通脉大生片对肾虚排卵障碍型不孕大鼠的卵巢局部因子、激素水平的调控研究
|
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。
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.
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。
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.
引文
[1]杨菁,张燕不孕症研究概况[J],国外医学保健分册,2005,16(4)：232-235.
[2]罗丽兰,不孕与不育[M].北京：人民卫生出版社1998,202.
[3]Giudice LC. Insulin-like growth factors and ovarian folliclar development. Endocr Rev,1992,13(4):641-669.
[4]Barnes RB,Namnoum AB, Rosenfield RL, etal. The role of LH and FSH in ovarian and rogen secretion and ovarian follicular development:clinical studies in a patient with isolated FSH deficiency and malticyatic ovaries[J]. Hum Reprod, 2002,17:88-91.
[5]张丽珠.正确掌握促排卵适应症[J].中国实用妇科与产科杂志,1996,11(5)：258-259.
[6]张梅.排卵障碍性不孕中医机理及中西医结合治疗的研究进展.甘肃中医[J].2007,20(7)：70-72.
[7]胡君梅.辨证与辨病治疗女性不孕症[J].南京中医药大学学报,2001,17(1)：51-52.
[8]许丽锦,罗颂平.排卵障碍的病机探讨[J].中国中医药信息志,2003,10(3)：4-5.
[9]俞瑾.无排卵性不孕症[J].中西医结合杂志,1987,7(10)：617-618.
[10]魏美娟,俞瑾.雄激素致高胰岛素与高雄激素性无排卵大鼠模型[J].生殖医学杂志,1993,2(4)：216.
[11]夏桂成.中医妇科理论与实践二[M].北京：人民卫生出版社,2003：397.
[12]张玉珍,罗颂平.罗元恺教授论治不孕不育症学术经验介绍[J].新中医,2002,34(4)：7.
[13]秦昕.益肾温经汤对肾阳虚排卵障碍大鼠的促排卵机理研究[J].甘肃中医学院学报,27(1)：20-22.
[14]何嘉琳,方晓红,高涛等.育麟颗粒对排卵障碍大鼠子宫、卵巢ER及下丘脑弓状核超微结构影响的研究.中国中医药科技,2008,15(2)：90-92.
[15]王莉,陆利民,俞瑾等.中药天癸方对雄激素致不孕大鼠促性腺激素释放激 素的影响[J].中国中西医结合杂志,2004,24：211-214.
[16]王永宏,熊瑛.补肾化瘀方对不孕大鼠促排卵及卵巢促黄体生成素受体表达的影响[J].中华中医药杂志,2009,24(5)：656-658.
[17]尹胜,尤昭玲.寿胎丸对肾虚不孕大鼠卵泡发育及排卵的影响研究[J].湖南中医杂志,2007,23(3)：92-93.
[18]陈小野.证候动物模型诊断依据的设想与评价[J].中国医药学报,,1987,2(1)：50-56.
[19]郭振球.世界传统医学诊断医学.科学出版社,1998.
[20]孙斐,俞瑾.中药天癸方对雄激素致不孕大鼠血leptin及垂体促性腺激素的影响[J].中国中西医结合杂志,1999,19(6)：350-352.
[21]张月萍.雄激素致不孕大鼠发病机制及滋肾阴药对其促排卵的作用[J].中华内分泌代谢杂志,1994,10(2)：98-101
[22]归绥琪,俞瑾,魏美娟等.补肾中药对雄激素致不孕大鼠垂体、卵巢及肾上腺作用的实验研究[J].中国中西医结合杂志,1997,17(12)：735-737.
[23]王针织,俞超芹.补肾化瘀方对雄激素致不孕大鼠卵巢局部调控因子的影响[J].中西医结合学报.2008,6(4)：361-365.
[24]崔轶凡,王庆国.证候动物模型对中医药研究的意义及方法学探讨[J].中华中医药学刊.2009,27(12)：2525-2527.
[25]卓启墀.通脉大生片治疗无排卵功血的促排卵作用的临床观察[J]. 四川中医,1987,8：42.
[26]骆丽娟.通脉大生片对雌性肾阳虚大鼠卵巢功能影响的研究[D].成都中医药大学硕士研究生论文,2006：24-27.
[27]宋帮丽.通脉大生片对肾虚排卵障碍不孕大鼠下丘脑-垂体-卵巢轴的影响[J].成都中医药大学学报.2009,32(1)43-45.
[28]周莉,亓蓉,梁莹等.不孕症患者卵巢血流灌注与临床妊娠的研究[J].中国优生与遗传杂志.2009,17(5)：135-136.
[29]Nakagawa K,Ozawa N. A reduction in intraovarian arterial blood flow resistance after ovulation is necessary to achieve p regnancy in natural cycle[J]. J Assist Rep rod Genet.2005,22 (1):9-14.
