNA of cyclin D1 and Cdk4 as prodes,we detected that the expression of the genes was notably increased in spermatids. Conclusions The results suggest the antispermatogenic effect of triptonide is very well and it mainly plays its role on spermatids by increasing the expression of cyclin D1 and Cdk4,meanwhile,decreasing the biosynthesis of protamine.The appropriate target region of triptonide makes itself most potency to be a suitable male antifertility drug.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2000年第3期224-226,共3页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金 !(396 70 76 3)
卫生部科学研究基金 !(96 -1-0 74)
计生委八五攻关项目! (85 -918)资助
