补肾调经方及逍遥丸对促性腺激素预处理小鼠排卵关键因子HIF-1α和EDN2的影响  

Effects of Bushen Tiaojing Recipe and Xiaoyao Pill on ovulation key factor HIF-1αand EDN2 of gonadotropin pretreatment mice

作　　者：宋翠淼[1,2,3] 段彦苍 贺明[1,2,3] 徐华洲 杜惠兰[1,2,3] SONG Cui-miao;DUAN Yan-cang;HE Ming;XU Hua-zhou;DU Hui-lan(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Collaborative Innovation Center of Integrated Chinese and Western Medicine on Reproductive Disease,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrated Chinese and Western Medicine on Liver-kidney Patterns,Shijiazhuang 050200,China)

机构地区：[1]河北中医学院,石家庄050200 [2]中西医结合生殖疾病协同创新中心,石家庄050200 [3]河北省中西医结合肝肾病证重点实验室,石家庄050200

出　　处：《中华中医药杂志》2020年第4期1714-1718,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81303002);河北省自然科学基金项目(No.H2013206117);河北省教育厅项目(No.ZD2015013)。

摘　　要：目的:观察补肾调经方、逍遥丸对性未成熟超促排卵小鼠卵巢缺氧诱导因子^-1α(HI F^-1α)及其下游因子内皮素2(E D N2)的影响,探讨补肾调经方、逍遥丸诱发排卵的作用机制。方法:雌性未成熟昆明小鼠随机分为对照组、补肾组和疏肝组,每组36只。补肾组、疏肝组小鼠先分别灌服补肾调经Ⅱ号方18.70g·kg^-1·d^-1、逍遥丸高剂量2.34g·kg^-1·d^-1各3天促进卵泡发育,再分别灌服补肾调经Ⅲ号方28.57g·kg^-1·d^-1、逍遥丸低剂量1.17g·kg^-1·d^-1各2天以诱发排卵。实验第3天,3组小鼠均腹腔注射用血促性素(PMSG)5U,48h后腹腔注射用绒促性素(hCG)5U。各组小鼠分别在hCG干预后0、8、12h各处死12只,采用RT-PCR、Western Blot和免疫组化方法检测卵巢组织HIF^-1α、EDN2 mRNA及其蛋白的表达。结果:hCG干预后0h,补肾组、疏肝组小鼠卵巢HIF^-1αmRNA及其蛋白表达较对照组显著增强(P<0.01),补肾组较疏肝组表达增强(P<0.05,P<0.01)。h CG干预后8h,补肾组、疏肝组卵巢HI F^-1α、E D N2 m R NA及其蛋白表达均较对照组显著增强(P<0.05,P<0.01),补肾组HIF^-1αmRNA及蛋白较疏肝组表达增强(P<0.01,P<0.05)。hCG干预后12h,补肾组、疏肝组卵巢HIF^-1α、EDN2 mRNA及其蛋白表达均较对照组显著增强(P<0.01,P<0.05),补肾组EDN2 mRNA较疏肝组表达增强(P<0.01)。结论:补肾调经方和逍遥丸可能通过促进卵泡发育,导致卵泡内产生局部缺氧环境,使HIF^-1αmRNA及其蛋白表达升高,进而促进其下游因子EDN2 mRNA及其蛋白表达,使卵泡顶端血管收缩、卵泡破裂而诱发排卵。Objective:To observe the effects of Bushen Tiaojing Recipe(BTR),Xiaoyao Pill(XYP)on superovulation of immature mice with ovarian hypoxia inducible factor^-1α(HIF^-1α)and its downstream factor of endothelin2(EDN2),and to study their mechanism of inducing ovulation.Methods:Immature Kunming mice were randomly divided into control group,Bushen group and Shugan group.The mice in the Bushen group and Shugan group were respectively fed with BTR II 18.70g·kg^-1·d^-1 or XYP high dose 2.34g·kg^-1·d^-1 for 3 days to promote follicular development,then they were respectively fed with BTR III 28.57g·kg^-1·d^-1 or XYP low dose of 1.17g·kg^-1·d^-1 for 2 days to induce ovulation.On the third day of the experiment,three groups of mice were injected intraperitoneally with PMSG 5U and injected hCG 5U intraperitoneally after 48 hours.Mice in each group were sacrificed at 0,8 and 12 hours after hCG intervention.RT-PCR,Western Blot,and immunohistochemistry methods were used to detect the mRNA and protein expressions of HIF^-1α,EDN2 in ovarian tissue.Results:After 0 hour of hCG intervention,the mRNA and protein expressions of HIF^-1αin the ovary of the Bushen and Shugan group were significantly higher than those in the control group(P<0.01),and the mRNA and protein expressions of HIF^-1αin of the Bushen group were significantly higher than those in Shugan group(P<0.05,P<0.01).After 8 hours of hCG intervention,the mRNA and protein expressions of HIF^-1αand EDN2 in the ovary of the Bushen and Shugan group were significantly higher than those in the control group(P<0.05,P<0.01),and the mRNA and protein expressions of HIF^-1αin the Bushen group were significantly higher than those in Shugan group(P<0.01,P<0.05).After 12 hours of hCG intervention,the mRNA and protein expressions of HIF^-1αand EDN2 in the ovary of the Bushen and Shugan group were significantly higher than those in control group(P<0.01,P<0.05),and the mRNA and protein expressions of EDN2 in the Bushen group were significantly higher than those in the Shugan gr

关 键 词：补肾调经方 逍遥丸 排卵 缺氧诱导因子-1Α 内皮素2 不孕 

分 类 号：R285.5[医药卫生—中药学]