[30]Du B,Takahashi K. Usefulness of intraovarian artery pulsatility and resistance indicesmeasurement on the day of follicle asp iration for the assessment of oocyte quality[J]. Fertil Steril.2006,85 (2):366-370.
[31]罗凌,萧美茹.补肾调冲方对排卵功能障碍性不孕症患者卵泡发育及内膜的影响[J].新中医,2008,40(4)：30-32.
[32]何嘉琳,方晓红,高涛等。育麟颗粒对排卵障碍大鼠子宫、卵巢ER及下丘脑弓状核超微结构影响的研究[J].中国中医药科技,2008,15(2):90-92.
[33]Adriaens I, Cortvrindt R, and Smitz J1 Differential FSH exposure in preantral follicle culture hasmarked effects on folliculogenesis and oo-cyte developmental competence. Human Rep roduction,2004,19:398-408.
[34]葛秦生.临床生殖内分泌学：女性与男性[M].北京：科学技术文献出版社,2001,194：226-228.
[35]Abel MH.WooLLon AN, Wilkins V, et al. The effect of a null mutation in the follicle—stimulating hormone receptor gene on mouse reproduction. Endocrinol-ogy,2000,141(5):1795-1803.
[36]梁洋,孙兴参,胡小梅等.FSH对腔前卵泡生长发育的影响.四川动物,2007,26(4)：782-785.
[37]谢宁,蔡骐浓,关惠波.补肾丹对成年雌性大鼠下丘脑—垂体—性腺轴的影响[J].黑龙江医学,2009,33(1)：48-50.
[38]BILLIG H, FURUTA I, HSUEHA JW. Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis. Endocrinology,1993,133:2204-2212.
[39]Kezele P, Skinner MK. Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone:endocrine model of follicle assembly.
[40]李诵弦,余传鑫.实用妇科内分泌学[M].上海医科大学出版社,2004：27.
[41]陈秀萍,陈锋剑,姜勋平等.鹅卵泡颗粒细胞凋亡与卵黄E2和P4水平关系
的研究[J].扬州大学学报(农业与生命科学版),2008,29(2)：30-34.
[42]魏兆莲,李美芝,陈咏健.高雄激素血症对子宫内膜功能的影响.中国妇产科临床,2002,3(3)：147.
[43]魏玉芳,王长海,刘钰林等.补肾法对大鼠绝经后骨质疏松模型的影响[J].第 四军医大学学报,2007,28(6)：544-545.
[44]董莉,姜琳,林晓倩等.补肾活血方对卵巢早衰患者血清FSH、LH、E2及PRL的影响[J].上海中医药杂志,2007,41(11)：48-49.
[45]王琳,连方,马凤梅等.二至天癸颗粒含药血清对初老小鼠卵巢颗粒细胞分泌功能的影响[J].中国中医药信息杂志,2008,15(12)：47-48.
[46]杜惠兰,宋翠淼, 马惠荣等.补肾调经方对雄激素致不孕大鼠子宫一氧化氮、血清睾酮的影响[J].中成药,2007,29(2)：284-285.
[47]孙嘉敏,王张,周力等.调经赞育丸对雄激素致不孕大鼠的影响[J].中药药理与临床,2009,25(3)：8-10.
[48]J.A. Simon, J.A. Safety of estrogen/androgen regimens, J. Reprod. Med.2001,46 (3)281-290.
[49]W.Somboonporn,S.R.Davis.National Health and Medical Research Council, Test-osterone effects on the breast:implications for testosterone therapy for women, E-ndo. Rev.2004,25:374-388.
[50]el-Roeiy A, Chen X, RobertsVJ, et al. Exp ression of insulin-like growth factor-Ⅰ(IGF-Ⅰ) and the IGF-Ⅱ and the IGF-Ⅰ, IGF-Ⅱ and insulin receptor genes and localization of the gene products in the human ovary [J]. J Clin Endocrinol Metab,1993,77:1411-1418.
[51]Pellegrini S,etal.In vivo studies on ovarian insulin-like growthfactor I concentrations in human preovulatory follicles and human ovarian circulation[J]. Hum Reprod,1995,10:1341-1345.
[52]Amato G, Izzo A, Tucker A, et al. insulin-like growth factor-binding protein-3 reduction in follicular fluid in spontaneous and stimulated cycles[J]. Fertil Steril, 1998,70:141-144.
[53]魏志新,张丽珠,李美芝等.促超排卵周期胰岛素样生长因子-Ⅰ对卵泡发育的调控研究[J].中华妇产科杂志,1997,32(8)：486-489.
[54]Tsuchiya M, Minegishi T, Kishi H, et al. Control of the expression of luteinizing hormone receptor by local factors in rat granulose cells. Arch Biochem Biophys, 1999,367 (2):185.
