基于微生物组学研究枸杞叶对来曲唑诱导PCOS小鼠的干预作用

The intervention effect of Lycium barbarum leaves on letrozole-induced PCOS mice based on microbiome

本研究旨在探讨枸杞叶对来曲唑诱导的多囊卵巢综合征(polycystic ovary syndrome,PCOS)小鼠的干预作用及其机制。采用来曲唑联合高脂饮食制备PCOS模型,造模成功后将40只小鼠随机分为PCOS组、阳性药二甲双胍组、枸杞叶低剂量组和枸杞叶高剂量组,灌胃给予相应药物,持续29天。实验结束摘眼球取血并收集卵巢组织,检测各组小鼠卵巢质量、空腹血糖(FBG)、空腹胰岛素(FINS)、睾酮(T)、抗苗勒管激素(AMH)、促黄体生成素(LH)、促卵泡刺激素(FSH)、雌二醇(E_(2))水平。采用苏木精-伊红染色观察卵巢组织形态,并计数卵母细胞、囊性卵泡和黄体。采集小鼠盲肠内容物进行肠道菌群组成和差异菌群等分析。动物实验过程经南京中医药大学动物伦理委员会审核通过。结果显示,PCOS小鼠动情周期紊乱。与PCOS组相比,枸杞叶组能明显减轻小鼠卵巢损伤,减少囊状扩张卵泡数,使动情周期正常化。经枸杞叶干预后的FBG、FINS、T、AMH、LH、FSH、LH/FSH水平显著降低(P<0.05),而E_(2)水平显著升高(P<0.001)。此外,枸杞叶能调节PCOS小鼠肠道菌群多样性紊乱,提高拟杆菌门丰度,降低厚壁菌门、回肠杆菌属、罗姆布茨菌属、类杆菌属的丰度。综上,枸杞叶可通过改善胰岛素抵抗、调节生殖激素紊乱及肠道菌群失调而发挥对PCOS小鼠的治疗作用,为枸杞叶的合理利用及开发提供了科学依据和有益参考。

The purpose of this study was to investigate the intervention effect and mechanism of Lycium barbarum leaves on letrozole-induced polycystic ovary syndrome(PCOS)mice.The PCOS model was prepared by letrozole combined with high-fat diet.After successful modeling,40 mice were randomly divided into PCOS group,positive drug metformin group,low-dose Lycium barbarum leaves group,and high-dose Lycium barbarum leaves group.The corresponding drugs were given by gavage for 29 days.At the end of the experiment,the eyeballs were removed for blood collection and ovarian tissue was collected.The ovarian mass,fasting blood glucose(FBG),fasting insulin(FINS),testosterone(T),anti-Mullerian hormone(AMH),luteinizing hormone(LH),follicle stimulating hormone(FSH),and estradiol(E_(2))levels were measured in each group.The morphology of ovarian tissue was observed by hematoxylin-eosin staining,and the oocytes,cystic follicles and corpus luteum were counted.Cecal contents of mice were collected for analysis of intestinal flora composition and differential flora.The animal experiment process was approved by the Animal Ethics Committee of Nanjing University of Traditional Chinese Medicine.The results showed that the estrous cycle of PCOS mice was disordered.Compared with the PCOS group,the Lycium barbarum leaves group can significantly reduce the ovarian damage of mice,reduce the number of cystic dilated follicles,and normalize the estrous cycle.After the intervention of Lycium barbarum leaves,the levels of FBG,FINS,T,AMH,LH,FSH and LH/FSH were significantly decreased(P<0.05),while the level of E_(2) was significantly increased(P<0.001).In addition,Lycium barbarum leaves can regulate the disorder of intestinal flora diversity in PCOS mice,increase the abundance of Bacteroidetes,and reduce the abundance of Firmicutes,Ileibacterium,Romboutsia and Faecalibaculum.In summary,Lycium barbarum leaves can play a therapeutic role in PCOS mice by improving insulin resistance,regulating reproductive hormone disorders and gut microbiota imbalance.It provides scientific basis and useful reference for the rational utilization and development of Lycium barbarum leaves.