Zhang Cong-Hui, Wei Fang-Jiao, Wang Xiu-Wen, Wang Kun, Li Meng, Wang En-Li, Yao Jing-Chun
School of Pharmacy, Shandong University of Traditional Chinese Medicine Ji'nan 250355, China.
School of Medicine, Ocean University of China Qingdao 266100, China.
Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(20):5516-5527. doi: 10.19540/j.cnki.cjcmm.20240702.401.
This study aims to explore the therapeutic effect of Yuzhi Zhixue Granules on polycystic ovary syndrome(PCOS) in rats and explain the underlying mechanism by metabolomics. Rats were randomized into normal, model, low-, medium-, and high-dose(0.5, 1.5, and 4.5 g·kg(-1)·d(-1)) Yuzhi Zhixue Granules, and positive control(metformin, 0.2 g·kg(-1)·d(-1)) groups. The rats in other groups except the normal group were administrated with 1 mg·kg(-1)·d(-1) letrozole and fed with a high-sugar and high-fat diet for the modeling of PCOS. After 40 days of modeling, the normal and model groups received distilled water and letrozole+distilled water, respectively, and other groups received letrozole and corresponding drugs, once a day for 50 days. The oral glucose tolerance test(OGTT) was carried out and enzyme-linked immunosorbent assay(ELISA) was conducted to detect insulin release, and the radioimmunoassay was employed to measure the serum levels of follicle-stimulating hormone(FSH), luteinizing hormone(LH), estradiol(E_2), and testosterone(T). The serum levels of high density lipoprotein-cholesterol(HDL-C), low density lipoprotein-cholesterol(LDL-C), and triglyceride(TG) were determined by an automatic biochemical analyzer. The pathological changes in the ovary were observed by hematoxylin-eosin(HE) staining. The protein levels of phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), and their phosphorylated forms in the ovary were determined by Western blot. Ultra-high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS) was employed to study the fecal and serum metabolites in the rat model of PCOS. The results showed that compared with the model group, drug administration repaired the impaired glucose tolerance, enhanced the insulin sensitivity, elevated the serum levels of HDL-C and E_2, lowered the serum levels of TG and T, ameliorated the pathological changes in the ovarian tissue, and up-regulated the protein levels of p-PI3K and p-Akt in the ovarian tissue. A total of 46 differential metabolites and 10 metabolic pathways in the fecal samples were screened out, which were mainly related to the biosynthesis of unsaturated fatty acids and tyrosine metabolism. In terms of the serum metabolism, 34 differential metabolites and 15 metabolic pathways were screened out, mainly related to the biosynthesis of unsaturated fatty acids and aminoacyl-tRNA. In conclusion, Yuzhi Zhixue Granules can alleviate the disorders of glucose, lipids, and sex hormones and improve the ovarian status in the rat model of PCOS by regulating the serum and fecal metabolism and activating the PI3K/Akt pathway.
本研究旨在探讨玉芷止血颗粒对大鼠多囊卵巢综合征（PCOS）的治疗作用，并通过代谢组学解释其潜在机制。将大鼠随机分为正常组、模型组、低、中、高剂量（0.5、1.5和4.5 g·kg⁻¹·d⁻¹）玉芷止血颗粒组及阳性对照组（二甲双胍，0.2 g·kg⁻¹·d⁻¹）。除正常组外，其他组大鼠每日给予1 mg·kg⁻¹·d⁻¹来曲唑并喂以高糖高脂饮食以建立PCOS模型。造模40天后，正常组和模型组分别给予蒸馏水和来曲唑+蒸馏水，其他组给予来曲唑及相应药物，每日1次，共50天。进行口服葡萄糖耐量试验（OGTT），采用酶联免疫吸附测定（ELISA）检测胰岛素释放，并采用放射免疫测定法测定血清卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E₂）和睾酮（T）水平。采用自动生化分析仪测定血清高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）和甘油三酯（TG）水平。通过苏木精-伊红（HE）染色观察卵巢的病理变化。采用蛋白质免疫印迹法测定卵巢中磷脂酰肌醇3激酶（PI3K）、蛋白激酶B（Akt）及其磷酸化形式的蛋白水平。采用超高效液相色谱-串联质谱（LC-MS/MS）研究PCOS大鼠模型的粪便和血清代谢物。结果显示，与模型组相比，给药可修复受损的葡萄糖耐量，增强胰岛素敏感性，提高血清HDL-C和E₂水平，降低血清TG和T水平，改善卵巢组织的病理变化，并上调卵巢组织中p-PI3K和p-Akt的蛋白水平。在粪便样本中筛选出46种差异代谢物和10条代谢途径，主要与不饱和脂肪酸的生物合成和酪氨酸代谢有关。在血清代谢方面，筛选出34种差异代谢物和15条代谢途径，主要与不饱和脂肪酸的生物合成和氨酰-tRNA有关。综上所述，玉芷止血颗粒可通过调节血清和粪便代谢并激活PI3K/Akt途径，缓解PCOS大鼠模型的糖、脂和性激素紊乱，改善卵巢状态。