[55]纪巍,辛晓燕,陈必良.IGF-1及其受体在子宫平滑肌瘤中的表达以及与雌孕激素关系[J].临床军医杂志,2009,37(6)：1062-1064.
[56]Chara K,Sugimot oT.The action of GH/IGF-1/IGF-BP in osteoctasts and osteoclast.Horm Res,1997,48:45-49.
[57]王治鸿.多囊卵巢综合征高雄激素血症的形成[J].国外医学·妇产科学分册,2001,28(3)：167.
[58]丁原全,薛蓬,马乾章等.补肾复方对围绝经期大鼠卵巢胰岛素样生长因子一1 mRNA的影响[J].中国老年医学杂志,2008,28(21)：2157-2158.
[59]姜学明,凌泽喜.补肾健脾活血方对卵巢大鼠血清IGF-1的影响[J].湖北中医杂志,2005,27(11)：5-6.
[60]连方,孙振高,穆琳等.二至天癸颗粒提高卵细胞质量与小鼠卵巢内IGF-1R mRNA表达量关系的研究[J].中国中西医结合杂志,2006,26(5)：431-434.
[1]乐杰.妇产科学(第五版)[M].北京：人民卫生出版社,2001.418.
[2]L ok I H, Cheung L P, Haines C J. Ma nagement of inf ertility [J]. The Hong Kong Practitioner,2003,25:2492261.
[3]连方,杜晓果.1186例女性不孕症病因分析[J].中国优生与遗传杂志,2010,18(2)：110-112.
[4]顾春晓.李丽芸教授补肾调周法治疗排卵障碍性不孕经验[J].河南中医2004,24(1)：20-21.
[5]张晓春.归肾丸合四逆散治疗排卵障碍性不孕症59例[J].山东中医杂志,2002,21(7)：404-405.
[6]姜向坤,李云,张丽娟等.排卵功能障碍性不孕的机制探讨[J].山东中医杂志,2000,19(6)：326.
[7]陈芷玲.调经促排助孕汤治疗无排卵性不孕症32例[J].实用中医杂志,2002,18(4)：14.
[8]王志俊.255例不孕症临床观察[J],新中医,1995,27(5)：37-38.
[9]王奎武,李娜,杜平等.李广文教授治疗无排卵性不孕症54例经验[J].河南中医,1995(3)：154.
[10]刘涓.补肾活血助孕汤对卵泡发育及子宫内膜的影响[J].中国中医药信息杂志,2006,13(10)：14-16.
[11]王玉东.补肾活血促卵泡发育的临床研究[J].山东中医药大学学报,2002,26(3)：207-209.
[12]陆华,刘敏如,李春梅.养精汤促卵泡发育的临床观察[J].中国中西医结合杂志,1998,18(4)：217-220.
[13]范春茹,车胜男,崔兆琴.补肾法治疗排卵功能障碍103例[J].陕西中医,2002,23(5)：389-390.
[14]周丽蓉,俞瑾.补肾阴药治疗高胰岛素高雄激素列排卵症的临床观察[J].中国中西结合杂志,1996,16(9)：515-518.
[15]章勤,何少山.治疗女性不孕的经验[J].中医杂志,1999,40(7)：400.
[16]刘彬,李崇爱.调经助孕中药对排卵障碍性不孕者子宫内膜组织形态的影响[J].中国计划生育学杂志.2007,145(11)：684-686.
[17]林雪娟,李灿东,高碧珍等.不孕症肝郁病理与性腺轴功能变化相关性研究[J].辽宁中医杂志,2005,32(9)：874-876.
[18]黄莉萍.调肝汤治疗肝郁型不孕症41例[J].湖北中医杂志,1994,16(12)：13.
[19]李灿东,林雪娟,兰启防等.不孕症肝郁病理的生殖内分泌实质研究[J].河南中医学院学报.2004,19(115)：18-20.
[20]董纪翠.电针治疗卵泡发育不良临床观察[J].中国中医药信息杂志,2007,14(9)：68-69.
[21]马瑞芬,陆海娟,陆金霞等.针刺促排卵与血中FSH、LH、E2的关系[J].浙江中西医结合杂志,2006,(11)：711-712.
[22]俞谨,郑怀美,邴圣民.电针促排卵和血FSH、LH水平及滤泡生长的观察[J].中西医结合杂志,1998,9(4)：199-202.
[23]廖柏松,胡燕,鞠躬.二仙汤对18月龄雌性大鼠下丘脑EOP含量及其mRNA水平作用的实验研究[J].山东中医学院学报,1996,20(6)：399-401.
[24]王莉,俞瑾.中药天癸方对雄激素致不孕大鼠下丘脑前阿黑皮素mRNA的影响[J].中国中西医结合杂志,2001,21(7)：519-521.
[25]刘金星,刘敏如,宋韬等.养精汤促排卵的临床及实验研究[J].中国中西医结合杂志,2001,21(2)：94-98.
