目的 探究白藜芦醇(Res)联合二甲双胍(Met)对肥胖型多囊卵巢综合征(PCOS)小鼠卵巢功能的影响.方法 选取3周龄雌性昆明小鼠66只,按照随机数表法分为对照组(n=13)和造模组(n=53).造模组采用脱氢表雄酮诱导辅以高脂高糖饮食建立肥胖型PCOS模型[对照组和造模组各随机抽取3只小鼠,采用苏木精-伊红(HE)染色,显微镜观察确定PCOS造模成功],造模成功的50只小鼠按照随机数表法分为PCOS组、Met组、Res低剂量+Met组、Res中剂量+Met组和Res高剂量+Met组,每组10只,分别灌胃0.2 mL蒸馏水、0.2 mL Met[300 mg/(kg·d)]溶液、0.2 mL Res[20 mg/(kg·d)]+Met[300 mg/(kg·d)]混合液、0.2 mL Res[50 mg/(kg·d)]+Met[300 mg/(kg·d)]混合液和0.2 mL Res[80 mg/(kg·d)]+Met[300 mg/(kg·d)]混合液,持续干预28 d,称量各组小鼠体重和卵巢重量;采集血液标本检测各组卵泡刺激素(FSH)、黄体生成素(LH)、睾酮(T)水平;光学显微镜下观察各组卵巢组织病理学形态.结果 PCOS组小鼠体重和卵巢重量显著高于对照组,差异均有统计学意义(P均＜0.05);经过药物干预后,各干预组小鼠体重及卵巢重量均明显低于PCOS组,差异均有统计学意义(P均＜0.05);且Res高剂量+Met组小鼠体重及卵巢重量明显低于Met组、Res低剂量+Met组、Res中剂量+Met组,差异均有统计学意义(P均＜0.05).PCOS组小鼠血清FSH水平低于对照组,血清LH、T水平高于对照组,差异均有统计学意义(P均＜0.05);经过药物干预后,各干预组血清FSH水平均高于PCOS组,Met组、Res中剂量+Met组和Res高剂量+Met组血清LH、T水平低于PCOS组,差异均有统计学意义(P均＜0.05);且Res高剂量+Met组血清FSH水平高于Met组、Res低剂量+Met组、Res中剂量+Met组,LH、T水平低于Met组、Res低剂量+Met组、Res中剂量+Met组,差异均有统计学意义(P均＜0.05).HE染色结果显示,对照组卵巢切片显示卵巢形态正常,各类型卵泡处于不同的阶段,可见颗粒细胞和卵泡膜细胞规则和完整的组织,卵母细胞和周围的透明带正常;PCOS组大体形态均为囊性、闭锁卵泡和少量黄体,颗粒细胞层数减少,排列疏松;各药物干预组卵巢病理结构均得到改善,其中Res高剂量+Met组囊性扩张卵泡明显减少,卵母细胞和周围卵泡透明带正常且完整,接近对照组卵巢形态.与对照组比较,PCOS组原始卵泡、初级卵泡、成熟卵泡和黄体数量显著减少,次级卵泡和闭锁卵泡数量显著增多,差异均有统计学意义(P均＜0.05);经过药物干预后,与PCOS组比较,各干预组原始卵泡、初级卵泡、成熟卵泡和黄体数量增多,次级卵泡和闭锁卵泡数量减少,差异均有统计学意义(P均＜0.05);且Res高剂量+Met组原始卵泡、初级卵泡、成熟卵泡和黄体数量明显多于Met组、Res低剂量+Met组、Res中剂量+Met组,次级卵泡和闭锁卵泡数量少于Met组、Res低剂量+Met组、Res中剂量+Met组,差异均有统计学意义(P均＜0.05).结论 Res高剂量联合Met较单独应用Met可以更好地降低肥胖型PCOS小鼠体重、卵巢重量,并且能够更好地改善血清LH、FSH、T水平及卵巢卵泡发育.
The impact of resveratrol combined with metformin on ovarian function in obese polycystic ovary syndrome mice
Objective To explore the effect of resveratrol(Res)combined with metformin(Met)on ovarian function in obese polycystic ovary syndrome(PCOS)mice.Methods A total of 66 3-week-old female Kunming mice were randomly divided into the control group(n=13)and the model group(n=53).The model group was induced with dehydroepiandrosterone sulfate(DHEA)supplemented with a high-fat,high-sugar diet to establish an obese PCOS model[3 mice were randomly selected from the control group and the model group,and the success of PCOS was confirmed by hematoxylin-eosin(HE)staining microscopy].The 50 mice successfully established as an obese PCOS model were further divided into the PCOS group,Met group,low-dose Res+Met group,medium-dose Res+Met group,and high-dose Res+Met group,with 10 mice in each group.The groups were administered with 0.2 mL distilled water,0.2 mL Met[300 mg/(kg·d)]solution,0.2 mL low-dose Res[20 mg/(kg·d)]+Met[300 mg/(kg·d)]mixture,0.2 mL medium-dose Res[50 mg/(kg·d)]+Met[300 mg/(kg·d)]mixture,and 0.2 mL high-dose Res[80 mg/(kg·d)]+Met[300 mg/(kg·d)]mixture by gavage for 28 days.Body weight and ovarian weight of each group were measured.Blood samples were collected to detect levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH),and testosterone(T).Ovarian tissue pathology was observed under an optical microscope.Results The body weight and ovarian weight of the PCOS group were significantly higher than those of the control group(both P＜0.05);After drug intervention,the body weight and ovarian weight of each intervention group were significantly lower than those of the PCOS group(all P＜0.05);The body weight and ovarian weight of the high-dose Res+Met group were signifiicantly lower than those of the Met group,low-dose Res+Met group,and medium-dose Res+Met group(all P＜0.05).The serum FSH level in the PCOS group was lower,and the serum LH and T levels were higher than those in the control group(all P＜0.05);After drug intervention,the serum FSH levels of each intervention group were higher than those of the PCOS group,and the serum LH and T levels of the Met group,medium-dose Res+Met group,and high-dose Res+Met group were lower than those of the PCOS group(all P＜0.05);The serum FSH level in the high-dose Res+Met group was higher,and the LH and T levels were lower than those in the Met group,low-dose Res+Met group,and medium-dose Res+Met group(all P＜0.05).HE staining showed normal ovarian morphology in the control group,with various types of follicles at different stages,regular and complete granulosa cells,and normal oocytes with intact zona pellucida;In the PCOS group,ovaries showed cystic and atretic follicles with few corpora lutea,reduced granulosa cell layers,and loose arrangement;All drug intervention groups showed improved ovarian pathological structure,with the high-dose Res+Met group showing significantly fewer cystic follicles,normal oocytes,and intact zona pellucida,closely resembling the control group.Compared with the control group,the PCOS group had significantly fewer primordial,primary,mature follicles,and corpora lutea,and significantly more secondary and atretic follicles(all P＜0.05);After drug intervention,the number of primordial,primary,mature follicles,and corpora lutea increased,and the number of secondary and atretic follicles decreased in each intervention group compared to the PCOS group(all P＜0.05);The number of primordial,primary,mature follicles,and corpora lutea in the high-dose Res+Met group was significantly higher,and the number of secondary and atretic follicles was significantly lower than those in the Met group,low-dose Res+Met group,and medium-dose Res+Met group(all P＜0.05).Conclusion High-dose Res combined with Met was more effective than Met alone in reducing body weight and ovarian weight in obese PCOS mice,and it could better improve serum LH,FSH,and T levels and ovarian follicle development.
ResveratrolMetforminObese polycystic ovary syndromeSerum hormonesFollicular development
万方数据
维普
