槲皮素改善多囊卵巢综合征模型大鼠胰岛素抵抗和卵泡发育的作用机制研究  

Mechanism of quercetin in improving insulin resistance and follicular development in a polycystic ovary syndrome rat model

作　　者：江雪娟 陈晓菲 俞佳[1] 王晨晔[1] 胡慧敏[1] 丁彩飞[1] JIANG Xuejuan;CHEN Xiaofei;YU Jia;WANG Chenye;HU Huimin;DING Caifei(Reproductive Medicine Department,Zhejiang Hospital of Integrated Traditional Chinese and Western Medicine,Hangzhou,Zhejiang 310003,China)

机构地区：[1]浙江省中西医结合医院生殖科,杭州310003

出　　处：《浙江中西医结合杂志》2024年第10期888-896,共9页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：杭州市医药卫生科技计划项目(No.ZD20200016、ZD20210029);浙江省中医药科技计划项目(No.2021ZB210);杭州市红十字会医院院内科研基金项目(No.HHQN2021005);浙江省自然科学基金项目(No.LQ19H270005)。

摘　　要：目的探究槲皮素是否通过高级糖基化终末产物-受体(AGEs/RAGE)代谢通路改善多囊卵巢综合征(PCOS)模型大鼠胰岛素抵抗(IR)和卵泡发育。方法30只雌性SD大鼠随机平均分为空白对照组(10%乙醇0.5 mL灌胃)、模型组(10%乙醇0.5 mL灌胃)、槲皮素低剂量组(槲皮素15 mg/kg,溶于0.5 mL 10%乙醇灌胃)、槲皮素高剂量组(槲皮素30 mg/kg,溶于0.5 mL10%乙醇灌胃)和阳性对照组(二甲双胍盐酸盐50 mg/kg,溶于0.5 mL10%乙醇灌胃)。来曲唑联合高脂饲料建立PCOS大鼠模型,给予相应药物干预4周。测定各组大鼠空腹血糖(FPG)及空腹胰岛素(FINS)并计算胰岛素抵抗指数(HOMA-IR),通过大鼠阴道涂片观察动情周期变化,酶联免疫吸附试验检测大鼠血清雌二醇(E2)、孕酮(P)、促卵泡生成素(FSH)、黄体生成素(LH)、睾酮(T)水平,苏木精-伊红(HE)染色观察大鼠卵巢组织病理变化,TUNEL法检测大鼠卵巢组织细胞凋亡情况,免疫组化及蛋白质免疫印迹(Western blot)测定AGEs/RAGE通路及凋亡相关蛋白高级糖基化终末产物(AGEs)及其受体(RAGE)、胱天蛋白酶3(Caspase-3)、裂解型胱天蛋白酶3(cleaved-Caspase-3)、B细胞淋巴瘤-2相关X蛋白(Bax)、B细胞淋巴瘤-2蛋白(Bcl-2)表达情况。结果与空白对照组比较,模型组大鼠FPG[(7.27±0.67)mmol/L比(4.57±0.34)mmol/L,P<0.01]、FINS[(23.17±1.13)mU/L比(8.88±0.26)mU/L,P<0.01]、HOMA-IR[(7.50±0.91)比(1.80±0.14),P<0.01]、血清E2[(46.02±4.07)pg/mL比(35.09±3.42)pg/mL,P<0.01]、LH[(31.08±2.76)mIU/mL比(22.17±2.06)mIU/mL,P<0.01]、T[(14.92±1.12)ng/mL比(8.26±0.73)ng/mL,P<0.01]、卵巢组织Caspase-3[(0.28±0.02)比(0.17±0.02),P<0.01]、Bax[(0.36±0.03)比(0.19±0.03),P<0.01]蛋白均显著升高,血清P[(64.25±4.91)ng/mL比(84.58±7.64)ng/mL,P<0.01]和卵巢组织Bcl-2[(0.19±0.02)比(0.25±0.02),P<0.01]表达水平显著降低,证明造模成功。与模型组比较,干预后槲皮素高剂量组大鼠FPG[(5.83±0.60)mmol/L比(7.27±0.67)mmol/L,P<0.01]、FIObjective This study aimed to investigate whether quercetin(Q)can improve insulin resistance(IR)and promote follicular development in a polycystic ovarian syndrome(PCOS)rat model through modulation of the advanced glycation end-product/receptor for advanced glycation end-product(AGEs/RAGE)metabolic pathway.Methods Thirty female SD rats were randomly and evenly divided into the blank control group(gavaged with 0.5 mL of 10%ethanol),the model group(gavaged with 0.5 mL of 10%ethanol),the low-dose quercetin group(gavaged with 15 mg/kg dissolved in 0.5 mL of 10%ethanol),the high-dose quercetin group(gavaged with 30 mg/kg,dissolved in 0.5 mL of 10%ethanol),and the positive control group(gavaged with 50 mg/kg metformin hydrochloride,dissolved in 0.5 mL of 10%ethanol).A PCOS rat model was established by letrozole combined with a high-fat diet,and intervention treatments were administered for four weeks.Fasting plasma glucose(FPG)and fasting insulin(FINS)levels were measured and the homeostasis model assessment for insulin resistance(HOMA-IR)was calculated.Estrous cycle changes were monitored through vaginal smears of the rats.The levels of estradiol(E2),progesterone prog(P),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and androgen(T)were detected using enzyme-linked immunosorbent assay.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in ovarian tissue of the rats,and terminal deoxynucleotidyl transferase dUTP nick-end labeling was used to detect apoptosis in ovarian tissue cells.The expression levels of AGEs/RAGE pathway and apoptosis-related proteins,including AGEs,RAGE,cleaved-caspase3,Bax,and Bcl-2 were determined using immunohistochemistry and western blotting.Results Compared with the blank control group,the model group showed significantly higher levels of FPG[(7.27±0.67)mmol/L vs.(4.57±0.34)mmol/L,P<0.01],FINS[(23.17±1.13)mU/L vs.(8.88±0.26)mU/L,P<0.01],and HOMA-IR[(7.50±0.91)vs.(1.80±0.14),P<0.01].Serum levels of E2[(46.02±4.07)pg/mL vs.(35.09±3.42)pg/mL,P<0.01],LH[(

关 键 词：大鼠 多囊卵巢综合征 槲皮素 卵泡 高级糖基化终末产物-受体 

分 类 号：R285.5[医药卫生—中药学]