针药治疗免疫性不孕症及其对β-内啡肽调节作用的研究
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摘要
免疫性不孕约占女性不孕症患者的10%～30%,可分为同种免疫性不孕和自身免疫性不孕,同种免疫性不孕由抗精子抗体(AsAb)所致,自身免疫性不孕主要由不孕妇女血清中的抗子宫内膜抗体和透明带抗体引起。目前西医主要通过类固醇药物、避孕套疗法、人工授精法及体外授精与胚胎移植法帮助受孕,但疗效不够满意,并存在副作用大、费用高等缺点。近年来,中医药在免疫性不孕的治疗方面常获满意疗效,并且中医药副作用低,费用少,因此得到广大患者的认可。许多研究表明,中药对于免疫性不孕具有较佳临床疗效,而针灸对于人体免疫调节也有良好的作用。但将针灸与中药相结合治疗女性免疫性不孕的报道较少,同时针药结合疗法的获效机理也不十分清楚,这都成为进一步发挥中医药在免疫性不孕的治疗优势的障碍。
目的：本研究拟从神经调节的角度出发,通过临床观察和动物试验两方面的综合研究,分别采用针灸配合中药治疗(治疗组)和中药治疗(对照组)的方法进行治疗,根据受试对象治疗前后血清AsAb、β-内啡肽(β-EP)等免疫学及神经递质相关指标的变化,以及患者最后妊娠的情况,旨在观察针药治疗免疫性不孕的临床疗效及部分作用机理。
方法：
1.临床研究。按随机对照的原则,将符合本研究纳入标准的患者分为针刺组30例和中药对照组30例。两组患者均服用归肾丸(熟地黄、菟丝子各30g,女贞子、山药各15g,枸杞子、首乌、桑寄生各20g,山茱萸、淫羊藿各10g组成。加减：偏血瘀者,加三七15g,丹参30g；若偏气虚者,加党参15g,黄芪30g；若兼有生殖道炎症者,加用蒲公英30g),每日1剂,水煎,分2次服,3个月为1疗程,治疗1～2个疗程。治疗组在中药治疗的基础上给予针刺,主穴选肾俞、三阴交、太溪。配穴：血瘀者配内关、膈俞；气虚配足三里、关元；生殖道炎症配丰隆。一周治疗2次,疗程同上。
2.动物实验。将55只雌性大鼠中随机分成5组,分别为健康空白组8只,阳性空白组8只,中药组13只,针刺组13只,针药组13只。于除健康空白组以外的SD大鼠双侧腹股沟及颈背部多点皮下注射1ml抗原作为首次免疫。未产生AsAb者于首次免疫第2周用同样方法加强免疫一次。健康空白组大鼠在造模大鼠免疫同时于双侧腹股沟及颈背部多点注射生理盐水,每只每次1ml。造模成功后开始给药,连续给药4周。中药组及针药组的SD大鼠采用归肾液灌胃,剂量为每日4ml；健康空白组灌以相同容量的生理盐水。针刺组及针药组开始针刺,将大鼠固定在鼠夹,针刺肾俞、三阴交、太溪,每次留针20 min,每天1次,连续4周。
结果：临床研究中,针药组的临床疗效总有效率、ASAb的转阴率及妊娠率分别为93.33%、56.67%、36.67%,中药组组的临床疗效总有效率、ASAb的转阴率及妊娠率分别为86.67%、30%、13.33%,以及两组治疗前后血清ASAb下降水平的差异,均有统计学意义。动物实验中,针药组、中药组与阳性空白组治疗后血清β-EP水平比较,针药组与中药组治疗后血清β-EP水平比较,均有统计学意义。
结论：1.本研究中采用ELISA法测定血清AsAb水平,借助实验室检查指标诊断免疫性不孕症,用西医的诊断技术,克服中医免疫性不孕症诊断的不足。2.治疗后结合现代医学诊疗手段判断疗效,检测血清ASAb下降水平,同时观察患者妊娠情况,发现针药组治疗免疫性不孕症疗效确切,能促进患者受孕。3.针药通过调节β-EP的分泌,从而通过NIM网络调节生殖免疫功能,抑制ASAb的产生,改善患者体内生殖免疫环境。4.针药结合治疗比单纯中医或西医治疗效果佳,值得推广应用。
Immune infertility accounts for about 10% to 30% in female infertility, can be divided into Alloimmune infertility and Autoimmune infertility. Alloimmune infertility is caused by the AsAb, while Autoimmune infertility is mainly caused by anti-endometrial antibody and zona pellucida antibodies in serum of Infertility. Currently, western medicine mainly help conception through steroids, condom therapy, artificial insemination and in vitro fertilization method and embryo transplant, but the effect is not satisfied and it has the shortcomings of side effects and high cost. In recent years, traditional Chinese medicine in the treatment of immune infertility often makes satisfactory effect, and the majority of patients receive the recognition because of the low side effects and less cost. Many studies show that traditional Chinese medicine for immunological infertility have a better clinical effect, also the effect of acupuncture on the human body immune function is good.But the report which combines acupuncture and traditional Chinese medicine to treat the feminine immune infertility is very few, and the effect mechanism of combined therapy is not very clear. And for that Chinese medicine in immune infertility treatment superiority will be impeded.
Objective:This research starts from the perspective of neuromodulation. With clinical observation and animal experiments, we adopt acupuncture with herbal treatment or herbel treatment for patients. After treatment, we observe the clinical curative effect and part of the mechanism on immune infertility by acupuncture.
