| 何仙春,屠雯妍,叶利群,林启笛,殷一红,沈颖,张晓芳.滋肾活血方对卵巢储备功能减退大鼠原代颗粒细胞的作用及机制研究[J].浙江中西医结合杂志,2025,35(9): |
| 滋肾活血方对卵巢储备功能减退大鼠原代颗粒细胞的作用及机制研究 |
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| 投稿时间：2025-01-21 修订日期：2025-07-29 |
| DOI： |
| 中文关键词: 滋肾活血方 卵巢储备功能减退 氧化应激 Nrf2/HO-1信号通路 |
| 英文关键词:Kidney-nourishing and blood-activating prescription Diminished ovarian reserve Oxidative stress Nrf2/HO-1 signaling pathway |
| 基金项目:国家中医药管理局科技司-浙江省中医药管理局共建科技计划项目(No.GZY-ZJ-KJ-23090);宁波市科技计划项目(No.2022J285)第一作者：何仙春(1999-),女,2022级浙江中医药大学在读硕士；汉族；籍贯：海南省陵水黎族自治区；研究方向：多囊卵巢综合征、宫颈疾病、不孕不育等妇科内分泌疾病临床研究,E-mail:a17860506959@163.com ,屠雯妍1 ,叶利群 通讯作者：叶利群(1973-),女,医学硕士,宁波市中医院妇二科主任医师,硕士研究生导师,浙江中医药大学教授；汉族；籍贯：浙江省宁波市；研究方向：多囊卵巢综合征、宫颈疾病、不孕不育等妇科内分泌疾病临床研究,E-mail:yeliqunnb@sina.com *,林启笛3 ,殷一红4 ,沈颖5 ,张晓芳6 |
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| 中文摘要: |
| 目的：探讨滋肾活血方调控Nrf2/HO-1信号通路改善卵巢储备功能减退(DOR)大鼠颗粒细胞氧化应激损伤的作用机制。方法 ：筛选36只SD雌性大鼠随机分为正常组,模型组,滋肾活血方(低、中、高)剂量组,阳性药组(戊酸雌二醇),除正常组外各组大鼠灌胃给予雷公藤多苷(30mg/kg/d)2周建立DOR模型；滋肾活血方组分别以中药汤剂低(7.5g/kg)、中(15g/kg)、高(30g/kg)剂量灌胃,阳性药组予戊酸雌二醇连续2周灌胃,取血和测定大鼠卵巢指数和体质量、卵巢组织形态、血清FSH、LH、AMH的浓度；再次筛选32只雌性大鼠进行分组和按照按照上述造模方法进行造模,DOR模型大鼠造模成功后分离正常组和模型组大鼠卵巢颗粒细胞,用15只SD雌性大鼠所制备滋肾活血方含药血清(低、高)进行处理,用于测定细胞内谷胱甘肽 (GSH)、过氧化氢酶(CAT)和丙二醛(MDA)含量,核因子E2相关因子2(Nrf2)和诱导型一氧化氮合酶(iNOS)、血红素氧合酶1(HO-1)的mRNA和蛋白表达水平。结果：与正常组比较,模型组大鼠动情周期紊乱,卵巢呈功能减退样改变；体质量、卵巢指数和血清AMH、颗粒细胞中CAT和GSH含量、Nrf2、HO-1 mRNA及蛋白表达显著下降(p<0.05,p<0.01),FSH、MDA、iNOSmRNA及蛋白表达水平显著升高(p<0.05,p<0.01,p<0.001)；与模型组比较,阳性药和滋肾活血方三个剂量组卵巢内各级卵泡发育数量增多,动情周期均有恢复,卵巢指数、AMH升高(p<0.05,p<0.01,p<0.001),滋肾活血方高剂量组与阳性药组FSH水平明显下降(p<0.05,p<0.01)；滋肾活血方高剂量组颗粒细胞内GSH、CAT活性升高,而MDA含量明显下降(p<0.05)；Nrf2 、HO-1mRNA和蛋白水平明显升高,滋肾活血方低、高剂量组iNOS蛋白表达水平显著下降(p<0.01,p<0.001)。结论：滋肾活血方能够通过调控Nrf2/HO-1信号通路的表达,提高卵巢颗粒细胞抗氧化能力,改善卵巢储备功能。 |
| 英文摘要: |
| Objective: To explore the mechanism by which Kidney-nourishing and blood-activating prescription regulates the Nrf2/HO-1 signaling pathway to improve oxidative stress injury in granulosa cells of rats with diminished ovarian reserve (DOR). Methods: Thirty-six SD female rats were randomly divided into normal group, model group, Kidney-nourishing and blood-activating prescription (low, medium, high) dose groups, and positive drug group (estradiol valerate). Except for the normal group, rats in each group were intragastrically administered with tripterygium glycosides (30 mg/kg/d) for 2 weeks to establish the DOR model. The Kidney-nourishing and blood-activating prescription groups were intragastrically administered with the decoction at low (7.5 g/kg), medium (15 g/kg), and high (30 g/kg) doses, and the positive drug group was intragastrically administered with estradiol valerate for 2 consecutive weeks. Blood was collected, and the ovarian index, body weight, ovarian tissue morphology, and serum concentrations of FSH, LH, and AMH were measured. Another 32 female SD rats were selected and grouped, and the DOR model was established using the same method. After successful establishment of the DOR model, granulosa cells from the ovaries of normal and model group rats were isolated. The granulosa cells were treated with the drug-containing serum of Kidney-nourishing and blood-activating prescription (low and high doses) prepared from 15 SD female rats to measure the intracellular levels of glutathione (GSH), catalase (CAT), and malondialdehyde (MDA), as well as the mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), inducible nitric oxide synthase (iNOS), and heme oxygenase 1 (HO-1). Results: Compared with the normal group, the model group rats showed disrupted estrous cycles and ovarian atrophy; body weight, ovarian index, and serum AMH levels, as well as CAT and GSH contents in granulosa cells, and Nrf2 and HO-1 mRNA and protein expression levels were significantly decreased (p < 0.05, p < 0.01), while FSH, MDA, iNOS mRNA and protein expression levels were significantly increased (p < 0.05, p < 0.01, p < 0.001). Compared with the model group, the positive drug and Kidney-nourishing and blood-activating prescription groups showed increased numbers of follicles at all stages in the ovaries, restored estrous cycles, increased ovarian index and AMH levels (p < 0.05, p < 0.01, p < 0.001), and significantly decreased FSH levels in the high-dose Kidney-nourishing and blood-activating prescription group and the positive drug group (p < 0.05, p < 0.01). The high-dose Kidney-nourishing and blood-activating prescription group showed increased GSH and CAT activities and significantly decreased MDA content in granulosa cells (p < 0.05), as well as significantly increased Nrf2 and HO-1 mRNA and protein expression levels, and significantly decreased iNOS protein expression levels in the low and high-dose Kidney-nourishing and blood-activating prescription groups (p < 0.01, p < 0.001). Conclusion: Kidney-nourishing and blood-activating prescription can improve ovarian reserve function by regulating the expression of the Nrf2/HO-1 signaling pathway and enhancing the antioxidant capacity of ovarian granulosa cells. |
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