二至天癸方提高体外受精—胚胎移植卵细胞质量的实验与临床研究
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摘要
目的：通过实验与临床研究，观察二至天癸方在体外受精-胚胎移植过程中对卵细胞质量的影响，并对其可能的作用机理进行探讨。方法：实验研究：将80只小鼠随机分为：1组(二至天癸方配合HMG组)，2组(二至天癸方组)，3组(HMG组)，4组(生理盐水组)，对4组进行排卵试验和卵裂试验，观察二至天癸方对卵细胞质量的影响；临床研究：将33例入选的不孕患者随机分为治疗组(二至天癸方配合西药组)和对照组(西药组)，观察治疗前后中医证候改善情况，测定注射HCG日血清雌激素(E_2)水平，对比两组卵细胞和胚胎形态学评分及受精率、卵裂率和妊娠率。结果：动物实验表明二至天癸方配合HMG组与HMG组在促排卵疗效上无显著性差异(P＞0.05)，但是在卵细胞形态学评分及受精率、卵裂率中比较有显著性差异(P＜0.05)。临床研究表明治疗组可有效的改善患者肾虚症状，症状疗效与对照组比较有显著性差异(P＜0.05)；治疗组注射HCG日血清E_2水平略低于对照组，二者比较无显著性差异(P＞0.05)，提示配合二至天癸方有减少OHSS(OHSS,ovarian hyperstimulation syndrome)发生的趋势；治疗组卵细胞及胚胎形态学评分较对照组有显著性差异(P＜0.05)，进而受精率、卵裂率、妊娠率较对照组亦有显著性差异(P＜0.05)。结论：在试管婴儿技术运用的超排卵方案中，配合二至天癸方，可与西药发挥协同作用，在保证临床促排卵疗效的前提下，提高卵细胞质量，进而提高受精率、卵裂率、妊娠率。其作用机理与促卵细胞发育、改善生殖内分泌环境、调整免疫功能及提高子宫内膜容受性的中药整体调节作用有关。
Objective: To study the effect of Erzhitiangui(EZHTG) decoction in improving quality of oocytes of IVF-ET program, and explore its acting mechanism. Methods: Experimental study : Eighty rats were randomly divided into four groups, the 1st, 2nd, 3rd and 4th group were respectively treated with EZHTG decoction combined with HMG, alone EZHTG decoction, alone HMG and 0. 9% brine. Ovulation and splitting ovums were observed to study the effect of EZHTG decoction to quality of oocytes. Clinical study: Thirty-three patients of sterility were randomly divided into the tested group(EZHTG decoction combined with western medicine) and the control group (western medicine), which were observed on improvement of symptoms of TCM before and after treatment and detected serum estradiol level on the day of injecting HCG. We compared the grades of oocyte and embryo morphology, fecuudation rate, splitting ovums rate
and pregnancy rate of the two groups. Results: Experimental study indicated that the groups of EZHTG decoction combined with HMG and alone HMG have no differences in the effect of ovulation(P>0. 05), but the former was better than the later in the grades of embryo morphology, fecuudation rate, splitting ovums rate(P < 0. 05). Clinical study showed the tested group was obviously better than the control group in improving the insufficiency of kidney (P<0. 05). As compared with the control group, serum estradiol level of the tested group on the day of injecting HCG dropped appreciably, but had no differences(P > 0.05). This suggested the tested group had the tendency of decreasing the
occurrence of OHSS . As to the grades of oocyte and embryo morphology, fecuudation rate, splitting ovums rate and pregnancy rate, there were statistically significant differences between the two groups(P<0.05). Conclusion: EZHTG decoction had cooperation with western medicine on ovulation induction. This project not only assured the effect of ovulation induction but also elevated quality of oocytes, fecuudation rate, splitting ovums rate and pregnancy rate. The effective mechanism may have relation to the integral regulating function of Chinese herbal medicine which can promote the upgrowth of oocytes,adjust immune function, improve the level of reproductive endocrine, quality of oocytes and endometrial receptivity.