Method:
1. Clinical research. This study divide the patients who accord with the acceptable standards, in accordance with the principle of randomized controlled, into'Acupuncture with Herbal treatment'treatment group with 30 and'Traditional Chinese medicine'control group with30. Both two groups take traditional Chinese medicine'Guishen liquid'(Ripe rehmannia glutinosa 30g, dodder 30g,Ligustrum lucidum 15g,Chinese yam 15g, Medlar 20g,Shou Wu 20g, loranthus parasiticus 20g, dogberry 10g, herba epimedii 10g;add-subtract: partial blood stasis, add panax notoginseng 15g, Salvia miltiorrhiza 30g;if partial deficiency, add Codonopsis 15g, Astragalus membranaceus 30g;genital tract inflammation, add dandelion 30g), once a day, decoction, drink it in the morning and night, three months per course, a total of 1-2 courses. The treatment group also take acupuncture, the main points are Shenshu, Sanyinjiao, Taixi, add-subtract:blood stasis, add Geshu;if partial deficiency, add Zusanli, Guanyuan;genital tract inflammation, add Fenglong. Once every other day,30 minutes keeping every time, three months per course, a total of 1～2 courses.
2. Animal experiments.55 female rats are randomly divided into 5 groups, healthy control group8, positive control group 8, traditional Chinese medicine group 13, acupuncture group 13, acupuncture with herbal group13. Injecting lml antigen in some points in bilateral inguinal and neck back of each SD rat as the first immunization except the healthy control group. AsAb who has not been created in the first immunization should be intensified in the same way in next week. At the same time, injecting lml NS in some points in bilateral inguinal and neck back of the healthy control group, 1ml per each rat.After the modeling, we will start to give rats medicine for 4 weeks. Traditional Chinese medicine group, Acupuncture with herbal group are fed 4ml Guishen liquid daily, the healthy control group are fed 4ml NS daily. Acupuncture group, Acupuncture with Chinese medicine group start acupuncture. The rats are fixed on the rat trap, acupuncture Shenshu, Sanyinjiao, Taixi,30 minutes keeping every time,1 time per day for 4 weeks.
Results:Clinical research:In the treatment group,the total clinical effects is 93.33%, the negative rate of ASAb is 56.67%, and the pregnancy rate is 36.67%. In the control group, the total clinical effects is 86.67%, the negative rate of ASAb is 30%,and the pregnancy rate is 13.33%. and the levels of serum anti-sperm antibodies (ASAb) in the two groups after treatment has decreased. All are significant. Animal experiments:After treatment, the compares of serumβ-EP between Acupuncture with herbal group, Chinese medicine group and positive control group are significant, the compare of serumβ-EP between Acupuncture and Chinese medicine group and Chinese medicine group is also significant.
Conclusion:1. This study adopts ELISA method for detection of serum ASAb. Using laboratory diagnosis immunity infertility index, with western medicine diagnosis technology,We overcome the shortage of Chinese medicine diagnosis of immune infertility.2. After treatment, combining modern medical diagnosis, we detect the level of serum ASAb and observe whether patients are pregnant. We find that the treatment of immune infertility in treatment group is effective, it can promote pregnancy.3. Acupuncture and Chinese medicine can adjustβ-EP, thereby adjust reproductive immune function by NIM, inhibit the generation of ASAb to improve patient reproductive immune environment.4. Combined treatment of Acupuncture and Chinese medicine is better than simply Chinese medicine and Western medicine treatment. It is worth to generalize and apply as treatment method.
目的：本研究拟从神经调节的角度出发,通过临床观察和动物试验两方面的综合研究,分别采用针灸配合中药治疗(治疗组)和中药治疗(对照组)的方法进行治疗,根据受试对象治疗前后血清AsAb、β-内啡肽(β-EP)等免疫学及神经递质相关指标的变化,以及患者最后妊娠的情况,旨在观察针药治疗免疫性不孕的临床疗效及部分作用机理。
方法：
1.临床研究。按随机对照的原则,将符合本研究纳入标准的患者分为针刺组30例和中药对照组30例。两组患者均服用归肾丸(熟地黄、菟丝子各30g,女贞子、山药各15g,枸杞子、首乌、桑寄生各20g,山茱萸、淫羊藿各10g组成。加减：偏血瘀者,加三七15g,丹参30g；若偏气虚者,加党参15g,黄芪30g；若兼有生殖道炎症者,加用蒲公英30g),每日1剂,水煎,分2次服,3个月为1疗程,治疗1～2个疗程。治疗组在中药治疗的基础上给予针刺,主穴选肾俞、三阴交、太溪。配穴：血瘀者配内关、膈俞；气虚配足三里、关元；生殖道炎症配丰隆。一周治疗2次,疗程同上。
2.动物实验。将55只雌性大鼠中随机分成5组,分别为健康空白组8只,阳性空白组8只,中药组13只,针刺组13只,针药组13只。于除健康空白组以外的SD大鼠双侧腹股沟及颈背部多点皮下注射1ml抗原作为首次免疫。未产生AsAb者于首次免疫第2周用同样方法加强免疫一次。健康空白组大鼠在造模大鼠免疫同时于双侧腹股沟及颈背部多点注射生理盐水,每只每次1ml。造模成功后开始给药,连续给药4周。中药组及针药组的SD大鼠采用归肾液灌胃,剂量为每日4ml；健康空白组灌以相同容量的生理盐水。针刺组及针药组开始针刺,将大鼠固定在鼠夹,针刺肾俞、三阴交、太溪,每次留针20 min,每天1次,连续4周。
结果：临床研究中,针药组的临床疗效总有效率、ASAb的转阴率及妊娠率分别为93.33%、56.67%、36.67%,中药组组的临床疗效总有效率、ASAb的转阴率及妊娠率分别为86.67%、30%、13.33%,以及两组治疗前后血清ASAb下降水平的差异,均有统计学意义。动物实验中,针药组、中药组与阳性空白组治疗后血清β-EP水平比较,针药组与中药组治疗后血清β-EP水平比较,均有统计学意义。
结论：1.本研究中采用ELISA法测定血清AsAb水平,借助实验室检查指标诊断免疫性不孕症,用西医的诊断技术,克服中医免疫性不孕症诊断的不足。2.治疗后结合现代医学诊疗手段判断疗效,检测血清ASAb下降水平,同时观察患者妊娠情况,发现针药组治疗免疫性不孕症疗效确切,能促进患者受孕。3.针药通过调节β-EP的分泌,从而通过NIM网络调节生殖免疫功能,抑制ASAb的产生,改善患者体内生殖免疫环境。4.针药结合治疗比单纯中医或西医治疗效果佳,值得推广应用。
Immune infertility accounts for about 10% to 30% in female infertility, can be divided into Alloimmune infertility and Autoimmune infertility. Alloimmune infertility is caused by the AsAb, while Autoimmune infertility is mainly caused by anti-endometrial antibody and zona pellucida antibodies in serum of Infertility. Currently, western medicine mainly help conception through steroids, condom therapy, artificial insemination and in vitro fertilization method and embryo transplant, but the effect is not satisfied and it has the shortcomings of side effects and high cost. In recent years, traditional Chinese medicine in the treatment of immune infertility often makes satisfactory effect, and the majority of patients receive the recognition because of the low side effects and less cost. Many studies show that traditional Chinese medicine for immunological infertility have a better clinical effect, also the effect of acupuncture on the human body immune function is good.But the report which combines acupuncture and traditional Chinese medicine to treat the feminine immune infertility is very few, and the effect mechanism of combined therapy is not very clear. And for that Chinese medicine in immune infertility treatment superiority will be impeded.
