【作者】 方云芸；

【导师】 傅春华；

【作者基本信息】 成都中医药大学 ， 中医妇科学， 2010， 硕士

【摘要】 目的：探讨通脉大生片对肾虚排卵障碍型不孕大鼠促排卵的作用机制,以阐明中医补肾方药治疗肾虚排卵障碍型不孕症的有效性。方法：建立肾虚排卵障碍型不孕大鼠模型,随机分七组,为模型组、通脉大生片高剂量组、通脉大生片中剂量组、通脉大生片低剂量组、氯米芬组、右归丸组、正常组。从第70日龄,大鼠阴道开门,连续阴道涂片两个性周期(一个性周期为4-5天),若阴道上皮无周期性变化,呈持续性角化,提示造模成功,余剔除。于大鼠第80日龄起,连续灌胃5周后股动脉取血并处死动物,迅速剥离大鼠卵巢、子宫组织。血液离心后取上清液,放入-20℃冰箱内保存,以备后期检测。大鼠阴道脱落细胞检测采用巴氏染色法,子宫、卵巢甲醛固定后、脱水包埋,HE染色,光镜下观察子宫、卵巢组织形态学,应用免疫组织化学技术SP法检测大鼠卵巢局部IGF-1R的表达。应用ELISA法检测大鼠血清内FSH、LH、IGF-1含量,应用RI法检测E2、T的含量,所有数据的统计分析均在SPSS13.0软件的支持下完成。结果：通脉大生片组与模型组比较,大鼠出现较规律的动情周期；子宫、卵巢组织形态学表明,通脉大生片组大鼠子宫内膜比模型组增厚,腺体数目增多(P<0.01),子宫内膜由增生期向分泌期转化；通脉大生片组卵巢内病理性囊性卵泡比模型组明显减少,有少量成熟卵泡和黄体生成；通脉大生片组大鼠血清中FSH、LH水平高于模型组(P<0.05),通脉大生片组血清中T水平低于模型组(P<0.05)；与模型组比较,通脉大生片具有升高血清中E2水平趋势,但差异无显著性(P>0.05)；通脉大生片组血清中IGF-1水平和卵巢局部IGF-1R的表达低于模型组(P<0.05),差异具有统计学意义。结论：1.肌注丙酸睾丸酮的雌性大鼠生殖功能下降,表现为阴道脱落细胞持续性角化,子宫内膜呈增生期,出现病理性囊性卵泡,无成熟卵泡和黄体生成,造模后血清中E2、FSH、LH水平低下,T升高,血清中IGF-1水平异常升高和卵巢局部IGF-1R表达异常增强。2.通脉大生片可以改善肾虚排卵障碍型不孕大鼠的子宫和卵巢病理形态学和纠正体内性激素水平紊乱。通脉大生片可以增加子宫内膜腺体数,增加子宫内膜厚度,促进子宫内膜从增生期转化为分泌期；通脉大生片可以降低病理性囊性卵泡数,,有成熟卵泡生成和少量黄体生成,促进卵泡的发育成熟及排出；通脉大生片能够提高血清中FSH、LH水平,具有提高血清中E2水平的趋势,降低血清中T的含量,改善垂体和卵巢的内分泌功能。3.通脉大生片对肾虚排卵障碍型不孕大鼠的卵巢局部因子IGF-1/IGF-1R的调控有一定影响。通脉大生片可以降低血清中过高的IGF-1水平,降低卵巢局部表达过高的IGF-1R。4.提示补肾中药对肾虚排卵障碍型不孕症的治疗作用,可以通过调控性激素和卵巢局部因子来实现。更多还原

【Abstract】 Objective:To study the Tongmai Dasheng pill action mechanism on renal deficiency induced anovulation infertility disorder rats, and to clarify applying Chinese herbs to treat the renal deficiency induced anovulation infertility is validity.Methods:Renal deficiency induced anovulation infertility rats were randomly divide d into seven groups, the Tongmai Dasheng pill high-dose group, Tongmai Dasheng pill medium-dose group, Tongmai Dasheng pill low-dose group, chlorine clomifene group Yougui pill group, model group, control group. At the first 70 days, rat vagina open, consecutive vaginal smears of two cycles (4-5 days a cycle), if the vaginal epith elium was sustained keratoses without cyclical changes, suggesting that the model was successful, the other rats were removed.At the 80 days, after 5 weeks continuous fed, the animals were killed,and took blood quickly from femoral artery, stripped rat ovary, uterus. Supernatant after centrifugation of blood, placed in -20℃. Refrigerator, keep it for later testing. Rat vaginal exfoliated cells were detected by Papanicolaou st ain; After formaldehyde fixation, dehydration embedding, HE staining, look at the ut erine, ovarian morphology under the light microscopy.Apply SP immunohistochemi cal technique way to detecte rat ovary IGF-1R expression. Apply ELISA assay way to detect serum FSH/LH/IGF-1 level, RI way to detecte serum E2/T content, statistical analysis of all data are in complete support of SPSS 13.0 software.Results:Compared with Tongmai Dasheng pill group and model group, rats had a reg lar estrous cycle, uterus/ovary morphology showed that Tongmai Dasheng pill group uterus endometrial became thicker, the number of glands is more than the model grou p(P<0.01), endometrium transformation from proliferative to secretory phase; Com pared with model group, pathologic cystic ovarian follicles decreased, small amouts mature follicles and corpus luteum formation. Tongmai Dasheng pill can increase serum FSH, LH level (P<0.05), has a trend to elevate serum E2 level,but the difference was not significantly(P>0.05), and can decrease serum T level (P<0.05, Tongmai Dasheng pill aslo can reduce the serum level of IGF-1 and IGF-1R expression in the ovary (P<0.05)Conclusion:1. Intramuscular injected testosterone female rats ovulatory infertility reproductive function decreased, showing persistent vaginal exfoliated cells keratosis, endometrium presents proliferative phase, ovary presents pathologic cystic follicles, no mature follicle and corpus luteum generated. After modeling, the serum E2, FSH, LH levels become lower, T increased. Serum IGF-1 levels abnormal increased, ovarian local IGF-1R also expression in abnormal increase.2. Tongmai Dasheng pill can improve renal deficiency induced anovulation infertility disorder rats uterine and ovarian pathology and retrieve the body sex hormones levels. Tongmai Dasheng pill can increase the number of endometrial glands, increase endometrial thickness, and promote from the proliferative endometrium into a secretory phase; Tongmai Dasheng pill can reduce the pathologic cystic follicle number, small amouts mature follicles and corpus luteum formation, can promote follicular development,maturation and discharge; Tongmai Dasheng pill can increase serum FSH, LH level, has the trend of increase serum E2 level, can decrease serum T level, can improve the pituitary and ovarian endocrine function.3..Tongmai Dasheng pill had a impaction to regulate on renal deficiency induced anovulation infertility disorder rats ovary local IGF-1/IGF-1R factor. Tongmai Dasheng pill also can reduce serum high IGF-1 level, decrease ovarian local high IGF-1R expression.4. Traditional Chinese Herbs treatment action on renal deficiency induced anovulation infertility disorder rats, can achieve from regulating local ovarian factors andhormone.更多还原