[26]何嘉琳,高涛,赵宏利.育龄颗粒对排卵障碍大鼠下丘脑神经元E R a表达的影响[J].中华中医药学刊,2007,25(9)：1881.
[27]何嘉琳,方晓红,高涛.育龄颗粒对排卵障碍大鼠大鼠子宫、卵巢E R及下丘脑弓状核超微结构影响的研究.中药研究与开发,2008,15(2)：90-92.
[28]孙裴,俞瑾.中药天癸方对雄激素致不孕大鼠血Leptin及垂体促性腺激素的影响[J].中国中西医结合杂志,1999,19(6)：350-352.
[29]归绥琪,俞瑾,魏美娟等.补肾中药对雄激素致不孕大鼠垂体、卵巢及肾上腺作用的实验研究[J].中国中西医结合杂志,1997,17(12)：735-737.
[30]王永周,王泽琛,杨敏等.天癸宝颗粒治疗排卵障碍性不孕症的药效学研究 [J].辽宁中医药大学学报,2006,8(6)：160-161.
[31]马灵芝.促排卵汤对高雄激素所致不孕小鼠的实验研究[J].河北中医,2004,26(7)：554-556.
[32]徐晓娟,金沈锐,秦旭华.右归丸水提液对小鼠卵巢颗粒细胞雌激素、孕酮分泌的影响及机制[J].四川中医,2006,24(5)：22-23.
[33]谷长任,张玮,周健秋等.养精种玉汤对兔卵巢颗粒细胞孕酮生成的影响及机制[J].中医药学报,2002,30(3)：55-56.
[34]李翠萍,黄霞,牛丽丽等.补肾调周法对PCO大鼠排卵障碍干预的实验研究[J].山西中医2008,24(5)：46-47.
[35]宋翠淼,杜惠兰,马爱蕊等.补肾调经方对雄激素致不孕大鼠子宫转化生长因子β1及其受体的影响[J].中成药,2007,29(5)：652-655.
[36]夏天,付于,韩冰等.补肾调冲方含药血清对高雄激素培养大鼠卵巢颗粒细胞增殖与分泌的影响[J].中国中西医结合杂志.2006,26(4)：348-351.
[37]王针织,俞超芹.补肾化瘀祛痰方对雄激素致不孕大鼠卵巢局部调控因子的影响[J].中西医结合学报.2008,6(4)：361-365.
[38]连方,孙振高,穆琳等.二至天癸颗粒提高卵细胞质量与小鼠卵巢内IGF-1RmRNA表达量关系的研究[J].中国中西医结合杂志.2006,26(5)：431-434.
[39]吴克明,王家葵,刘钺等.新加归肾丸对雷公藤致卵巢功能损害小鼠模型的影响[J].成都中医药大学学报.2009,32(2)：47-50.
[40]刘海燕,唐金强,袁懿等.促排卵汤对不孕大鼠卵巢局部基质金属蛋白酶mRNA表达的研究[J].湖南中医学院学报.2004,24(4)：18-19.
[41]Hu Z,Chen B, Tong J,etal. The change of c-fos expression in ovariectomized rats following elect roacupuncture treatment-an immunohistochemistry study. Acupunct Elect rother Res,1993,18 (2):117.
[42]Greisen S, Flyvbjerg A, Ledet T, et al. Regulation of insulin-like growth factor binding protein secretion by human granulosa luteal cells in a polycystic ovary-li-ke environment. Fertil Steril,2002,78:162.
[43]Petti F, Bangrazi A, Liguori A, etal. Effects of acupuncture on immune response related to opioid-like peptides. J Tradit Chin Med,1998,18 (1):55.
[44]Ulett GA,Han S,Han J S.Elect roacupuncture:mechanisms and clinical application. Biolpsychiatry,1998,44:129.
[45]Ku Y, Chang Y. Beta2endorphin and GABA2mediated depressor effect of specific elect roacupuncture surpasses pressor response of emotional circuit. Peptid-es,2001,22(9):1465.
[46]俞瑾,郑怀美,邴圣民.电针促排卵和血FSH、LH水平及滤泡生长的观察[J].中西医结合杂志,1998,9(4)：199-202.
[47]程丽娜,杜桂珍,陈伯英等.电针调整去卵巢大鼠下丘脑—垂体—卵巢轴异常功能的生化机制[J].上海针灸杂志,2001,20(6)：32-34.
[48]王兴桂,付强,李敏等电针不同穴位对排卵障碍大鼠阴道细胞学检查及雌二醇水平的影响[J].上海针灸杂志.2003,22(7)：35-36.
[49]刘金星,刘敏如,宋韬等.养精汤促排卵的临床及实验研究[J].中国中西结合杂志,2001,21(2)：94-98.
