CAJ | 학술논문

目的：探讨雷公藤甲素诱导NIH小鼠卵巢损伤的作用及机制。方法30只雌性NIH小鼠随机分为3组：正常对照组、雷公藤甲素低剂量组及雷公藤甲素高剂量组，分别以含5％DMSO的生理盐水和25、50μg/kg雷公藤甲素连续灌胃50 d。第41天起，行连续阴道涂片。第47天，行超排卵实验。于第50天处死小鼠，收集计数输卵管中卵子，留取卵巢行常规病理染色、免疫组织化学和TUNEL检测等。阴道涂片行巴氏染色检测小鼠性动周期，HE染色检测小鼠生殖器官病理，计数各级卵泡，TUNEL染色检测小鼠卵巢颗粒细胞凋亡，免疫组织化学法检测小鼠卵巢颗粒细胞细胞色素C（Cyt C）、凋亡蛋白酶活化因子-1（APAF-1）、X连锁凋亡抑制蛋白（XIAP）等的表达变化。结果雷公藤甲素低剂量组和雷公藤甲素高剂量组NIH小鼠卵巢均可见显著损伤，表现为：与正常对照组比较，雷公藤甲素低剂量组和雷公藤甲素高剂量组超排卵子质量低下；卵子数量显著减少；各级进展卵泡丢失，闭锁卵泡增加；卵巢颗粒细胞凋亡增多，卵泡内颗粒细胞线粒体凋亡相关蛋白Cyt C、APAF-1表达上调、而抗凋亡蛋白XIAP表达下调，差异有统计学意义（ P＜0.05）。结论雷公藤甲素可能通过诱导NIH小鼠卵巢颗粒细胞线粒体凋亡发挥卵巢损伤作用。高、低剂量雷公藤甲素对NIH小鼠卵巢损伤差异不大。
목적：탐토뢰공등갑소유도NIH소서란소손상적작용급궤제。방법30지자성NIH소서수궤분위3조：정상대조조、뢰공등갑소저제량조급뢰공등갑소고제량조，분별이함5％DMSO적생리염수화25、50μg/kg뢰공등갑소련속관위50 d。제41천기，행련속음도도편。제47천，행초배란실험。우제50천처사소서，수집계수수란관중란자，류취란소행상규병리염색、면역조직화학화TUNEL검측등。음도도편행파씨염색검측소서성동주기，HE염색검측소서생식기관병리，계수각급란포，TUNEL염색검측소서란소과립세포조망，면역조직화학법검측소서란소과립세포세포색소C（Cyt C）、조망단백매활화인자-1（APAF-1）、X련쇄조망억제단백（XIAP）등적표체변화。결과뢰공등갑소저제량조화뢰공등갑소고제량조NIH소서란소균가견현저손상，표현위：여정상대조조비교，뢰공등갑소저제량조화뢰공등갑소고제량조초배란자질량저하；란자수량현저감소；각급진전란포주실，폐쇄란포증가；란소과립세포조망증다，란포내과립세포선립체조망상관단백Cyt C、APAF-1표체상조、이항조망단백XIAP표체하조，차이유통계학의의（ P＜0.05）。결론뢰공등갑소가능통과유도NIH소서란소과립세포선립체조망발휘란소손상작용。고、저제량뢰공등갑소대NIH소서란소손상차이불대。
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 .