| 丁秀丽,徐舜.葛根素介导自噬延缓环磷酰胺诱导卵巢早衰大鼠的作用研究[J].浙江中西医结合杂志,2025,35(7): |
| 葛根素介导自噬延缓环磷酰胺诱导卵巢早衰大鼠的作用研究 |
| Effect of puerarin mediated autophagy on delaying premature ovarian failure induced by cyclophosphamide in rats |
| 投稿时间：2024-11-29 修订日期：2025-05-16 |
| DOI： |
| 中文关键词: 葛根素 卵巢早衰 自噬 LC3-Ⅱ 环磷酰胺 |
| 英文关键词:puerarin Premature ovarian failure Autophagy LC3-Ⅱ cyclophosphamide |
| 基金项目:2023年衢州市市级科技攻关项目(2023K182) |
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| 中文摘要: |
| 目的 研究葛根素介导自噬对环磷酰胺诱导的卵巢早衰（Premature ovarian failure, POF）大鼠的影响。方法 6只SD大鼠为对照组，12只SD大鼠造模后随机分为POF组、葛根素组，各6只。采用环磷酰胺诱导大鼠构建POF模型，葛根素组给予腹腔注射50 mg/kg葛根素，对照组和POF组给予腹腔注射等量生理盐水，持续干预21 d。阴道涂片观察动情周期变化，酶联免疫吸附法检测血清中抗缪勒氏管激素（anti-Müllerian hormone, AMH）、雌二醇（Estradiol, E2）、促黄体生成素（Luteinizing hormone, LH）和促卵泡激素（Follicle-stimulating hormone, FSH）水平，苏木精-伊红染色观察卵巢组织病理变化，透射电镜检测卵巢组织中自噬体数量变化，免疫组化测定微管相关蛋白1轻链3-Ⅱ（microtubule associated protein 1 light chain 3-Ⅱ, LC3-Ⅱ）表达水平，蛋白免疫印迹检测苄氯素1（Beclin 1）、自噬衔接蛋白62（autophagy adapter protein 62, p62）的表达。结果 与对照组比较，环磷酰胺诱导后，POF组大鼠动情周期紊乱，卵巢组织体积萎缩，卵泡数量减少，AMH[（491.50±47.77）pg/mL比（1228.63±100.66）pg/mL, P<0.01]、E2[（32.63±2.01）pg/mL比（54.65±7.37）pg/mL, P<0.01]含量显著降低，LH[（18.43±1.16）mIU/mL比（10.08±0.27）mIU/mL, P<0.01]、FSH[（12.50±1.41）IU/L比（7.55±0.86）IU/L, P<0.01]含量显著升高。与POF组比较，葛根素组大鼠动情周期恢复正常，卵巢结构恢复，卵泡数量增加，AMH[（859.57±159.89）pg/mL比（491.50±47.77）pg/mL, P<0.01]、E2[（48.86±6.60）pg/mL比（32.63±2.01）pg/mL, P<0.01]含量显著降低，LH[（12.43±1.70）mIU/mL比（18.43±1.16）mIU/mL, P<0.01]、FSH[（7.48±0.56）IU/L比（12.50±1.41）IU/L, P<0.01]含量显著升高。此外，与POF组比较，葛根素能促进自噬小体生成，促进LC3-Ⅱ的表达[（0.19±0.03）比（0.11±0.03）, P<0.05]，上调Beclin 1[（0.919±0.044）比（0.395±0.048）, P<0.01]和下调p62/GAPDH[（0.851±0.063）比（1.629±0.105）, P<0.01]蛋白的表达。结论 葛根素通过促进自噬减轻环磷酰胺诱导卵巢早衰损伤。 |
| 英文摘要: |
| Objective To study the effect of puerarin mediated autophagy on Premature ovarian failure (POF) induced by cyclophosphamide. Methods 6 SD rats were used as control group, and 12 SD rats were randomly divided into POF group and puerarin group after modeling, with 6 rats in each group. Using cyclophosphamide to induce a POF model in rats, the puerarin group received an intraperitoneal injection of 50 mg/kg puerarin, while the control group and POF group received an equal amount of saline via intraperitoneal injection, with ongoing intervention for 21 days. Cyclophosphamide induced POF model was established in rats, 50 mg/kg puerarin was injected intraperitoneally, and the changes of estrous cycle were observed by vaginal smear. Detection of serum anti-Mullerian hormone (AMH), Estradiol (E2), Luteinizing hormone (Luteinizing hormone) by enzyme-linked immunosorbent assay LH) and Follicle-stimulating hormone (FSH) levels, hematoxylin-eosin staining were used to observe the pathological changes of ovarian tissue, and the number of autophagosomes in ovarian tissue was detected by transmission electron microscopy. The expression level of microtubule associated protein light chain 3-Ⅱ (LC3-Ⅱ) was determined by immunohistochemistry. The expressions of Beclin 1 (Beclin 1) and autophagy adapter protein 62 (p62) were detected by Western blot. Results Compared with the control group, after cyclophosphamide induction, the estrous cycle of rats in POF group was disturbed, the volume of ovarian tissue was atrophied, and the number of follicles was reduced. The contents of AMH [(491.50±47.77) pg/mL compared with (1228.63±100.66) pg/mL, P<0.01] and E2 [(32.63±2.01) pg/mL compared with (54.65±7.37) pg/mL, P<0.01] were significantly decreased. The contents of LH [(18.43±1.16) mIU/mL compared with (10.08±0.27) mIU/mL, P<0.01] and FSH [(12.50±1.41) IU/L compared with (7.55±0.86) IU/L, P<0.01] were significantly increased. Compared with POF group, the estrous cycle of puerarin group was normal, the ovarian structure was restored, and the number of follicles was increased. The content of AMH [(859.57±159.89) pg/mL compared with (491.50±47.77) pg/mL, P<0.01] and E2 [(48.86±6.60) pg/mL compared with (32.63±2.01) pg/mL, P<0.01] were significantly decreased. The contents of LH [(12.43±1.70) mIU/mL compared with (18.43±1.16) mIU/mL, P<0.01] and FSH [(7.48±0.56) IU/L compared with (12.50±1.41) IU/L, P<0.01] were significantly increased. In addition, compared with POF group, puerarin could promote autophagosome formation and LC3-Ⅱ expression [(0.19±0.03) vs. (0.11±0.03), P<0.05]. Up-regulated Beclin 1 [(0.919±0.044) ratio (0.395±0.048), P<0.01] and down-regulated p62/GAPDH [(0.851±0.063) ratio (1.629±0.105), P<0.01] protein expression. Conclusion Puerarin can alleviate the damage of premature ovarian failure induced by cyclophosphamide by promoting autophagy. |
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