Objective:This research starts from the perspective of neuromodulation. With clinical observation and animal experiments, we adopt acupuncture with herbal treatment or herbel treatment for patients. After treatment, we observe the clinical curative effect and part of the mechanism on immune infertility by acupuncture.
Method:
1. Clinical research. This study divide the patients who accord with the acceptable standards, in accordance with the principle of randomized controlled, into'Acupuncture with Herbal treatment'treatment group with 30 and'Traditional Chinese medicine'control group with30. Both two groups take traditional Chinese medicine'Guishen liquid'(Ripe rehmannia glutinosa 30g, dodder 30g,Ligustrum lucidum 15g,Chinese yam 15g, Medlar 20g,Shou Wu 20g, loranthus parasiticus 20g, dogberry 10g, herba epimedii 10g;add-subtract: partial blood stasis, add panax notoginseng 15g, Salvia miltiorrhiza 30g;if partial deficiency, add Codonopsis 15g, Astragalus membranaceus 30g;genital tract inflammation, add dandelion 30g), once a day, decoction, drink it in the morning and night, three months per course, a total of 1-2 courses. The treatment group also take acupuncture, the main points are Shenshu, Sanyinjiao, Taixi, add-subtract:blood stasis, add Geshu;if partial deficiency, add Zusanli, Guanyuan;genital tract inflammation, add Fenglong. Once every other day,30 minutes keeping every time, three months per course, a total of 1～2 courses.
2. Animal experiments.55 female rats are randomly divided into 5 groups, healthy control group8, positive control group 8, traditional Chinese medicine group 13, acupuncture group 13, acupuncture with herbal group13. Injecting lml antigen in some points in bilateral inguinal and neck back of each SD rat as the first immunization except the healthy control group. AsAb who has not been created in the first immunization should be intensified in the same way in next week. At the same time, injecting lml NS in some points in bilateral inguinal and neck back of the healthy control group, 1ml per each rat.After the modeling, we will start to give rats medicine for 4 weeks. Traditional Chinese medicine group, Acupuncture with herbal group are fed 4ml Guishen liquid daily, the healthy control group are fed 4ml NS daily. Acupuncture group, Acupuncture with Chinese medicine group start acupuncture. The rats are fixed on the rat trap, acupuncture Shenshu, Sanyinjiao, Taixi,30 minutes keeping every time,1 time per day for 4 weeks.
Results:Clinical research:In the treatment group,the total clinical effects is 93.33%, the negative rate of ASAb is 56.67%, and the pregnancy rate is 36.67%. In the control group, the total clinical effects is 86.67%, the negative rate of ASAb is 30%,and the pregnancy rate is 13.33%. and the levels of serum anti-sperm antibodies (ASAb) in the two groups after treatment has decreased. All are significant. Animal experiments:After treatment, the compares of serumβ-EP between Acupuncture with herbal group, Chinese medicine group and positive control group are significant, the compare of serumβ-EP between Acupuncture and Chinese medicine group and Chinese medicine group is also significant.
Conclusion:1. This study adopts ELISA method for detection of serum ASAb. Using laboratory diagnosis immunity infertility index, with western medicine diagnosis technology,We overcome the shortage of Chinese medicine diagnosis of immune infertility.2. After treatment, combining modern medical diagnosis, we detect the level of serum ASAb and observe whether patients are pregnant. We find that the treatment of immune infertility in treatment group is effective, it can promote pregnancy.3. Acupuncture and Chinese medicine can adjustβ-EP, thereby adjust reproductive immune function by NIM, inhibit the generation of ASAb to improve patient reproductive immune environment.4. Combined treatment of Acupuncture and Chinese medicine is better than simply Chinese medicine and Western medicine treatment. It is worth to generalize and apply as treatment method.