Objective: To study the effect of Erzhitiangui(EZHTG) decoction in improving quality of oocytes of IVF-ET program, and explore its acting mechanism. Methods: Experimental study : Eighty rats were randomly divided into four groups, the 1st, 2nd, 3rd and 4th group were respectively treated with EZHTG decoction combined with HMG, alone EZHTG decoction, alone HMG and 0. 9% brine. Ovulation and splitting ovums were observed to study the effect of EZHTG decoction to quality of oocytes. Clinical study: Thirty-three patients of sterility were randomly divided into the tested group(EZHTG decoction combined with western medicine) and the control group (western medicine), which were observed on improvement of symptoms of TCM before and after treatment and detected serum estradiol level on the day of injecting HCG. We compared the grades of oocyte and embryo morphology, fecuudation rate, splitting ovums rate
and pregnancy rate of the two groups. Results: Experimental study indicated that the groups of EZHTG decoction combined with HMG and alone HMG have no differences in the effect of ovulation(P>0. 05), but the former was better than the later in the grades of embryo morphology, fecuudation rate, splitting ovums rate(P < 0. 05). Clinical study showed the tested group was obviously better than the control group in improving the insufficiency of kidney (P<0. 05). As compared with the control group, serum estradiol level of the tested group on the day of injecting HCG dropped appreciably, but had no differences(P > 0.05). This suggested the tested group had the tendency of decreasing the
occurrence of OHSS . As to the grades of oocyte and embryo morphology, fecuudation rate, splitting ovums rate and pregnancy rate, there were statistically significant differences between the two groups(P<0.05). Conclusion: EZHTG decoction had cooperation with western medicine on ovulation induction. This project not only assured the effect of ovulation induction but also elevated quality of oocytes, fecuudation rate, splitting ovums rate and pregnancy rate. The effective mechanism may have relation to the integral regulating function of Chinese herbal medicine which can promote the upgrowth of oocytes,adjust immune function, improve the level of reproductive endocrine, quality of oocytes and endometrial receptivity.
引文
[1] 徐淑云．药理实验方法学，北京：人民卫生出版社，1991，第2版：440～442．
[2] 李桂娴，张亚滨，史小林，等．中草药促排卵汤对小鼠胚泡数量的影响．首都医学院学报，1994；15 (3)：194～195．
[3] 乐杰．妇产科学，北京：人民卫生出版社，2000，第4版：371．
[4] 罗元恺．中医妇科学，上海：上海科学技术出版社，1990，第5版：26．
[5] 国家中医管理局颁布．中医病证诊断疗效标准，南京：南京大学出版社，1994，第1版：1．
[6] 罗丽兰．不孕与不育，人民卫生出版社，1980，第1版：450～451．
[7] 孟励．胚胎评分标准．省立医院第三届助孕技术研讨会讲义 (内部资料)．2002年5月．
[8] 卢惠霖，卢光诱．人类生殖与生殖工程，郑州：河南科学技术出版社，2001，第1版：109～110．
[9] 赵海波．促排卵药物对卵子、性激素及子宫内膜的影响．国外医学·妇产科学分册，1995，22 (3)：144．
[10] Janson KAM, Turker KE. Follicle stimulation for IVF: should GnRH agonists be our first choice?Toward Reproductive Centainty, Ferti]ity&genetics beyond 1999. The Parthenon Publishing Group, New York and London, 1999; 27: 186～194.
[11] Simon C, Gacia VJ, Valbuena D, et al. Increasing uterine receptivity by decreasing estradiol levels diring the implantaion period in high responders with the use of a follicle-stimulating hormone step down regimen. Fertil Steril, 1998; 70: 234～239.
[12] Hsu MI, KolmP, Leete J, et al. Analysis of implantation period in assisted reproduction through the use of serial human chorionic gonadotropin measurements. J Assist Pepros Cenet, 1998; 15: 496～503.
[13] Brestow RE, Karlan BY. Ovulation induction, infertility, and ovarian cancer risk. Fertil Steril, 1996; 66: 499～507.
