PCO模型大鼠卵巢组织氧化应激水平的研究

发布时间：2018-03-11 08:21

本文选题：多囊卵巢综合征　切入点：大鼠　出处：《四川大学学报(医学版)》2015年02期 　论文类型：期刊论文

【摘要】：目的应用来曲唑建立多囊卵巢(PCO)大鼠模型,从组织及细胞水平测定模型大鼠卵巢氧化应激状态,探讨卵巢氧化应激在多囊卵巢综合征(PCOS)发病中的作用,为PCOS治疗提供新的思路。方法将6周龄清洁级雌性SD大鼠,随机编为实验组〔45只,予1%羧甲基纤维素溶液1mL/d+来曲唑1mg/(kg·d)灌胃〕和对照组(45只,仅予1%羧甲基纤维素溶液1mL/d灌胃),均持续28d。每日定时行阴道细胞涂片巴氏染色镜检,判断动情周期,每7d测量体质量了解生长情况,第29d两组大鼠统一处死、采血。测量指标:血清雌二醇(E2)、孕酮(P)、促卵泡刺激素(FSH)、促黄体生成素(LH)、睾酮(T)、性激素结合蛋白(SHBG),计算游离雄激素指数(FAI);解剖子宫、卵巢,称重后计算器官质量指数;所得两侧卵巢,一侧固定后石蜡切片HE染色,另一侧制备组织匀浆、单细胞悬液,测定组织匀浆总氧化态(TOS)、总抗氧化态(TAS)、脂质过氧化物丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力;检测卵巢单细胞悬液细胞内活性氧(ROS)水平。通过比较两组大鼠上述指标的差异,验证造模是否成功,分析卵巢组织氧化应激水平与PCOS的关系。结果 1实验组用药12~15d后动情周期消失,体质量增长明显超过对照组(P0.05);2实验组性激素改变符合人类PCOS特征;3实验组卵巢质量、卵巢指数大于对照组(P0.05),子宫质量、子宫指数小于对照组(P0.05);4相对对照组大鼠,实验组大鼠卵巢HE切片镜下表现为卵泡数量增多、白膜增厚、颗粒细胞层变薄、间质增生等改变;5实验组大鼠卵巢组织内MDA含量、TOS、氧化应激指数(OSI)高于对照组,SOD活力、TAS低于对照组(P0.05);细胞内ROS水平高于对照组(P均0.05)。结论1应用来曲唑可成功诱导大鼠PCO模型,适于研究卵巢病变。2本方法所制备的PCO模型卵巢处于明显的氧化应激状态,存在细胞氧化损伤,推测人类PCOS卵巢组织内可能也存在氧化应激,因此对于PCOS的处理,在常规药物治疗同时,应注重抗氧化治疗。
[Abstract]:Objective to study the role of ovarian oxidative stress in the pathogenesis of polycystic ovary syndrome (PCOS) in rats with polycystic ovary syndrome (PCOS) by using trazole to establish the model of PCO-induced ovarian oxidative stress in the tissue and cell levels of the model rats, and to explore the role of oxidative stress in the pathogenesis of polycystic ovary syndrome (PCOS). Methods six weeks old female SD rats of clean grade were randomly divided into experimental group (n = 45) and control group (n = 45) treated with 1% mL / d letrozole (1 mL / d) and control group (n = 45). Only 1% mL / d of carboxymethyl cellulose solution was given intragastrically for 28 days. The vaginal cell smears were examined by Pap staining every day to judge the estrous cycle, and the body mass was measured every 7 days to find out the growth of the rats. The rats in the two groups were killed at the same time on the 29th day. Measurements: serum estradiol estradiol, progesterone, follicle stimulating hormone (FSH), luteinizing hormone (LHG), testosterone (T), sex hormone binding protein (SHBG), free androgen index (FAI), anatomic uterus, ovary, organ mass index (OMI) after weighing. The tissue homogenate and single cell suspension were prepared on the other side. The total oxidative state of tissue homogenate, TASN, malondialdehyde (MDA) content, and the activity of superoxide dismutase (SOD) were measured in the tissue homogenate, the total oxidation state, the content of lipid peroxide malondialdehyde (MDA) and the activity of superoxide dismutase (SOD). The levels of reactive oxygen species (Ros) in ovarian single cell suspension cells were measured. By comparing the above indexes between the two groups of rats, it was verified whether the model was successful or not. Results 1 the oestrous cycle disappeared in the experimental group after 12 days of administration, and the body mass increased significantly than that in the control group (P 0.05). The changes of sex hormone in the experimental group were in line with the characteristics of human PCOS and the ovarian weight of the experimental group was in line with the characteristics of human PCOS. The ovarian index was higher than that of the control group (P 0.05), and the uterine weight was lower than that of the control group (P 0.05). The number of follicles, the thickness of the white membrane and the layer of granulosa cells were increased in the experimental group. The contents of MDA and oxidative stress index in ovarian tissue of experimental group were higher than that of control group (P 0.05), and the level of intracellular ROS was higher than that of control group (P 0.05). Conclusion 1 Trazole can be used to induce rat PCO model successfully. The PCO model, which is suitable for the study of ovarian pathological changes, is in a state of obvious oxidative stress and has cell oxidative damage. It is speculated that oxidative stress may also exist in the ovary of human PCOS. Therefore, the treatment of PCOS is related to the treatment of PCOS. At the same time, attention should be paid to antioxidation therapy.
【作者单位】：四川大学华西第二医院生殖内分泌科;四川大学华西医院感染性疾病中心;四川大学华西医院呼吸内科;四川大学华西第二医院检验科;四川大学华西公共卫生学院卫生统计学教研室;
【基金】：成都市科技局人口健康项目(No.12PPYD070SF-002)资助
【分类号】：R711.75;R-332

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