引文
[1]苏应宽,徐启样,江森.实用妇产科学[M],山东：山东科技出版社,1994,1：30
[2]林飞卿.医学基础免疫学[M],上海：上海医科大学出版社,1990,1：13
[3]李大金,李超荆.滋阴降火中药治疗免疫性不孕症[J].中国中西医结合杂志,1995；15(1)：3
[4]刘瑞芬,汪敏华.女性免疫性不孕症病机与治疗[J].福建中医药,2003；34(1)：26
[5]莫惠,郭慧红.免疫性不孕(AsAb阳性)中医病机探讨[J].江苏中医,1998；19(12)：8
[6]吴丽芹,王桂兰,胥京生.治疗免疫性不孕症经验[J].湖北中医杂志,2002；24(11)：14
[7]王振卿.养肝滋肾、清热解毒法治疗免疫性不孕症486例疗效观察[J].新中医,2003；35(8)：23～24
[8]李艳青,孙红,宋红湘.免疫性不孕症证治体会[J].光明中医,2007；22(10)：15
[9]田秉星,蔡秀水,李祥云.化湿消抗汤治疗免疫性不孕不育30例[J].上海中医药杂志,2002；36(2)：25-26
[10]来叶根.精子免疫性不孕症辨治探讨[J].中国中医药信息杂志,2001；8(4)：21
[11]赵凯.抗免Ⅰ号片治疗女性血清抗精子抗体阳性不孕疗效观察[J].四川中医,2001；19(4)：57～58
[12]陈文裕,姚丹霓,肖莹等.中医治疗免疫性不孕用药分析[J].新中医,2007；39(10)：74
[13]张晓峰.辨证与辨病结合治疗免疫性不孕46例临床分析[J].中国中医药杂志,2006；13(6)：60
[14]杨灵君,赵兴无,史巧英.辨证分型治疗抗精子抗体阳性56例[J].河南中医,2005；25(10)：50
[15]王春霞,李永伟.六味地黄汤加减治疗免疫性不孕症153例疗效观察[J].新中医,2008；40(2)：24
[16]张灵芳.知柏地黄汤治疗免疫性不孕54例[J].现代中西医结合杂志,2005；14(3)：293
[17]司秋荣,黄玲,宋彤云.桂枝茯苓胶囊治疗AsAb、EMAb阳性40例[J].实用中西医结合杂志,2004；4(2)：25～26
[18]马春亮,程令梅,张华.忍冬藤汤治疗抗精子抗体所致免疫性不孕47例[J].山东中医杂志,2003；22(7)：407～408
[19]郝树涛.保阴煎治疗抗精子抗体所致免疫性不孕症94例[J].新中医,2004；36(3)：55
[20]蒋莉,高仁美.补肾抑抗汤治疗免疫性不孕72例[J].四川中医,2003；21(4)：51
[21]罗宇迪,牟科媛.中西医结合治疗女性抗精子抗体阳性不孕52例[J].广西医科大学学报,2005；22(5)：783
[22]谢瑞亭,张纪云,张玮,等.中西医结合冲击治疗免疫性不孕的临床研究[J].临沂医学专科学校学报,2002；24(6)：401
[23]齐玲玲,刘春霞,胡吉英.中西医结合治疗免疫性不孕90例[J].山东中医杂志,2000；19(10)：610
[24]宋淑华.“烧山火”针刺手法治疗肾阳虚型不孕症50例[J].陕西中医,2007；28(3)：331
[25]马燕燕,杨继若.针刺促排卵治疗内分泌失调性不孕临床观察[J].中国医学理论与实践,2003；23(4)：524
[26]常静玲.针灸治疗女性不孕症32例[J].上海针灸杂志,1998；17(1)：26
[27]张继红,张慧岭,赵藏朵,等.补肾化浊法药针并用治疗免疫性不孕23例[J].陕西中医,2005；26(10)：1024～1025
[28]唐晔,贾淑华,张庆蔚.针药并用治疗抗精子抗体阳性不孕症40例[J].中国民间疗法,2001；9(9)：9-10
[29]李忠仁.实验针灸学[M],北京；中国中医药出版社,2003,2：198～200,203
[30]周雄元,王跃华.针刺调节免疫功能的机理研究进展[J].湖北中医杂志,2004；26(7)：56
[31]唐照亮,宋小鸽,侯正明,等.大鼠海马内微量注射6羟多巴胺对艾灸抗炎免疫作用的影响[J].针刺研究,2000；25(3)：184
[32]赵建础.针刺调节免疫反应途径的初步研究[J].针刺研究,1997；22(1)：91
[33]张海峡,贾成文.针灸对红细胞免疫的研究进展[J].陕西中医学院学报,2004；27(1)：22～24
[34]杨会芹,张联科.针灸对红细胞免疫功能调节的研究[J].陕西中医学院学报,2004；27(5)：76～77
[35]严伟星,郑震,王健鹤.β—内啡肽对细胞免疫功能的调节[J].南通医学院学报,1994；14(2)：134
[36]何燕萍,唐纯志,梁国珍等.针药结合疗法对免疫性不育大鼠血清抗精子抗体的影响.广州中医药大学学报,2001；18(2)：142
[37]伦新.针刺治疗对男性免疫性不育症T淋巴细胞亚群分类的影响[J].北京中医药大学学报,2004；27(4)：90
[38]何燕萍,宋阳,张小洪.归肾丸结合针刺对免疫性不育症的临床作用及对精浆酸性磷酸酶的影响[J].广州中医药大学学报,2007；24(5)：361
[39]马朱红,刘心莲,尹淑英,等.针灸对类风湿性关节炎患者免疫一神经—内分泌系统影响的临床研究[J].中国针灸,1997；16(9)：529
[40]李辉,牟晓泓,翟景慧.督脉腧穴电针对佐剂性关节炎大鼠IL-2影响的实验观察[J].针刺研究,2004；29(1)：54
[41]Saji F,Nogoro T,Ohashi K etal clinical evaluation of the enzyme-linked immunosorbent assay kitfor antisperm antibodies.Fertil steril,1988,50:644
[42]Korolkova Iv,Nikolaova MA,Boghedomov VA,etal Free radical generation in ejaculate samples from infertile patients [J].Bull Exp Biol Med,2001,13(6):555-557
[43]Myogo K,Yamano S,Nakagawa K,etal Sperm-immobilizing antibodies block capacitation in human spermatozoa [J].Arch Androl,2001,47(2):135-142
[44]王浩飞,向祖琼,王益鑫.60KD的精子膜抗原对抗精子抗体阳性精子顶体酶活性的影响[J].中华男科学杂志,2004；18(2)：15～18
