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刘英莲, 王献, 张文龙, 岳雯, 周夏芝.加减养精种玉汤通过Rictor/mTORC2通路改善大鼠卵巢储备功能的机制研究[J].湖南中医药大学学报英文版,2024,44(8):1360-1365.[Click to copy
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| 加减养精种玉汤通过Rictor/mTORC2通路改善大鼠卵巢储备功能的机制研究 |
| 刘英莲,王献,张文龙,岳雯,周夏芝 |
| (海南医科大学中医学院, 海南 海口 571199) |
| 摘要: |
| 目的 探讨加减养精种玉汤通过雷帕霉素不敏感性伴随蛋白（rapamycin-insensitive companion of mTOR, Rictor）/哺乳动物雷帕霉素靶蛋白复合物2（mammalian targeting of rapamycin complex 2, mTORC2）通路改善大鼠卵巢储备功能的作用机制。方法 准备动情周期正常的雌性SD大鼠32只，随机选取其中8只作为空白组（等剂量生理盐水灌胃），其余SD大鼠首次腹腔注射环磷酰胺50 mg/mL后连续14 d腹腔注射8 mg/kg诱导卵巢储备功能减退（diminished ovarian reserve, DOR）模型。按随机数字表法将造模成功的24只大鼠分为模型组（等剂量生理盐水灌胃）、中药组（加减养精种玉汤3.39 g/kg灌胃）、卵泡刺激素（follicle stimulating hormone, FSH）组（FSH 0.105 mg/kg灌胃）。记录大鼠动情周期；HE染色观察左侧卵巢组织病理学变化并计算各组大鼠左侧卵巢窦卵泡数量；计算子宫、左侧卵巢系数；ELISA法测定FSH、黄体生成素（luteinizing hormone, LH）、雌二醇（estradiol, E2）、抗米勒管激素（anti-Müllerian hormone, AMH）水平；Western blot检测大鼠左侧卵巢组织Rictor、mTORC2蛋白表达水平。结果 与空白组相比，模型组动情周期次数减少（P＜0.05）;左侧卵巢成熟卵泡少;左侧卵巢指数、子宫指数、左侧卵巢窦卵泡数量均降低（P＜0.05，P＜0.01）；血清AMH、E2降低（P＜0.01），LH、FSH升高（P＜0.01）；左侧卵巢组织Rictor、mTORC2蛋白相对表达量升高（P＜0.01）。与模型组比，中药组及FSH组左侧卵巢结构改善，各级卵泡数量增加；左侧卵巢指数、子宫指数、左侧卵巢窦卵泡数量均上升（P＜0.05）；血清AMH、E2升高（P＜0.01），LH、FSH降低（P＜0.01）；左侧卵巢组织Rictor、mTORC2蛋白相对表达量降低（P＜0.01）。结论 加减养精种玉汤能够改善激素水平、调节卵巢卵泡细胞，从而起到改善DOR的作用，其机制可能与降低Rictor/mTORC2的表达有关。 |
| 关键词: 加减养精种玉汤 Rictor/mTORC2通路 卵巢储备功能 动情周期 环磷酰胺诱导 |
| DOI：10.3969/j.issn.1674-070X.2024.08.002 |
| Received:March 23, 2024 |
| 基金项目:海南省自然科学基金高层次人才项目（821RC565）；海南省卫生健康科技创新联合项目（WSJK2024MS164）。 |
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| Mechanism of modified Yangjing Zhongyu Decoction improving ovarian reserve function in rats through Rictor/mTORC2 pathway |
| LIU Yinglian, WANG Xian, ZHANG Wenlong, YUE Wen, ZHOU Xiazhi |
| (School of Chinese Medicine, Hainan Medical University, Haikou, Hainan 571199, China) |
| Abstract: |
| Objective To explore the effects of modified Yangjing Zhongyu Decoction (MYJZYD) on improving ovarian reserve function in rats through rapamycin-insensitive companion of mTOR (Rictor)/mammalian targeting of rapamycin complex 2 (mTORC2) pathway. Methods A total of 32 female SD rats with normal estrous cycle were included, eight of which were randomly selected as blank group (receiving equal dose of normal saline), and the diminished ovarian reserve (DOR) model was induced in the remaining rats through an initial intraperitoneal injection of cyclophosphamide at 50 mg/ml followed by continuous intraperitoneal injections 8 mg/kg per day for 14 days. According to random number table method, 24 rats were divided into model group (receiving equal dose of normal saline), Chinese medicine group (receiving MYJZYD 3.39 g/kg intragastric administration), and follicle-stimulating hormone (FSH) group (receiving FSH 0.105 mg/kg intragastric administration). The estrous cycle of rats was recorded, and the histopathological changes in the left ovary were observed by HE staining. The number of antral follicles in the left ovary of rats in each group was calculated, and the coefficients of uterus and the left ovary were calculated. Serum levels of FSH, luteinizing hormone (LH), estradiol (E2), and anti-Müllerian hormone (AMH) were measured by ELISA. The expression levels of Rictor and mTORC2 proteins in the left ovarian tissue of rats were determined by Western blot. Results Compared with the blank group, the model group showed a reduction in the number of estrous cycles (P＜0.05); the left ovarian structure was relatively disordered with fewer mature follicles; the left ovarian index, uterine index, and the number of antral follicles in the left ovary were all decreased (P＜0.05, P＜0.01), the serum AMH and E2 levels decreased (P＜0.01), while serum LH and FSH levels increased (P＜0.01). The relative expression levels of Rictor and mTORC2 proteins in the left ovarian tissue were higher (P＜0.01). Compared with the model group, the structure of the left ovary was improved and the number of follicles at all levels was significantly lower in the Chinese medicine group and FSH group. The left ovarian index, uterine index, and the number of antral follicles in the left ovary all increased (P＜0.05). Serum AMH and E2 levels increased (P＜0.01), while LH and FSH levels decreased (P＜0.01); the relative expression levels of Rictor and mTORC2 proteins in the left ovarian tissue were higher (P＜0.01). Conclusion MYJZYD can improve hormone levels, regulate ovarian follicle cells, thus playing a role in improving DOR. Its mechanism may be related to reducing the expression of Rictor/mTORC2. |
| Key words: modified Yangjing Zhongyu Decoction Rictor/mTORC2 pathway ovarian reserve function estrous cycle cyclophosphamide induction |
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