[50]沈映君.中药药理学.北京.人民卫生出版社,2000,904.
[51]沈映君.中药药理学.北京.人民卫生出版社,2000,927.
[2]罗丽兰,不孕与不育[M].北京：人民卫生出版社1998,202.
[3]Giudice LC. Insulin-like growth factors and ovarian folliclar development. Endocr Rev,1992,13(4):641-669.
[4]Barnes RB,Namnoum AB, Rosenfield RL, etal. The role of LH and FSH in ovarian and rogen secretion and ovarian follicular development:clinical studies in a patient with isolated FSH deficiency and malticyatic ovaries[J]. Hum Reprod, 2002,17:88-91.
[5]张丽珠.正确掌握促排卵适应症[J].中国实用妇科与产科杂志,1996,11(5)：258-259.
[6]张梅.排卵障碍性不孕中医机理及中西医结合治疗的研究进展.甘肃中医[J].2007,20(7)：70-72.
[7]胡君梅.辨证与辨病治疗女性不孕症[J].南京中医药大学学报,2001,17(1)：51-52.
[8]许丽锦,罗颂平.排卵障碍的病机探讨[J].中国中医药信息志,2003,10(3)：4-5.
[9]俞瑾.无排卵性不孕症[J].中西医结合杂志,1987,7(10)：617-618.
[10]魏美娟,俞瑾.雄激素致高胰岛素与高雄激素性无排卵大鼠模型[J].生殖医学杂志,1993,2(4)：216.
[11]夏桂成.中医妇科理论与实践二[M].北京：人民卫生出版社,2003：397.
[12]张玉珍,罗颂平.罗元恺教授论治不孕不育症学术经验介绍[J].新中医,2002,34(4)：7.
[13]秦昕.益肾温经汤对肾阳虚排卵障碍大鼠的促排卵机理研究[J].甘肃中医学院学报,27(1)：20-22.
[14]何嘉琳,方晓红,高涛等.育麟颗粒对排卵障碍大鼠子宫、卵巢ER及下丘脑弓状核超微结构影响的研究.中国中医药科技,2008,15(2)：90-92.
[15]王莉,陆利民,俞瑾等.中药天癸方对雄激素致不孕大鼠促性腺激素释放激 素的影响[J].中国中西医结合杂志,2004,24：211-214.
[16]王永宏,熊瑛.补肾化瘀方对不孕大鼠促排卵及卵巢促黄体生成素受体表达的影响[J].中华中医药杂志,2009,24(5)：656-658.
[17]尹胜,尤昭玲.寿胎丸对肾虚不孕大鼠卵泡发育及排卵的影响研究[J].湖南中医杂志,2007,23(3)：92-93.
[18]陈小野.证候动物模型诊断依据的设想与评价[J].中国医药学报,,1987,2(1)：50-56.
[19]郭振球.世界传统医学诊断医学.科学出版社,1998.
[20]孙斐,俞瑾.中药天癸方对雄激素致不孕大鼠血leptin及垂体促性腺激素的影响[J].中国中西医结合杂志,1999,19(6)：350-352.
[21]张月萍.雄激素致不孕大鼠发病机制及滋肾阴药对其促排卵的作用[J].中华内分泌代谢杂志,1994,10(2)：98-101
[22]归绥琪,俞瑾,魏美娟等.补肾中药对雄激素致不孕大鼠垂体、卵巢及肾上腺作用的实验研究[J].中国中西医结合杂志,1997,17(12)：735-737.
[23]王针织,俞超芹.补肾化瘀方对雄激素致不孕大鼠卵巢局部调控因子的影响[J].中西医结合学报.2008,6(4)：361-365.
[24]崔轶凡,王庆国.证候动物模型对中医药研究的意义及方法学探讨[J].中华中医药学刊.2009,27(12)：2525-2527.
[25]卓启墀.通脉大生片治疗无排卵功血的促排卵作用的临床观察[J]. 四川中医,1987,8：42.
[26]骆丽娟.通脉大生片对雌性肾阳虚大鼠卵巢功能影响的研究[D].成都中医药大学硕士研究生论文,2006：24-27.
[27]宋帮丽.通脉大生片对肾虚排卵障碍不孕大鼠下丘脑-垂体-卵巢轴的影响[J].成都中医药大学学报.2009,32(1)43-45.
[28]周莉,亓蓉,梁莹等.不孕症患者卵巢血流灌注与临床妊娠的研究[J].中国优生与遗传杂志.2009,17(5)：135-136.
[29]Nakagawa K,Ozawa N. A reduction in intraovarian arterial blood flow resistance after ovulation is necessary to achieve p regnancy in natural cycle[J]. J Assist Rep rod Genet.2005,22 (1):9-14.
[30]Du B,Takahashi K. Usefulness of intraovarian artery pulsatility and resistance indicesmeasurement on the day of follicle asp iration for the assessment of oocyte quality[J]. Fertil Steril.2006,85 (2):366-370.