[45]Shibahara H. (?)igeta M. etal.Diversity of the blocking effects of antisperm antibodies on fertilization in human and mouse.Hum Report,1996,11:2595-2599
[46]潘柏轩,龙峰.不孕不育与多种自身抗体的相关性研究[J].中华男科学杂志,2006；12(11)：1039
[47]朱长庚,刘庆莹,魏瑛,等.大鼠中枢神经元内免疫神经内分泌物质的共存[J].解剖学报,1999；30(2)：100
[48]胡电,戎霖.内源性阿片肽与子宫内膜异位不孕[J].实用妇产科杂志,1993；9(6)：308
[49]倪鑫,林葆成,王成海,等.β—内啡肽对大鼠免疫功能的影响[J].中国免疫学杂志,1993；9(1)：16
[50]Carcia I,Perez-castillo A,Moreno J,etal.β-endorphin inhibits interleukin-2 release and expression of interleukin-2 receptors in concanavalin A-stimulated splenic lymphocytes.Lymphokine Cytoline Res,1992;11:339-345
[51]王玉秀,童昭岗,胡海青.内源性阿片肽系统免疫调节功能的研究进展[J].中国临床康复,2003；7(10)：1559
[52]孙志扬,过宗南,赵孟尧.β—内啡肽对正常人红细胞免疫功能的调控实验研究[J].上海免疫学杂志,1994；14(4)：200
[53]RhanNA.FernereF,Deschaux P Serotonin—inducedcalctum via 5-HT1a receptors in Human Leukeima(R562)cells Cellul Immunol,1995.165:148
[54]LabergSCrikshankwwBeerDj.etalSecretionofIL6(lym. Phocytechemottractant factor)from serotonin—stimulated CD8+Ttells in vitro J Immunol.1996,156:310
[55]张清泉,彭聿平,邱一华,等.5—羟色胺与体液免疫应答的相互作用[J].南通医学院学报,1995；15(4)：504～505
[56]高娜,王阿敬,杨渝珍,等.阿片受体介导大鼠海马内脑啡肽对细胞免疫功能的调节[J].生理学报,1999；51(5)：106～109
[57]李晓玉.甲硫氨酸脑啡肽的免疫调节作用[J].中国药理学报,1998；19(1)：5～6
[58]严伟星,王健鹤,张清皋.纹状体中亮一脑啡肽在针刺调节免疫功能中的作用[J].生理学报,1991；43(5)：451～456
[59]张龚,陈丽萍,任芬若,等.不明原因不孕妇女宫颈粘液中抗精子抗体、IL-2和IFN-r 的测定[J].中国免疫学杂志,2000；16(2)：624
[60]余琦.女性慢性生殖道炎症与抗精子免疫的相关研究.广州中医药大学硕士学位论文,(2002届)：21
[61]金忆.不明原因不孕患者宫颈粘液白细胞介素2及肿瘤坏死因子a[J].中华妇产科杂志,1999；34(8)：499
[62]侯淑萍.不孕症患者血清TNF、IL—2、E2、FSH、LH、P、PRL水平的变化[J].淮海医药,2002；20(4)：272
[63]Tezawala BU. JohnsonP. ReesRC. Inhibition of pregnancy viability in mice following IL—2 adrninistration[J]. Immunology,1989;67:115
[64]Beckman.Js,KeoppenolWH,Nitricoxide,superoxide,andperoxynitrite:thegood,thebad,t heugly,AmJPhysiol,1996;27(cellphysio140):1424-1437
[65]Chwaliszk,Buhimschi,Garfield RE.edtal Role of nitric oxide in obstetrics,prenat Neonat Med,1996;1(3):292-329
[66]RosselliM.Nitricoxideandreproduction.HumReprod:1997,3:639-641
[67]解启发.NO在小鼠早期胚胎吸收过程中的作用[J].中国兽医学报,2002；22(1)：90～93
[68]龙振州.医学免疫学[M],北京：人民卫生出版社,1998,2：79
[69]Heas GG. Immunologic infertility:which approach are best? Cotemp Obstet Gynol.1983;22:41
[70]罗颂平,张玉珍,梁国珍,等.免疫性自然流产与免疫性不孕的临床与实验研究[J].中医杂志,1997；38(6)：351～354
[71]Bronson R. Sperm antisbodies:Their role in infertility. Fertil steril,1984,42: 171
[72]吴爱明,郭霞.清化消抗汤治疗抗精子免疫性不孕症60例[J].河南中医,2007；27(3)：50
[73]王丽,黄萍,黄晓燕,等.1020例免疫性不孕患者的治疗结果分析[J].中华妇产科杂志,1999；34(4)：234～235
[74]刘继红,章咏裳,邓荣进,等.抗精子抗体阳性不育患者外周血淋巴细胞亚群检测及其意义[J].中国男科学杂志,1990；4(4)：200
[75]赵武述等.现代临床免疫学[M],北京：人民军医出版社,1994,1：98
[76]刘静君.种子转阴颗拉治疗免疫性不孕症(抗精子抗阳性)的临床与实验研究.山东中医药大学硕士学位论文,(2002届)：29
[77]赵蓉,杨金洲,郁美娟,等.针刺对放化疗患者T细胞及其亚群影响[J].上海针灸杂志,1994；13(6)：253
[78]陈雄华,刘又香.针灸足三里、关元对阳虚大鼠免疫功能影响的比较研究[J].中 国针灸,2000；20(9)：555
[79]谭敬书,徐绍勤,李凡成.穴位敷贴治疗常年性变态反应性鼻炎的临床研究[J].湖南中医学院学报,1994；14(2)：56～57
[80]赵宁侠,高巍,黄裕新,等.电针足三里穴对免疫抑制大鼠细胞免疫影响[J].针刺研究,2002；27(1)：56.