[14] 连方，张宁，张建伟，等．中药配合体外受精及胚胎移植治疗不孕症2例．中国中西医结合杂志，2002；26(3)：627～628．
[15] 连方，张建伟，张宁，等．中西医结合疗法在试管婴儿技术中的应用．山东中医药大学学报，2002；26(3)：182～183．
[16] 连方，张宁，张建伟，等．贞芪转阴汤配合宫腔内人工授精治疗女性抗精子抗体阳性脾肾两虚型不育症的临床观察．中国中西医结合杂志，2002；22(2)：95～97．
[17] 王一丁．用上游精子宫腔内授精治疗不孕症．中国医刊，2000；35(1)：32．
[18] 孙梅，冯云，陈子江．取卵数目与卵子成熟度对IVF-ET的影响．生殖与避孕，2000；20 (3)：184～186．
[19] 李桂娴，张亚滨，许晴，等．中草药促排卵对小鼠血清Ca~(2+)等含量的影响．生殖与避孕，1996；16 (5)：383～385．
[20] 李桂娴，史小林，张亚滨，等．中草药促排卵汤对小鼠卵巢及子宫内膜作用的组织化学研究．生殖与避孕，1995；15 (6)：429～433．
[21] 张树成，沈明秀，蔡连香，等．补肾调经和养血补肾方药调经、促排卵作用的实验研究．中医杂志，1999；40(6)：369～371．
[22] 李逸平，左嘉客．哺乳类早期鼠胚发育阻滞的形成与突破．细胞生物学杂志，1992；14 (1)：135～138．
[23] 杨桂会，王佩娟，贾晓斌，等．补肾活血汤对小鼠早期胚胎发育的作用．江苏中医，2000；21 (11)：50．
[24] 刘金星，刘敏如，宋韬，等．养精汤促排卵的临床及实验研究．中国中西医结合杂志，2001；21 (2)：94．
[25] 陆华，刘敏如，李春梅．养精汤促卵细胞发育的临床观察．中国中西医结合杂志，1998；18 (4)：217．
[26] 魏美娟，俞瑾．补肾药对雄激素致无排卵大鼠垂体及卵巢的形态学变化．中国中西医结合杂志，1993；13 (3)：164．
[27] 俞瑾，孙月丽，邵公权，等．补肾化痰治疗多囊卵巢综合征中对下丘脑-垂体-卵巢功能的调节．中国中西医结合杂志，1986；6 (4)：218．
[28] 李炳如，佘运初．补肾药对下丘脑-垂体-性腺轴功能的影响．中医杂志，1984；
25 (7)：63．
[29] 田秀珠，张丽珠，杨池荪，等．血清抗精子抗体对体外受精-胚胎移植结局的影响．中华妇产科杂志，1998；33 (6)：366～367．
[30] 林建华，严隽鸿，林其德，等．自身免疫性卵巢功能衰退小鼠体液免疫和细胞免疫的测定．中华妇产科杂志，1998；33 (12)：735～736．
[31] 田秀珠，张丽珠，杨池荪，等．抗心磷脂抗体对体外受精-胚胎移植的影响．中华妇产科杂志，1998；33(2)：86～88．
[32] 赖安妮．免疫性不孕症的实验研究．中国中西医结合杂志，2000；20 (7)：491．
[33] 罗颂平．中医临床家·罗元恺．北京：中国中医药出版社，2001，第1版：86．
[34] 肖东红，杨守范．育阴灵对小鼠子宫雌孕激素受体的影响．辽宁中医杂志，1997；24 (4)：186．
[35] 钱祖淇，陆惠娟，吴苹，等．补肾育宫冲剂治疗子宫发育不良的机理研究．中国中西医结合杂志，1998；18 (4)：321．
[36] 任长征，王世华，刘建伟，等．子宫内膜雌激素受体与血清性腺轴激素含量测定对肾虚不孕症中药治疗机理的研究．山东中医学院学报，1993；17 (3)：182．
[2] 李桂娴，张亚滨，史小林，等．中草药促排卵汤对小鼠胚泡数量的影响．首都医学院学报，1994；15 (3)：194～195．
[3] 乐杰．妇产科学，北京：人民卫生出版社，2000，第4版：371．
[4] 罗元恺．中医妇科学，上海：上海科学技术出版社，1990，第5版：26．
[5] 国家中医管理局颁布．中医病证诊断疗效标准，南京：南京大学出版社，1994，第1版：1．
[6] 罗丽兰．不孕与不育，人民卫生出版社，1980，第1版：450～451．
[7] 孟励．胚胎评分标准．省立医院第三届助孕技术研讨会讲义 (内部资料)．2002年5月．
[8] 卢惠霖，卢光诱．人类生殖与生殖工程，郑州：河南科学技术出版社，2001，第1版：109～110．
[9] 赵海波．促排卵药物对卵子、性激素及子宫内膜的影响．国外医学·妇产科学分册，1995，22 (3)：144．
[10] Janson KAM, Turker KE. Follicle stimulation for IVF: should GnRH agonists be our first choice?Toward Reproductive Centainty, Ferti]ity&genetics beyond 1999. The Parthenon Publishing Group, New York and London, 1999; 27: 186～194.
