基于VEGF/VEGFR-2和Ang-1/Ang-2/Tie-2信号通路探讨归肾丸合八珍汤对DOR大鼠卵巢的影响  

Investigating the effect of Guishen Pill(归肾丸)plus Bazhen Decoction(八珍汤)on the ovaries of DOR rats based on the VEGF/VEGFR-2 and Ang-1/Ang-2/Tie-2 signaling pathway

作　　者：高同庆 尹燕 金志春 胡红霞 曾婕 王珺 蔡紫璨 吴锋英 GAO Tong-qing;YIN Yan;JIN Zhi-chun;HU Hong-xia;ZENG Jie;WANG Jun;CAI Zi-can;WU Feng-ying(Hubei University of Chinese Medicine,Wuhan 430065,China;Maternal and Child Health Hospital of Hubei Province,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430070,China;Yangzhou Municipal Hospital of Traditional Chinese Medicine,Yangzhou 225100,China;Wuhan Institute of Technology,Wuhan 430205,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]华中科技大学同济医学院附属湖北妇幼保健院,湖北武汉430070 [3]扬州市中医院,江苏扬州225100 [4]武汉工程大学,湖北武汉430205

出　　处：《时珍国医国药》2025年第6期1029-1034,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金青年项目(22301228);全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)。

摘　　要：目的探讨归肾丸合八珍汤治疗卵巢储备功能减退(DOR)的可能机制。方法共选取60只SD大鼠,随机分为对照组(Control组)、环磷酰胺组(CTX组)、戊酸雌二醇组(EV组)、中药低、中、高剂量组(GSBZ-L/GSBZ-M/GSBZ-H组),除Control组外其余各组均建立DOR模型。用药21d后阴道脱落细胞学刘氏染色观察大鼠动情周期变化,ELISA法测定血清E_2、FSH和AMH的含量,HE和透射电镜分别观察卵巢组织的病理改变和超微结构,Western Blot法检测VEGF、VEGFR-2、Ang-1、Ang-2和Tie-2蛋白表达。结果与Control组比较,CTX组大鼠动情周期出现紊乱,卵巢超微结构发生改变,血清FSH水平升高(P<0.05)、AMH和E_2水平降低(P<0.05),EV组和GSBZ-L组血清E_2水平升高(P<0.05);大鼠卵巢组织中CTX组VEGF、EV组Ang-1和GSBZ-M组VEGFR-2蛋白水平均降低(P<0.05),GSBZ-H组VEGFR-2蛋白水平升高(P<0.05)。与CTX组比较,EV组和中药各组卵泡逐渐恢复正常生长,卵巢线粒体肿胀程度减轻、数量增多,血清FSH水平均有不同程度降低(P<0.05),EV组和中药各组大鼠血清AMH、E_2水平均升高(P<0.05);大鼠卵巢组织中EV组VEGF、GSBZ-M组卵巢Ang-1和Tie-2、GSBZ-H组VEGF、Ang-1和Tie-2蛋白水平均升高(P<0.05)。结论归肾丸合八珍汤可以改善DOR大鼠血清激素水平、卵巢结构,促进血管生成,其机制可能与上调VEGF/VEGFR-2和Ang-1/Ang-2/Tie-2信号通路有关。Objective To investigate the possible mechanism of Guishen Pill(归肾丸)combined with Bazhen Decoction(八珍汤),abbreviated as GSBZ,in treating diminished ovarian reserve(DOR).Methods Sixty SD rats were randomly assigned to the control group,cyclophosphamide(CTX)group,estradiol valerate(EV)group,and GSBZ low/medium/high-dose((GSBZ-L/GSBZ-M/GSBZ-H)groups.DOR model was established in all groups except the control group.After 21 days of treatment,vaginal exfoliative cytology with Liu's staining was used to observe the estrous cycle changes.Serum levels of E2,FSH and AMH were measured by ELISA.Ovarian pathological changes and ultrastructure were observed through HE staining and transmission electron microscopy(TEM)respectively.Western blot(WB)was used to detect VEGF,VEGFR-2,Ang-1,Ang-2 and Tie-2 protein expressions.Results Compared with the control group,the CTX group showed disrupted estrous cycles,altered ovarian ultrastructure,increased serum FSH levels(P<0.05),decreased AMH and E2 levels(P<0.05),while both EV and GSBZ-L groups showed increased E2 levels(P<0.05).Ovarian VEGF in the CTX group,Ang-1 in the EV group,and VEGFR-2 protein levels in the GSBZ-M group were decreased(P<0.05),while VEGFR-2 protein levels in GSBZ-H group was increased(P<0.05).Compared with CTX group,EV and GSBZ groups exhibited recovered follicular growth,reduced ovarian mitochondrial swelling with increased quantity,decreased serum FSH levels(P<0.05)in varied degrees,and increased serum AMH and E2 levels(P<0.05).Ovarian VEGF in the EV group,Ang-1 and Tie-2 in the GSBZ-M group,as well as VEGF,Ang-1 and Tie-2 in the GSBZ-H group were all increased(P<0.05).Conclusion GSBZ can improve serum hormone levels and ovarian structure in DOR rats while promoting angiogenesis,and its mechanism may be associated with the up-regulation of the VEGF/VEGFR-2 and Ang-1/Ang-2/Tie-2 signaling pathways.

关 键 词：卵巢储备功能减退 归肾丸 八珍汤 血管生成 精血理论 

分 类 号：R285.5[医药卫生—中药学]