[81]魏赞美,庄肩岚.针刺足三里治疗白细胞减少症及对免疫功能观察[J].上海针灸杂志,1996；15(6)：12
[82]邱艳明.电针对抑郁模型大鼠单胺类神经递质影响的实验研究.北京中医药大学,(2002届)：74
[83]金光亮,周东丰,苏晶.电针对慢性应激抑郁模型大鼠脑单胺类神经递质的影响[J].中华精神科杂志,1999；32(2)：220
[84]吴绪平,冯异,刘又香,等.针刺对急性脑梗塞大鼠血浆cAMP与β—EP的影响[J].中国针灸,2002；22(7)：486
[85]丁建江,张磊.头穴透刺治疗脑梗塞及对血浆中β—内啡肽含量的影响[J].时珍国医国药,2007；18(10)：2536
[86]李辉,李晓泓,张露芬,等.电针对佐剂性关节炎大鼠下丘脑CR H、IL—2、β—EP含量的影响[J].中国针灸,2005；25(11)：795
[87]杨俊,黄祖汉.针刺对大鼠β—内啡肽含量的影响[J].针灸学报,1992；16(6)：31
[88]曲丽芳,李明权,白华,等.中药复方对实验性老年高脂血症红细胞免疫粘附功能和循环免疫复合物的作用[J].中国免疫学杂志,1996；12(2)：107
[89]国家中医药管理局.中华本草,上海：上海科技出版社,1999,1：667
[90]蔡定芳,沈自尹,张玲娟,等.右归饮对皮质酮大鼠细胞免疫及细胞因子的影响[J].中国免疫学杂志,1995；11(4)：248
[91]钟雪梅,周灵,杨家林,等.补经合剂促卵泡发育和排卵的实验研究[J].江苏中医,1998；19(3)：45
[92]沈皓,蔡德培.补肾中药对下丘脑—垂体促性腺机能的影响[J].中西医结合学报,2004；1(2)：53
[93]张炜,蔡德培.补肾中药对雌性青春期大鼠垂体GnHR受体mRNA及其受体蛋白表达的影响[J].中国中西医结合杂志,2003；23(2)：120
[2]林飞卿.医学基础免疫学[M],上海：上海医科大学出版社,1990,1：13
[3]李大金,李超荆.滋阴降火中药治疗免疫性不孕症[J].中国中西医结合杂志,1995；15(1)：3
[4]刘瑞芬,汪敏华.女性免疫性不孕症病机与治疗[J].福建中医药,2003；34(1)：26
[5]莫惠,郭慧红.免疫性不孕(AsAb阳性)中医病机探讨[J].江苏中医,1998；19(12)：8
[6]吴丽芹,王桂兰,胥京生.治疗免疫性不孕症经验[J].湖北中医杂志,2002；24(11)：14
[7]王振卿.养肝滋肾、清热解毒法治疗免疫性不孕症486例疗效观察[J].新中医,2003；35(8)：23～24
[8]李艳青,孙红,宋红湘.免疫性不孕症证治体会[J].光明中医,2007；22(10)：15
[9]田秉星,蔡秀水,李祥云.化湿消抗汤治疗免疫性不孕不育30例[J].上海中医药杂志,2002；36(2)：25-26
[10]来叶根.精子免疫性不孕症辨治探讨[J].中国中医药信息杂志,2001；8(4)：21
[11]赵凯.抗免Ⅰ号片治疗女性血清抗精子抗体阳性不孕疗效观察[J].四川中医,2001；19(4)：57～58
[12]陈文裕,姚丹霓,肖莹等.中医治疗免疫性不孕用药分析[J].新中医,2007；39(10)：74
[13]张晓峰.辨证与辨病结合治疗免疫性不孕46例临床分析[J].中国中医药杂志,2006；13(6)：60
[14]杨灵君,赵兴无,史巧英.辨证分型治疗抗精子抗体阳性56例[J].河南中医,2005；25(10)：50
[15]王春霞,李永伟.六味地黄汤加减治疗免疫性不孕症153例疗效观察[J].新中医,2008；40(2)：24
[16]张灵芳.知柏地黄汤治疗免疫性不孕54例[J].现代中西医结合杂志,2005；14(3)：293
[17]司秋荣,黄玲,宋彤云.桂枝茯苓胶囊治疗AsAb、EMAb阳性40例[J].实用中西医结合杂志,2004；4(2)：25～26
[18]马春亮,程令梅,张华.忍冬藤汤治疗抗精子抗体所致免疫性不孕47例[J].山东中医杂志,2003；22(7)：407～408
[19]郝树涛.保阴煎治疗抗精子抗体所致免疫性不孕症94例[J].新中医,2004；36(3)：55
[20]蒋莉,高仁美.补肾抑抗汤治疗免疫性不孕72例[J].四川中医,2003；21(4)：51
[21]罗宇迪,牟科媛.中西医结合治疗女性抗精子抗体阳性不孕52例[J].广西医科大学学报,2005；22(5)：783
[22]谢瑞亭,张纪云,张玮,等.中西医结合冲击治疗免疫性不孕的临床研究[J].临沂医学专科学校学报,2002；24(6)：401
[23]齐玲玲,刘春霞,胡吉英.中西医结合治疗免疫性不孕90例[J].山东中医杂志,2000；19(10)：610
[24]宋淑华.“烧山火”针刺手法治疗肾阳虚型不孕症50例[J].陕西中医,2007；28(3)：331
[25]马燕燕,杨继若.针刺促排卵治疗内分泌失调性不孕临床观察[J].中国医学理论与实践,2003；23(4)：524
[26]常静玲.针灸治疗女性不孕症32例[J].上海针灸杂志,1998；17(1)：26
[27]张继红,张慧岭,赵藏朵,等.补肾化浊法药针并用治疗免疫性不孕23例[J].陕西中医,2005；26(10)：1024～1025
[28]唐晔,贾淑华,张庆蔚.针药并用治疗抗精子抗体阳性不孕症40例[J].中国民间疗法,2001；9(9)：9-10
[29]李忠仁.实验针灸学[M],北京；中国中医药出版社,2003,2：198～200,203
[30]周雄元,王跃华.针刺调节免疫功能的机理研究进展[J].湖北中医杂志,2004；26(7)：56
