The role of granulose cell apoptosis in Triptolide-induced ovary injury in NIH mice

ZENG You-jia; SUN Hui-li; XU Yuan-zhao; HAN Peng-xun; LI Shun-min

Abstract：Objective To evaluate the role of Triptolide -induced ovarian injury in NIH mice .Methods The triptolide-induced mouse ovary injury model was constructed .30 healthy female NIH mice were randomly divided 3 groups:control group (5%DMSO in saline), low dose Triptolide group (25 μg/kg Triptolide in 5%DMSO saline) and high dose Triptolide group (50 μg/kg Triptolide in 5%DMSO saline).Intragastric administration was performed for 50 days.From Day 41 to Day 50, vagina smears were executed .Superovulation was performed after Day 47.On Day 50, the mice were executed , and all the ova were collected from oviducts and counted .Ovaries were also collected for pathological observation, immunohistochemical and TUNEL assessment .The estrous cycle was estimated by vaginal cytology method . HE staining was used for pathological observation of ovary , oviduct and uterus; and follicle count at different levels . TUNEL assay was used to detect the apoptotic granulose cell .Immunohistochemistry was performed to observe the expres-sion of Cyt C, APAF-1, XIAP and et al.in ovary tissue.Results Both low and high dose Triptolide induced significant ovary injury in NIH mice .Compared to control group , significantly less in ovum counts were observed in both Triptolide treatment groups after superovulation .Besides, the developed follicles loss and atresia follicles increase were also observed in the both Triptolide treatment groups , so were the reduction in glands and thinner muscle layer in oviducts and uteruses , and increased ovary apoptotic granulose cells in the both Triptolide groups .Mitochondrial apoptosis related proteins , inclu-ding Cyt C and APAF-1, were up-regulated;while anti-apoptosis protein XIAP was down -regulated in the both Trip-tolide groups .Conclusion Granulose cell apoptosis is observed in Triptolide -induced ovary injuries in NIH mice , which is probably via mitochondrial pathway .