Li Meifang, Xie Li, Li Yang, Liu Jian, Nie Guangning, Yang Hongyan
Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006 China.
Department of Gynaecology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510120 China.
Chin Med. 2020 Aug 5;15:83. doi: 10.1186/s13020-020-00362-6. eCollection 2020.
Huyang Yangkun Formula (HYYKF) was developed based on theory of traditional Chinese medicine as well as clinical experience and used to improve ovarian function of premature ovarian insufficiency (POI) patients. Transplantation of embryonic stem cells (ESCs) has great potential in improving POI, and studies have confirmed that traditional Chinese medicine promoted the treatment effect of ESCs. In the present study, we compared the effect of combining HYYKF and ESCs, single HYYKF treatment and single ESCs intervention on POI mice to explore the effect of combination of HYYKF and ESCs in improving ovarian function.
C57BL/6 mice were used to create a POI model by 15-day intraperitoneal injection of 160 mg/kg of 4-vinylcyclonhexene diepoxide (VCD) and then treated with HYYKF, ESCs transplantation and combination of ESCs and HYYKF. When the treatments were finished, estrus cycle, ovarian follicle counting, serum sex hormone level, and expression of key nodes in the transforming growth factor beta/transforming growth factor beta-activated kinase 1 (TGF-β/TAK1) signaling pathway were determined.
Combination therapy brought down the abnormal estrus cycle rate to 5.26%, significantly lower than that of HYYKF or ESCs alone (30%, 25%, respectively). The numbers of follicles at all levels were increased significantly in the combination ESCs with HYYKF group (P < 0.05), especially that of antral follicles (P < 0.01), which was not increased significantly when HYYKF or ESCs was single used. The level of anti-Mullerian hormone (AMH) was more significantly increased in the combination ESCs with HYYKF group (P < 0.01) than that of HYYKF or ESCs alone (both P < 0.05). The expression of the key nodes TGF-β1, TAK1, JNK, Smad4 and FSHR in the TGF-β/TAK1 pathway were obviously affected in the SCHY group.
Both HYYKF and ESCs improve the ovarian function of POI induced by VCD, and a combination of HYYKF and ESCs has the advantage that they work together to promote follicles developing probably by inhibiting expression of the TGF-β1/TAK1 pathway.
胡杨养坤方（HYYKF）是基于中医理论及临床经验研制而成，用于改善卵巢早衰（POI）患者的卵巢功能。胚胎干细胞（ESCs）移植在改善POI方面具有巨大潜力，且研究证实中药可提高ESCs的治疗效果。在本研究中，我们比较了HYYKF与ESCs联合、单独使用HYYKF治疗以及单独ESCs干预对POI小鼠的影响，以探究HYYKF与ESCs联合改善卵巢功能的效果。
采用C57BL/6小鼠，通过腹腔注射160mg/kg的4-乙烯基环己烯二环氧化物（VCD）15天建立POI模型，然后分别用HYYKF、ESCs移植以及ESCs与HYYKF联合进行治疗。治疗结束后，测定发情周期、卵巢卵泡计数、血清性激素水平以及转化生长因子β/转化生长因子β激活激酶1（TGF-β/TAK1）信号通路关键节点的表达。
联合治疗使异常发情周期率降至5.26%，显著低于单独使用HYYKF或ESCs组（分别为30%、25%）。ESCs与HYYKF联合组各级卵泡数量均显著增加（P＜0.05），尤其是窦卵泡数量（P＜0.01），单独使用HYYKF或ESCs时窦卵泡数量未显著增加。ESCs与HYYKF联合组抗苗勒管激素（AMH）水平升高幅度比单独使用HYYKF或ESCs组更显著（P＜0.01，单独使用HYYKF或ESCs组P均＜0.05）。TGF-β/TAK1通路中关键节点TGF-β1、TAK1、JNK、Smad4和FSHR的表达在联合组中受到明显影响。
HYYKF和ESCs均可改善VCD诱导的POI小鼠的卵巢功能，HYYKF与ESCs联合具有协同促进卵泡发育的优势，可能是通过抑制TGF-β1/TAK1通路的表达实现的。