[31]唐照亮,宋小鸽,侯正明,等.大鼠海马内微量注射6羟多巴胺对艾灸抗炎免疫作用的影响[J].针刺研究,2000；25(3)：184
[32]赵建础.针刺调节免疫反应途径的初步研究[J].针刺研究,1997；22(1)：91
[33]张海峡,贾成文.针灸对红细胞免疫的研究进展[J].陕西中医学院学报,2004；27(1)：22～24
[34]杨会芹,张联科.针灸对红细胞免疫功能调节的研究[J].陕西中医学院学报,2004；27(5)：76～77
[35]严伟星,郑震,王健鹤.β—内啡肽对细胞免疫功能的调节[J].南通医学院学报,1994；14(2)：134
[36]何燕萍,唐纯志,梁国珍等.针药结合疗法对免疫性不育大鼠血清抗精子抗体的影响.广州中医药大学学报,2001；18(2)：142
[37]伦新.针刺治疗对男性免疫性不育症T淋巴细胞亚群分类的影响[J].北京中医药大学学报,2004；27(4)：90
[38]何燕萍,宋阳,张小洪.归肾丸结合针刺对免疫性不育症的临床作用及对精浆酸性磷酸酶的影响[J].广州中医药大学学报,2007；24(5)：361
[39]马朱红,刘心莲,尹淑英,等.针灸对类风湿性关节炎患者免疫一神经—内分泌系统影响的临床研究[J].中国针灸,1997；16(9)：529
[40]李辉,牟晓泓,翟景慧.督脉腧穴电针对佐剂性关节炎大鼠IL-2影响的实验观察[J].针刺研究,2004；29(1)：54
[41]Saji F,Nogoro T,Ohashi K etal clinical evaluation of the enzyme-linked immunosorbent assay kitfor antisperm antibodies.Fertil steril,1988,50:644
[42]Korolkova Iv,Nikolaova MA,Boghedomov VA,etal Free radical generation in ejaculate samples from infertile patients [J].Bull Exp Biol Med,2001,13(6):555-557
[43]Myogo K,Yamano S,Nakagawa K,etal Sperm-immobilizing antibodies block capacitation in human spermatozoa [J].Arch Androl,2001,47(2):135-142
[44]王浩飞,向祖琼,王益鑫.60KD的精子膜抗原对抗精子抗体阳性精子顶体酶活性的影响[J].中华男科学杂志,2004；18(2)：15～18
[45]Shibahara H. (?)igeta M. etal.Diversity of the blocking effects of antisperm antibodies on fertilization in human and mouse.Hum Report,1996,11:2595-2599
[46]潘柏轩,龙峰.不孕不育与多种自身抗体的相关性研究[J].中华男科学杂志,2006；12(11)：1039
[47]朱长庚,刘庆莹,魏瑛,等.大鼠中枢神经元内免疫神经内分泌物质的共存[J].解剖学报,1999；30(2)：100
[48]胡电,戎霖.内源性阿片肽与子宫内膜异位不孕[J].实用妇产科杂志,1993；9(6)：308
[49]倪鑫,林葆成,王成海,等.β—内啡肽对大鼠免疫功能的影响[J].中国免疫学杂志,1993；9(1)：16
[50]Carcia I,Perez-castillo A,Moreno J,etal.β-endorphin inhibits interleukin-2 release and expression of interleukin-2 receptors in concanavalin A-stimulated splenic lymphocytes.Lymphokine Cytoline Res,1992;11:339-345
[51]王玉秀,童昭岗,胡海青.内源性阿片肽系统免疫调节功能的研究进展[J].中国临床康复,2003；7(10)：1559
[52]孙志扬,过宗南,赵孟尧.β—内啡肽对正常人红细胞免疫功能的调控实验研究[J].上海免疫学杂志,1994；14(4)：200
[53]RhanNA.FernereF,Deschaux P Serotonin—inducedcalctum via 5-HT1a receptors in Human Leukeima(R562)cells Cellul Immunol,1995.165:148
[54]LabergSCrikshankwwBeerDj.etalSecretionofIL6(lym. Phocytechemottractant factor)from serotonin—stimulated CD8+Ttells in vitro J Immunol.1996,156:310
[55]张清泉,彭聿平,邱一华,等.5—羟色胺与体液免疫应答的相互作用[J].南通医学院学报,1995；15(4)：504～505
[56]高娜,王阿敬,杨渝珍,等.阿片受体介导大鼠海马内脑啡肽对细胞免疫功能的调节[J].生理学报,1999；51(5)：106～109
[57]李晓玉.甲硫氨酸脑啡肽的免疫调节作用[J].中国药理学报,1998；19(1)：5～6
[58]严伟星,王健鹤,张清皋.纹状体中亮一脑啡肽在针刺调节免疫功能中的作用[J].生理学报,1991；43(5)：451～456
