基于网络药理学探讨月见草油改善多囊卵巢综合征大鼠主动脉内皮损伤的机制研究  

ffect of evening primrose oil on aortic endothelial injury in rats with polycystic ovary syndrome based on network pharmacology

作　　者：刘自钰 惠亮 马文欣 刘畅 虎娜 赵帅[1] 陈冬梅[3] 杨丽 蒲静[1] 穆胜 马会明[1] LIU Ziyu;HUI Liang;MA Wenxin;LIU Chang;HU Na;ZHAO Shuai;CHEN Dongmei;YANG Li;PU Jing;MU Sheng;MA Huiming(Key Laboratory of Fertility Maintenance of Ministry of Education,Ningxia Medical University,Yinchuan 750004,China;College of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Stem Cell and Regenerative Medicine,General Hospital of Ningxia Medical University,Yinchuan 750003,China;Laboratory Animal Center,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学生育力保持教育部重点实验室,银川750004 [2]宁夏医科大学中医学院,银川750004 [3]宁夏医科大学总医院宁夏干细胞与再生医学重点实验室,银川750003 [4]宁夏医科大学实验动物中心,银川750004

出　　处：《中国实验动物学报》2025年第3期324-341,共18页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(81660813)。

摘　　要：目的 基于网络药理学和体内实验探究月见草油(evening primrose oil, EPO)对多囊卵巢综合征(polycystic ovary syndrome, PCOS)大鼠主动脉血管内皮损伤的改善作用机制。方法 通过网络药理学技术预测EPO改善PCOS大鼠主动脉内皮损伤可能的靶点,并且通过实验对筛选出的核心靶点及通路进行验证。58只雌性SD大鼠随机分为:空白对照组(10只)、造模组(48只);空白对照组给予正常饮食,造模组给予高脂饮食8周,第6周联合来曲唑(1 mg/(kg·d))灌胃21 d进行PCOS模型制备,造模结束尾静脉取血,收集血清检测激素水平后,模型大鼠随机分为4组,给予相应药物灌胃治疗6周。给药结束后收集大鼠血液、血管及卵巢组织。苏木素-伊红(HE)染色观察组织形态,ELISA检测血清中促黄体生成素(LH)、睾酮(T)、促卵泡激素(FSH)、内皮素(ET-1)、肿瘤坏死因子(TNF-α)水平,分光光度法测定大鼠血清中一氧化氮(NO)水平。Western Blot及免疫组织化学法(IHC)分别检测核心靶点与RAS通路相关因子等蛋白表达水平。结果 网络药理学分析得到EPO与PCOS交集靶点25个,KEGG结果显示EPO通过肾素-血管紧张素通路(renin-angiotensin signaling, RAS)等多条通路改善PCOS大鼠血管损伤。与模型组相比,EPO低、高剂量组血清中TNF-α、FSH、LH以及T含量均有所下降,差异具有显著性(P<0.01);EPO低、高剂量组主动脉AngⅠ、VEGF-B、AT_2R、ET-1和TNF-α蛋白表达量显著降低(P<0.01),AngⅡ、CD31蛋白显著升高,差异具有显著性(P<0.01),EPO高剂量组eNOS蛋白显著升高,差异具有显著性(P<0.01)。结论 EPO可能通过抑制RAS信号通路ACE/AngⅡ/AT_1轴过度活化改善PCOS模型大鼠血管内皮损伤。Objective To explore the effect of evening primrose oil(EPO) on aortic endothelial damage in rats with polycystic ovary syndrome(PCOS), using network pharmacology and in vivo experiments. Methods The potential targets of EPO for improving aortic endothelial injury in PCOS rats were predicted by network pharmacology, and the selected core targets and renin-angiotensin signaling(RAS) pathway were verified by experiments. Fifty-eight female SD rats were divided randomly into a blank group(n = 10) and a modeling group(n = 48). Rats in the blank group were fed a normal diet and rats in the modeling group received a high-fat diet for 8 weeks. The PCOS model was prepared at week 6 by administration of letrozole(1 mg/(kg·d)) for 21 days. Blood was taken from the tail vein after modeling and serum was collected to detect hormone levels. The model rats were then divided randomly into four groups and treated with the corresponding drugs for 6 weeks. Blood, blood vessels, and ovaries were then collected. Tissue morphology was examined by hematoxylin and eosin staining and serum levels of luteinizing hormone(LH), testosterone(T), follicle-stimulating hormone(FSH), endothelin(ET-1), and tumor necrosis factor(TNF-α) were detected by enzyme-linked immunosorbent assay(ELISA). Serum levels of nitric oxide(NO) were determined by spectrophotometry. Protein expression levels of core targets and RAS pathway-related factors were assessed by western blotting and immunohistochemistry. Results Twenty-five intersection targets of EPO and PCOS were identified by network pharmacological analysis. Kyoto encyclopedia of genes and genomes analysis showed that EPO improved vascular injury in PCOS rats via multiple pathways, including RAS. Serum levels of ET-1, FSH, LH, and T measured by ELISA were significantly decreased after EPO treatment, compared with the model group(P < 0.01). EPO significantly decreased the expression levels of AngⅠ, VEGF-B, AT_(2)R, ET-1, and TNF-α proteins in the aorta(P < 0.01) and significantly increased expressio

关 键 词：多囊卵巢综合征 肾素-血管紧张素通路 主动脉内皮损伤 月见草油 网络药理学 

分 类 号：Q95-33[生物学—动物学]