温阳化浊方对高龄模型小鼠生殖衰老、卵巢机械微环境及子代生殖潜能的影响  

Effect of Wenyang Huazhuo Formula(温阳化浊方)on Reproductive Aging,Ovarian Mechanical Microenvironment,and Offspring Reproductive Potential in Aged Model Mice

作　　者：徐佳淇 赵晓丽 江楠 李凯茜 丁亚飞 温子木 贾滢瑛 姜梦君 夏天[1,2] XU Jiaqi;ZHAO Xiaoli;JIANG Nan;LI Kaixi;DING Yafei;WEN Zimu;JIA Yingying;JIANGMengjun;XIA Tian(First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin,300193;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion;Tianjin University of Traditional Chinese Medicine)

机构地区：[1]天津中医药大学第一附属医院,天津市西青区300193 [2]国家中医针灸临床医学研究中心 [3]天津中医药大学

出　　处：《中医杂志》2025年第6期612-620,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82274570)。

摘　　要：目的从卵巢机械微环境角度探讨温阳化浊方改善生殖衰老的可能作用机制。方法实验设5个组:20只3月龄雌性小鼠设为3月龄组,20只6月龄雌性小鼠设为6月龄组,20只5月龄雌性小鼠设为6月龄+温阳化浊方组,20只9月龄雌性小鼠设为9月龄组,20只8月龄雌性小鼠设为9月龄+温阳化浊方组。6月龄+温阳化浊方组、9月龄+温阳化浊方组小鼠给予温阳化浊方41.2 g/(kg·d)灌胃,每日1次,连续4周。其余三组不给予任何干预。ELISA方法检测生殖激素水平;HE染色计数各级卵泡数量;测定卵巢组织透明质酸(HA)含量和胶原纤维含量,评价卵巢机械微环境;采用超数促排法观察各组小鼠获卵数,以及产生的子代数量和出生体质量情况评价生育力;观察各组小鼠子代雌性个体的卵母细胞体外受精和囊胚培养情况,评价温阳化浊方对子代生殖潜能的影响。结果与3月龄组比较,6月龄组、9月龄组小鼠血清促性腺激素释放激素(GnRH)、促卵泡素(FSH)和黄体生成素(LH)水平,卵巢胶原纤维含量,始基卵泡和次级卵泡数量,获卵数、子代数量显著减少,初级卵泡、窦卵泡和闭锁卵泡数量均升高,抗缪勒管激素(AMH)、卵巢基质HA含量及子代小鼠体外培养的卵母细胞受精率、卵裂率及囊胚形成率降低(P<0.05)。与6月龄组比较,6月龄+温阳化浊方组小鼠血清GnRH、FSH和LH水平,初级卵泡、窦卵泡和闭锁卵泡数量减均显著少,AMH、卵巢基质HA含量,子代小鼠卵母细胞的受精率,始基卵泡和次级卵泡数量,获卵数和子代出生体质量增加(P<0.05)。与9月龄组相比,9月龄+温阳化浊方组小鼠GnRH、FSH、胶原纤维含量,初级卵泡、窦卵泡和闭锁卵泡数量均减少,AMH、卵巢基质HA含量,始基卵泡和次级卵泡数量,获卵数、子代数量、出生体质量,子代小鼠卵母细胞的受精率、卵裂率及囊胚形成率均增加(P<0.05)。结论温阳化浊方可明显改善生殖中期及晚Objective To explore the possible mechanisms of Wenyang Huazhuo Formula(温阳化浊方,WHF)in improving reproductive aging from the perspective of the ovarian mechanical microenvironment.Methods The experiment included five groups,3-month group(20 female mice at 3 months of age),6-month group(20 female mice at 6 months of age),6-month+WHF group(20 female mice at 5 months of age treated with WHF),9-month group(20 female mice at 9 months of age),and 9-month+WHF group(20 female mice at 8 months of age treated with WHF).The 6-month+WHF group and 9-month+WHF group were orally administered WHF 41.2 g(/kg·d)once daily for 4 consecutive weeks.The other three groups received no intervention.Reproductive hormone levels were measured by ELISA.HE staining was used to count the numbers of various stages of follicles.Ovarian hyaluronic acid(HA)content and collagen fiber content were measured to evaluate the ovarian mechanical microenvironment.Superovulation was performed to observe the number of eggs obtained,as well as the number of offspring and birth weight to assess fertility.The in vitro fertilization and blastocyst culture of oocytes from female offspring in each group were observed to evaluate the effect of WHF on offspring reproductive potential.Results Compared with the 3-month group,the 6-month group and 9-month group showed significantly decreased serum levels of gonadotropin-releasing hormone(GnRH),follicle-stimulating hormone(FSH),and luteinizing hormone(LH),decreased ovarian collagen content,and reduced numbers of primordial and secondary follicles.In contrast,the numbers of primary follicles,antral follicles,and atretic follicles increased.The levels of anti-Müllerian hormone(AMH),ovarian HA content,and the fertilization rate,cleavage rate,and blastocyst formation rate of oocytes from offspring were significantly lower(P<0.05).Compared with the 6-month group,the 6-month+WHF group showed significantly reduced serum levels of GnRH,FSH,and LH,with a significant decrease in primary follicles,antral follicles,and

关 键 词：生殖衰老 卵巢机械微环境 温阳化浊方 卵巢储备力 生殖激素 生殖潜能 

分 类 号：R71[医药卫生—妇产科学]