[31]罗凌,萧美茹.补肾调冲方对排卵功能障碍性不孕症患者卵泡发育及内膜的影响[J].新中医,2008,40(4)：30-32.
[32]何嘉琳,方晓红,高涛等。育麟颗粒对排卵障碍大鼠子宫、卵巢ER及下丘脑弓状核超微结构影响的研究[J].中国中医药科技,2008,15(2):90-92.
[33]Adriaens I, Cortvrindt R, and Smitz J1 Differential FSH exposure in preantral follicle culture hasmarked effects on folliculogenesis and oo-cyte developmental competence. Human Rep roduction,2004,19:398-408.
[34]葛秦生.临床生殖内分泌学：女性与男性[M].北京：科学技术文献出版社,2001,194：226-228.
[35]Abel MH.WooLLon AN, Wilkins V, et al. The effect of a null mutation in the follicle—stimulating hormone receptor gene on mouse reproduction. Endocrinol-ogy,2000,141(5):1795-1803.
[36]梁洋,孙兴参,胡小梅等.FSH对腔前卵泡生长发育的影响.四川动物,2007,26(4)：782-785.
[37]谢宁,蔡骐浓,关惠波.补肾丹对成年雌性大鼠下丘脑—垂体—性腺轴的影响[J].黑龙江医学,2009,33(1)：48-50.
[38]BILLIG H, FURUTA I, HSUEHA JW. Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis. Endocrinology,1993,133:2204-2212.
[39]Kezele P, Skinner MK. Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone:endocrine model of follicle assembly.
[40]李诵弦,余传鑫.实用妇科内分泌学[M].上海医科大学出版社,2004：27.
[41]陈秀萍,陈锋剑,姜勋平等.鹅卵泡颗粒细胞凋亡与卵黄E2和P4水平关系
的研究[J].扬州大学学报(农业与生命科学版),2008,29(2)：30-34.
[42]魏兆莲,李美芝,陈咏健.高雄激素血症对子宫内膜功能的影响.中国妇产科临床,2002,3(3)：147.
[43]魏玉芳,王长海,刘钰林等.补肾法对大鼠绝经后骨质疏松模型的影响[J].第 四军医大学学报,2007,28(6)：544-545.
[44]董莉,姜琳,林晓倩等.补肾活血方对卵巢早衰患者血清FSH、LH、E2及PRL的影响[J].上海中医药杂志,2007,41(11)：48-49.
[45]王琳,连方,马凤梅等.二至天癸颗粒含药血清对初老小鼠卵巢颗粒细胞分泌功能的影响[J].中国中医药信息杂志,2008,15(12)：47-48.
[46]杜惠兰,宋翠淼, 马惠荣等.补肾调经方对雄激素致不孕大鼠子宫一氧化氮、血清睾酮的影响[J].中成药,2007,29(2)：284-285.
[47]孙嘉敏,王张,周力等.调经赞育丸对雄激素致不孕大鼠的影响[J].中药药理与临床,2009,25(3)：8-10.
[48]J.A. Simon, J.A. Safety of estrogen/androgen regimens, J. Reprod. Med.2001,46 (3)281-290.
[49]W.Somboonporn,S.R.Davis.National Health and Medical Research Council, Test-osterone effects on the breast:implications for testosterone therapy for women, E-ndo. Rev.2004,25:374-388.
[50]el-Roeiy A, Chen X, RobertsVJ, et al. Exp ression of insulin-like growth factor-Ⅰ(IGF-Ⅰ) and the IGF-Ⅱ and the IGF-Ⅰ, IGF-Ⅱ and insulin receptor genes and localization of the gene products in the human ovary [J]. J Clin Endocrinol Metab,1993,77:1411-1418.
[51]Pellegrini S,etal.In vivo studies on ovarian insulin-like growthfactor I concentrations in human preovulatory follicles and human ovarian circulation[J]. Hum Reprod,1995,10:1341-1345.
[52]Amato G, Izzo A, Tucker A, et al. insulin-like growth factor-binding protein-3 reduction in follicular fluid in spontaneous and stimulated cycles[J]. Fertil Steril, 1998,70:141-144.
[53]魏志新,张丽珠,李美芝等.促超排卵周期胰岛素样生长因子-Ⅰ对卵泡发育的调控研究[J].中华妇产科杂志,1997,32(8)：486-489.
[54]Tsuchiya M, Minegishi T, Kishi H, et al. Control of the expression of luteinizing hormone receptor by local factors in rat granulose cells. Arch Biochem Biophys, 1999,367 (2):185.
[55]纪巍,辛晓燕,陈必良.IGF-1及其受体在子宫平滑肌瘤中的表达以及与雌孕激素关系[J].临床军医杂志,2009,37(6)：1062-1064.