[59]张龚,陈丽萍,任芬若,等.不明原因不孕妇女宫颈粘液中抗精子抗体、IL-2和IFN-r 的测定[J].中国免疫学杂志,2000；16(2)：624
[60]余琦.女性慢性生殖道炎症与抗精子免疫的相关研究.广州中医药大学硕士学位论文,(2002届)：21
[61]金忆.不明原因不孕患者宫颈粘液白细胞介素2及肿瘤坏死因子a[J].中华妇产科杂志,1999；34(8)：499
[62]侯淑萍.不孕症患者血清TNF、IL—2、E2、FSH、LH、P、PRL水平的变化[J].淮海医药,2002；20(4)：272
[63]Tezawala BU. JohnsonP. ReesRC. Inhibition of pregnancy viability in mice following IL—2 adrninistration[J]. Immunology,1989;67:115
[64]Beckman.Js,KeoppenolWH,Nitricoxide,superoxide,andperoxynitrite:thegood,thebad,t heugly,AmJPhysiol,1996;27(cellphysio140):1424-1437
[65]Chwaliszk,Buhimschi,Garfield RE.edtal Role of nitric oxide in obstetrics,prenat Neonat Med,1996;1(3):292-329
[66]RosselliM.Nitricoxideandreproduction.HumReprod:1997,3:639-641
[67]解启发.NO在小鼠早期胚胎吸收过程中的作用[J].中国兽医学报,2002；22(1)：90～93
[68]龙振州.医学免疫学[M],北京：人民卫生出版社,1998,2：79
[69]Heas GG. Immunologic infertility:which approach are best? Cotemp Obstet Gynol.1983;22:41
[70]罗颂平,张玉珍,梁国珍,等.免疫性自然流产与免疫性不孕的临床与实验研究[J].中医杂志,1997；38(6)：351～354
[71]Bronson R. Sperm antisbodies:Their role in infertility. Fertil steril,1984,42: 171
[72]吴爱明,郭霞.清化消抗汤治疗抗精子免疫性不孕症60例[J].河南中医,2007；27(3)：50
[73]王丽,黄萍,黄晓燕,等.1020例免疫性不孕患者的治疗结果分析[J].中华妇产科杂志,1999；34(4)：234～235
[74]刘继红,章咏裳,邓荣进,等.抗精子抗体阳性不育患者外周血淋巴细胞亚群检测及其意义[J].中国男科学杂志,1990；4(4)：200
[75]赵武述等.现代临床免疫学[M],北京：人民军医出版社,1994,1：98
[76]刘静君.种子转阴颗拉治疗免疫性不孕症(抗精子抗阳性)的临床与实验研究.山东中医药大学硕士学位论文,(2002届)：29
[77]赵蓉,杨金洲,郁美娟,等.针刺对放化疗患者T细胞及其亚群影响[J].上海针灸杂志,1994；13(6)：253
[78]陈雄华,刘又香.针灸足三里、关元对阳虚大鼠免疫功能影响的比较研究[J].中 国针灸,2000；20(9)：555
[79]谭敬书,徐绍勤,李凡成.穴位敷贴治疗常年性变态反应性鼻炎的临床研究[J].湖南中医学院学报,1994；14(2)：56～57
[80]赵宁侠,高巍,黄裕新,等.电针足三里穴对免疫抑制大鼠细胞免疫影响[J].针刺研究,2002；27(1)：56.
[81]魏赞美,庄肩岚.针刺足三里治疗白细胞减少症及对免疫功能观察[J].上海针灸杂志,1996；15(6)：12
[82]邱艳明.电针对抑郁模型大鼠单胺类神经递质影响的实验研究.北京中医药大学,(2002届)：74
[83]金光亮,周东丰,苏晶.电针对慢性应激抑郁模型大鼠脑单胺类神经递质的影响[J].中华精神科杂志,1999；32(2)：220
[84]吴绪平,冯异,刘又香,等.针刺对急性脑梗塞大鼠血浆cAMP与β—EP的影响[J].中国针灸,2002；22(7)：486
[85]丁建江,张磊.头穴透刺治疗脑梗塞及对血浆中β—内啡肽含量的影响[J].时珍国医国药,2007；18(10)：2536
[86]李辉,李晓泓,张露芬,等.电针对佐剂性关节炎大鼠下丘脑CR H、IL—2、β—EP含量的影响[J].中国针灸,2005；25(11)：795
[87]杨俊,黄祖汉.针刺对大鼠β—内啡肽含量的影响[J].针灸学报,1992；16(6)：31
[88]曲丽芳,李明权,白华,等.中药复方对实验性老年高脂血症红细胞免疫粘附功能和循环免疫复合物的作用[J].中国免疫学杂志,1996；12(2)：107
[89]国家中医药管理局.中华本草,上海：上海科技出版社,1999,1：667
[90]蔡定芳,沈自尹,张玲娟,等.右归饮对皮质酮大鼠细胞免疫及细胞因子的影响[J].中国免疫学杂志,1995；11(4)：248
[91]钟雪梅,周灵,杨家林,等.补经合剂促卵泡发育和排卵的实验研究[J].江苏中医,1998；19(3)：45
[92]沈皓,蔡德培.补肾中药对下丘脑—垂体促性腺机能的影响[J].中西医结合学报,2004；1(2)：53
[93]张炜,蔡德培.补肾中药对雌性青春期大鼠垂体GnHR受体mRNA及其受体蛋白表达的影响[J].中国中西医结合杂志,2003；23(2)：120