连续超促排卵致肾精不足伴生育力低下雌性小鼠模型制备和比较研究  

Establishment and Comparative Study of Female Mice Models of Kidney Essence Deficiency with Low Fertility Induced by Continuous Ovarian Hyperstimulation

作　　者：刘奕[1] 陆华[1] 董文然 Liu Yi;Lu Hua;Dong Wenran(Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学,成都610075

出　　处：《世界科学技术-中医药现代化》2021年第5期1650-1657,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家科技部国家科技部重点研发计划(2018YFC1704305):不孕症“肾阳虚证”辩证标准的系统研究,负责人:陆华;国家中医药管理局中医药传承与创新“百千万”人才工程(岐黄工程),负责人:陆华。

摘　　要：目的探究应用连续超促排卵的方法,制备肾精不足伴生育力低下的雌性小鼠模型。比较连续超促排卵与反复超促排卵两种造模方式,探索最优造模方案。方法6周龄雌性昆明小鼠60只,随机分为自然周期组、超促排卵组、连续超促排卵15天组、连续超促排卵30天组、反复超促排卵5次组。其中,自然周期组于动情期腹腔注射绒毛膜促性腺激素(HCG)5IU,其余各组在小鼠动情间期予孕马血清促性腺激素(PMSG)5IU腹腔注射,48小时后腹腔注射HCG 5IU进行超促排卵。连续超促排卵15天即无间断连续超促排卵5个周期;连续超促排卵30天即无间断连续超促排卵10个周期。反复超促排卵5次组于每次注射HCG后间隔2天开始下1个周期,重复5个周期。最后1次注射HCG后解剖小鼠取卵,镜下计数卵细胞及分期;子宫组织进行HE染色,测定子宫内膜厚度、腺体情况;合笼计数产仔数及子代出生1周后存活情况。结果(1)连续超促排卵15天组,总获卵数降低(P<0.05),其中MII期卵数及占比均显著降低(P<0.01);产仔数降低(P<0.05)。连续超促排卵30天组,子宫腺体面积降低(P<0.05),腺体最大直径降低(P<0.05);MII期卵数及占比均降低(P<0.05),总获卵数显著降低(P<0.01);产仔数降低(P<0.05);出生1周子代存活数及存活率均显著降低(P<0.01)。(2)连续超促排卵15天组与反复超促排卵5次组比较,子宫内膜厚度、腺体情况、各级获卵数、子代数及存活率均无统计学差异。结论(1)连续超促排卵15天、连续超促排卵30天,均可以作为肾精不足伴生育力低下的研究模型。(2)连续超促排卵15天是时间最优化的造模方案。Objective To explore the method of continuous ovarian hyperstimulation to prepare the female mouse model of kidney essence deficiency with low fertility,and to compare the two models of continuous and repeated ovarian hyperstimulation,and to explore the optimal model.Methods Sixty 6-week-old female Kunming mice were randomly divided into natural cycle group,ovarian hyperstimulation group,15 day continuous ovarian hyperstimulation group,30 day continuous ovarian hyperstimulation group and 5 times repeated ovarian hyperstimulation group.In the natural cycle group,hCG 5 IU was injected intraperitoneally during the estrous period,and PMSG 5 IU was injected intraperitoneally in the other groups during the inter estrous period.48 hours later,hCG 5 IU was injected intraperitoneally for ovarian hyperstimulation.Continuous ovarian hyperstimulation for 15 days was 5 cycles of continuous ovarian hyperstimulation,and continuous ovarian hyperstimulation for 30 days is 10 cycles of continuous ovarian hyperstimulation.In the group of5 cycles of repeated ovarian hyperstimulation,the next cycle started 2 days after each injection of hCG,and repeated 5 cycles.After the last injection of hCG,the mice were dissected and the oocytes were collected,and the oocytes and stages were counted under the microscope;the endometrial thickness and glands were measured by HE staining of uterine tissue;the litter size was counted in cages and the survival of the offspring one week after birth.Results(1)In the 15 day group,the total number of retrieved eggs decreased(P<0.05),the number and proportion of MII stage eggs decreased significantly(P<0.01),and the number of litter decreased(P<0.05).In the 30 day group,the area of uterine gland and the maximum diameter of gland were significantly decreased(P<0.05),the number and proportion of MII stage eggs were significantly decreased(P<0.05),the total number of retrieved eggs was significantly decreased(P<0.01),the number of litter was significantly decreased(P<0.05),and the survival number and survival

关 键 词：肾精不足 生育力低下 小鼠模型 连续超促排卵 

分 类 号：R2-03[医药卫生—中医学]