[56]Chara K,Sugimot oT.The action of GH/IGF-1/IGF-BP in osteoctasts and osteoclast.Horm Res,1997,48:45-49.
[57]王治鸿.多囊卵巢综合征高雄激素血症的形成[J].国外医学·妇产科学分册,2001,28(3)：167.
[58]丁原全,薛蓬,马乾章等.补肾复方对围绝经期大鼠卵巢胰岛素样生长因子一1 mRNA的影响[J].中国老年医学杂志,2008,28(21)：2157-2158.
[59]姜学明,凌泽喜.补肾健脾活血方对卵巢大鼠血清IGF-1的影响[J].湖北中医杂志,2005,27(11)：5-6.
[60]连方,孙振高,穆琳等.二至天癸颗粒提高卵细胞质量与小鼠卵巢内IGF-1R mRNA表达量关系的研究[J].中国中西医结合杂志,2006,26(5)：431-434.
[1]乐杰.妇产科学(第五版)[M].北京：人民卫生出版社,2001.418.
[2]L ok I H, Cheung L P, Haines C J. Ma nagement of inf ertility [J]. The Hong Kong Practitioner,2003,25:2492261.
[3]连方,杜晓果.1186例女性不孕症病因分析[J].中国优生与遗传杂志,2010,18(2)：110-112.
[4]顾春晓.李丽芸教授补肾调周法治疗排卵障碍性不孕经验[J].河南中医2004,24(1)：20-21.
[5]张晓春.归肾丸合四逆散治疗排卵障碍性不孕症59例[J].山东中医杂志,2002,21(7)：404-405.
[6]姜向坤,李云,张丽娟等.排卵功能障碍性不孕的机制探讨[J].山东中医杂志,2000,19(6)：326.
[7]陈芷玲.调经促排助孕汤治疗无排卵性不孕症32例[J].实用中医杂志,2002,18(4)：14.
[8]王志俊.255例不孕症临床观察[J],新中医,1995,27(5)：37-38.
[9]王奎武,李娜,杜平等.李广文教授治疗无排卵性不孕症54例经验[J].河南中医,1995(3)：154.
[10]刘涓.补肾活血助孕汤对卵泡发育及子宫内膜的影响[J].中国中医药信息杂志,2006,13(10)：14-16.
[11]王玉东.补肾活血促卵泡发育的临床研究[J].山东中医药大学学报,2002,26(3)：207-209.
[12]陆华,刘敏如,李春梅.养精汤促卵泡发育的临床观察[J].中国中西医结合杂志,1998,18(4)：217-220.
[13]范春茹,车胜男,崔兆琴.补肾法治疗排卵功能障碍103例[J].陕西中医,2002,23(5)：389-390.
[14]周丽蓉,俞瑾.补肾阴药治疗高胰岛素高雄激素列排卵症的临床观察[J].中国中西结合杂志,1996,16(9)：515-518.
[15]章勤,何少山.治疗女性不孕的经验[J].中医杂志,1999,40(7)：400.
[16]刘彬,李崇爱.调经助孕中药对排卵障碍性不孕者子宫内膜组织形态的影响[J].中国计划生育学杂志.2007,145(11)：684-686.
[17]林雪娟,李灿东,高碧珍等.不孕症肝郁病理与性腺轴功能变化相关性研究[J].辽宁中医杂志,2005,32(9)：874-876.
[18]黄莉萍.调肝汤治疗肝郁型不孕症41例[J].湖北中医杂志,1994,16(12)：13.
[19]李灿东,林雪娟,兰启防等.不孕症肝郁病理的生殖内分泌实质研究[J].河南中医学院学报.2004,19(115)：18-20.
[20]董纪翠.电针治疗卵泡发育不良临床观察[J].中国中医药信息杂志,2007,14(9)：68-69.
[21]马瑞芬,陆海娟,陆金霞等.针刺促排卵与血中FSH、LH、E2的关系[J].浙江中西医结合杂志,2006,(11)：711-712.
[22]俞谨,郑怀美,邴圣民.电针促排卵和血FSH、LH水平及滤泡生长的观察[J].中西医结合杂志,1998,9(4)：199-202.
[23]廖柏松,胡燕,鞠躬.二仙汤对18月龄雌性大鼠下丘脑EOP含量及其mRNA水平作用的实验研究[J].山东中医学院学报,1996,20(6)：399-401.
[24]王莉,俞瑾.中药天癸方对雄激素致不孕大鼠下丘脑前阿黑皮素mRNA的影响[J].中国中西医结合杂志,2001,21(7)：519-521.
[25]刘金星,刘敏如,宋韬等.养精汤促排卵的临床及实验研究[J].中国中西医结合杂志,2001,21(2)：94-98.
