雷公藤甲素对模式生物雄性秀丽隐杆线虫的生殖毒性作用

Reproductive toxicity of Triptolide on male Caenorhabditis elegans,a model organism

目的研究中药雷公藤的主要成分之一雷公藤甲素的雄性生殖毒性及其作用机制。方法将L4期him-5雄虫在0.02、0.2、2.0 mg/L的雷公藤甲素中处理48 h。应用杂交试验观察雷公藤甲素对雄虫生育力的作用,通过观察精细胞形态发育、精细胞活化、精子运动能力,并应用实时荧光定量PCR检测精子发生相关基因的mRNA表达水平,探讨雷公藤甲素雄性生殖毒性的作用机制。结果雷公藤甲素处理48 h后,与溶剂对照组比较,2.0 mg/L雷公藤甲素组线虫后代数目显著减少,0.2、2.0 mg/L雷公藤甲素组线虫精细胞直径和截面积均显著降低,精子活化率显著降低、活动精子百分率显著下降。spe-10在0.02~2.0 mg/L雷公藤甲素组、spe-15在0.2、2.0 mg/L雷公藤甲素组、fer-1和folt-1在2.0 mg/L雷公藤甲素组均出现表达水平下调的现象。结论雷公藤甲素通过抑制him-5雄虫精子发生通路相关基因的表达水平,抑制精细胞形态发育和精子的形态和功能发育,降低him-5雄虫的生育力,提示精子发生损伤是雷公藤甲素发挥生殖毒性作用的重要途径之一。

AIM To study the reproductive toxicity of Triptolide,one of the main components of Tripterygium wilfordii Hook. f,on male Caenorhabditis elegans,and its potential mechanism. METHODS Expose L4 larva of him-5 nematodes to 48 h 0. 02,0. 2,2. 0 mg/L of Triptolide. The effect of Triptolide on the fertility of male C.elegans was assessed by crossing test,and the relevant mechanism was explored through measuring spermatids morphology development,spermatids activation,sperm motility and the relative mRNA expression levels of the related spermatogenesis genes. RESULTS Compared with the solvent control group,the 48 h Triptolide treatment induced significantly decreased brood size in 2. 0 mg/L Triptolide group,significant reduction in spermatid diameter,spermatid cross-sectional area,rates of spermatid activation,rates of motile sperm in 0. 2 and 2. 0 mg/L Triptolide groups,and decreased expression levels of spe-10 in 0. 02-2. 0 mg/L Triptolide groups,decreased spe-15 in0. 2-2. 0 mg/LTriptolide groups, and decreased fer-1 and folt-1 in 2. 0 mg/L Triptolide groups as well.CONCLUSION Triptolide decreases expression levels of the related genes in spermatogenesis,contributes to an array of indicators inhibition,the spermatids morphology development,the sperm morphology and function development,which leads to decreased fertility of male C. elegans. Therefore,spermatogenesis damage is one of the main pathway that Triptolide adversely affects the fertility.