Sun F, Wei W
Department of Pharmacy, General Hospital of Shaoxing Second Hospital, Shaoxing 312000, China.
Department of Obstetrics and Gynecology, Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine, Hangzhou 310000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Dec 20;41(12):1850-1856. doi: 10.12122/j.issn.1673-4254.2021.12.14.
To investigate the effect of combined treatment with gonadotropin-releasing hormone (GnRH) agonist and menstrual blood-derived stem cell (MenSC) transplantation on ovarian function in mice.
We tested the effect of GnRH agonist and MenSC transplantation, either alone or in combination, in a mouse model of premature ovarian failure (POF) induced by daily intraperitoneal injection of 120 mg/kg cyclophosphamide (CTX) for 15 days. Venirelin was administered at the daily dose of 0.1 mg via intravenous infusion, starting at 14 days before modeling; MenSC suspension (2× 106 cells) was injected into the ovarium 1 day after modeling. HE staining was used to observe ovarian pathologies of the mice. ELISA was used to detect serum levels of LH, FSH, AMH and E2. The expression levels of ki-67, Bcl-2, BAX, caspase 9, caspase 3, their cleavages, P-PI3K, PI3K, P-Akt and Akt proteins in ovarian granulosa cells were determined with Western blotting.
The mouse models of POF showed obvious ovarian fibrosis with increased atresia follicles and had significantly elevated serum levels of LH and FSH ( < 0.01), decreased serum levels of AMH and E2 ( < 0.01), significantly increased ovarian expression levels of caspase 9, caspase 3, and their cleavages ( < 0.01), and decreased expression levels of Ki-67, Bcl-2, P-PI3K and P-Akt ( < 0.01). The combined treatment with GnRH agonist and MenSC transplantation obviously inhibited the production of ovarian atresia follicles, lowered serum levels of LH and FSH ( < 0.01), and increased the levels of AMH and E2 ( < 0.05 or 0.01) in the mouse models. The combined treatment also inhibited the expressions of BAX and cleaved caspase 9 and caspase 3 proteins ( < 0.05 or 0.01) and enhanced the expressions of Ki-67, Bcl-2, P-PI3K, and p-Akt proteins ( < 0.05 or 0.01) in ovarian granulococcus cells.
In mouse models of POF, GnRH agonist combined with MenSC transplantation can reduce the generation of ovarian atresia and improve ovarian reserve by up-regulating PI3K-Akt signaling pathway to achieve ovarian repair and protection.
探讨促性腺激素释放激素（GnRH）激动剂与月经血源性干细胞（MenSC）移植联合治疗对小鼠卵巢功能的影响。
我们在通过每日腹腔注射120mg/kg环磷酰胺（CTX），持续15天诱导的小鼠卵巢早衰（POF）模型中，测试了GnRH激动剂和MenSC移植单独或联合使用的效果。从建模前14天开始，以每日0.1mg的剂量通过静脉输注给予戈那瑞林；建模后1天，将MenSC悬液（2×10⁶个细胞）注射到卵巢中。采用苏木精-伊红（HE）染色观察小鼠卵巢病理变化。采用酶联免疫吸附测定（ELISA）法检测血清促黄体生成素（LH）、促卵泡生成素（FSH）、抗缪勒管激素（AMH）和雌二醇（E₂）水平。用蛋白质免疫印迹法检测卵巢颗粒细胞中ki-67、Bcl-2、BAX、半胱天冬酶9（caspase 9）、半胱天冬酶3（caspase 3）及其裂解产物、磷酸化磷脂酰肌醇-3激酶（P-PI3K）、磷脂酰肌醇-3激酶（PI3K）、磷酸化蛋白激酶B（P-Akt）和蛋白激酶B（Akt）的表达水平。
POF小鼠模型显示卵巢明显纤维化，闭锁卵泡增多，血清LH和FSH水平显著升高（P<0.01），血清AMH和E₂水平降低（P<0.01），卵巢中caspase 9、caspase 3及其裂解产物的表达水平显著升高（P<0.01），而Ki-67、Bcl-2、P-PI3K和P-Akt的表达水平降低（P<0.01）。GnRH激动剂与MenSC移植联合治疗明显抑制了小鼠模型中卵巢闭锁卵泡的产生，降低了血清LH和FSH水平（P<0.01），并提高了AMH和E₂水平（P<0.05或0.01）。联合治疗还抑制了卵巢颗粒细胞中BAX以及裂解的caspase 9和caspase 3蛋白的表达（P<0.05或0.01），并增强了Ki-67、Bcl-2、P-PI3K和p-Akt蛋白的表达（P<0.05或0.01）。
在POF小鼠模型中，GnRH激动剂与MenSC移植联合应用可通过上调PI3K-Akt信号通路减少卵巢闭锁的发生，改善卵巢储备，从而实现卵巢的修复和保护。