[26]何嘉琳,高涛,赵宏利.育龄颗粒对排卵障碍大鼠下丘脑神经元E R a表达的影响[J].中华中医药学刊,2007,25(9)：1881.
[27]何嘉琳,方晓红,高涛.育龄颗粒对排卵障碍大鼠大鼠子宫、卵巢E R及下丘脑弓状核超微结构影响的研究.中药研究与开发,2008,15(2)：90-92.
[28]孙裴,俞瑾.中药天癸方对雄激素致不孕大鼠血Leptin及垂体促性腺激素的影响[J].中国中西医结合杂志,1999,19(6)：350-352.
[29]归绥琪,俞瑾,魏美娟等.补肾中药对雄激素致不孕大鼠垂体、卵巢及肾上腺作用的实验研究[J].中国中西医结合杂志,1997,17(12)：735-737.
[30]王永周,王泽琛,杨敏等.天癸宝颗粒治疗排卵障碍性不孕症的药效学研究 [J].辽宁中医药大学学报,2006,8(6)：160-161.
[31]马灵芝.促排卵汤对高雄激素所致不孕小鼠的实验研究[J].河北中医,2004,26(7)：554-556.
[32]徐晓娟,金沈锐,秦旭华.右归丸水提液对小鼠卵巢颗粒细胞雌激素、孕酮分泌的影响及机制[J].四川中医,2006,24(5)：22-23.
[33]谷长任,张玮,周健秋等.养精种玉汤对兔卵巢颗粒细胞孕酮生成的影响及机制[J].中医药学报,2002,30(3)：55-56.
[34]李翠萍,黄霞,牛丽丽等.补肾调周法对PCO大鼠排卵障碍干预的实验研究[J].山西中医2008,24(5)：46-47.
[35]宋翠淼,杜惠兰,马爱蕊等.补肾调经方对雄激素致不孕大鼠子宫转化生长因子β1及其受体的影响[J].中成药,2007,29(5)：652-655.
[36]夏天,付于,韩冰等.补肾调冲方含药血清对高雄激素培养大鼠卵巢颗粒细胞增殖与分泌的影响[J].中国中西医结合杂志.2006,26(4)：348-351.
[37]王针织,俞超芹.补肾化瘀祛痰方对雄激素致不孕大鼠卵巢局部调控因子的影响[J].中西医结合学报.2008,6(4)：361-365.
[38]连方,孙振高,穆琳等.二至天癸颗粒提高卵细胞质量与小鼠卵巢内IGF-1RmRNA表达量关系的研究[J].中国中西医结合杂志.2006,26(5)：431-434.
[39]吴克明,王家葵,刘钺等.新加归肾丸对雷公藤致卵巢功能损害小鼠模型的影响[J].成都中医药大学学报.2009,32(2)：47-50.
[40]刘海燕,唐金强,袁懿等.促排卵汤对不孕大鼠卵巢局部基质金属蛋白酶mRNA表达的研究[J].湖南中医学院学报.2004,24(4)：18-19.
[41]Hu Z,Chen B, Tong J,etal. The change of c-fos expression in ovariectomized rats following elect roacupuncture treatment-an immunohistochemistry study. Acupunct Elect rother Res,1993,18 (2):117.
[42]Greisen S, Flyvbjerg A, Ledet T, et al. Regulation of insulin-like growth factor binding protein secretion by human granulosa luteal cells in a polycystic ovary-li-ke environment. Fertil Steril,2002,78:162.
[43]Petti F, Bangrazi A, Liguori A, etal. Effects of acupuncture on immune response related to opioid-like peptides. J Tradit Chin Med,1998,18 (1):55.
[44]Ulett GA,Han S,Han J S.Elect roacupuncture:mechanisms and clinical application. Biolpsychiatry,1998,44:129.
[45]Ku Y, Chang Y. Beta2endorphin and GABA2mediated depressor effect of specific elect roacupuncture surpasses pressor response of emotional circuit. Peptid-es,2001,22(9):1465.
[46]俞瑾,郑怀美,邴圣民.电针促排卵和血FSH、LH水平及滤泡生长的观察[J].中西医结合杂志,1998,9(4)：199-202.
[47]程丽娜,杜桂珍,陈伯英等.电针调整去卵巢大鼠下丘脑—垂体—卵巢轴异常功能的生化机制[J].上海针灸杂志,2001,20(6)：32-34.
[48]王兴桂,付强,李敏等电针不同穴位对排卵障碍大鼠阴道细胞学检查及雌二醇水平的影响[J].上海针灸杂志.2003,22(7)：35-36.
[49]刘金星,刘敏如,宋韬等.养精汤促排卵的临床及实验研究[J].中国中西结合杂志,2001,21(2)：94-98.
[50]沈映君.中药药理学.北京.人民卫生出版社,2000,904.
[51]沈映君.中药药理学.北京.人民卫生出版社,2000,927.