[11] Simon C, Gacia VJ, Valbuena D, et al. Increasing uterine receptivity by decreasing estradiol levels diring the implantaion period in high responders with the use of a follicle-stimulating hormone step down regimen. Fertil Steril, 1998; 70: 234～239.
[12] Hsu MI, KolmP, Leete J, et al. Analysis of implantation period in assisted reproduction through the use of serial human chorionic gonadotropin measurements. J Assist Pepros Cenet, 1998; 15: 496～503.
[13] Brestow RE, Karlan BY. Ovulation induction, infertility, and ovarian cancer risk. Fertil Steril, 1996; 66: 499～507.
[14] 连方，张宁，张建伟，等．中药配合体外受精及胚胎移植治疗不孕症2例．中国中西医结合杂志，2002；26(3)：627～628．
[15] 连方，张建伟，张宁，等．中西医结合疗法在试管婴儿技术中的应用．山东中医药大学学报，2002；26(3)：182～183．
[16] 连方，张宁，张建伟，等．贞芪转阴汤配合宫腔内人工授精治疗女性抗精子抗体阳性脾肾两虚型不育症的临床观察．中国中西医结合杂志，2002；22(2)：95～97．
[17] 王一丁．用上游精子宫腔内授精治疗不孕症．中国医刊，2000；35(1)：32．
[18] 孙梅，冯云，陈子江．取卵数目与卵子成熟度对IVF-ET的影响．生殖与避孕，2000；20 (3)：184～186．
[19] 李桂娴，张亚滨，许晴，等．中草药促排卵对小鼠血清Ca~(2+)等含量的影响．生殖与避孕，1996；16 (5)：383～385．
[20] 李桂娴，史小林，张亚滨，等．中草药促排卵汤对小鼠卵巢及子宫内膜作用的组织化学研究．生殖与避孕，1995；15 (6)：429～433．
[21] 张树成，沈明秀，蔡连香，等．补肾调经和养血补肾方药调经、促排卵作用的实验研究．中医杂志，1999；40(6)：369～371．
[22] 李逸平，左嘉客．哺乳类早期鼠胚发育阻滞的形成与突破．细胞生物学杂志，1992；14 (1)：135～138．
[23] 杨桂会，王佩娟，贾晓斌，等．补肾活血汤对小鼠早期胚胎发育的作用．江苏中医，2000；21 (11)：50．
[24] 刘金星，刘敏如，宋韬，等．养精汤促排卵的临床及实验研究．中国中西医结合杂志，2001；21 (2)：94．
[25] 陆华，刘敏如，李春梅．养精汤促卵细胞发育的临床观察．中国中西医结合杂志，1998；18 (4)：217．
[26] 魏美娟，俞瑾．补肾药对雄激素致无排卵大鼠垂体及卵巢的形态学变化．中国中西医结合杂志，1993；13 (3)：164．
[27] 俞瑾，孙月丽，邵公权，等．补肾化痰治疗多囊卵巢综合征中对下丘脑-垂体-卵巢功能的调节．中国中西医结合杂志，1986；6 (4)：218．
[28] 李炳如，佘运初．补肾药对下丘脑-垂体-性腺轴功能的影响．中医杂志，1984；
25 (7)：63．
[29] 田秀珠，张丽珠，杨池荪，等．血清抗精子抗体对体外受精-胚胎移植结局的影响．中华妇产科杂志，1998；33 (6)：366～367．
[30] 林建华，严隽鸿，林其德，等．自身免疫性卵巢功能衰退小鼠体液免疫和细胞免疫的测定．中华妇产科杂志，1998；33 (12)：735～736．
[31] 田秀珠，张丽珠，杨池荪，等．抗心磷脂抗体对体外受精-胚胎移植的影响．中华妇产科杂志，1998；33(2)：86～88．
[32] 赖安妮．免疫性不孕症的实验研究．中国中西医结合杂志，2000；20 (7)：491．
[33] 罗颂平．中医临床家·罗元恺．北京：中国中医药出版社，2001，第1版：86．
[34] 肖东红，杨守范．育阴灵对小鼠子宫雌孕激素受体的影响．辽宁中医杂志，1997；24 (4)：186．
[35] 钱祖淇，陆惠娟，吴苹，等．补肾育宫冲剂治疗子宫发育不良的机理研究．中国中西医结合杂志，1998；18 (4)：321．
[36] 任长征，王世华，刘建伟，等．子宫内膜雌激素受体与血清性腺轴激素含量测定对肾虚不孕症中药治疗机理的研究．山东中医学院学报，1993；17 (3)：182．