赤黄平阳汤对雌性大鼠抗精子抗体抑制作用的实验研究

【作者】 张倩；

【导师】 张丽君；

【作者基本信息】 湖北中医学院 ， 中医妇科学， 2007， 硕士

【摘要】 目的：本课题旨在从理论上阐明“毒邪”是抗精子抗体产生的病因病机，采用同种异体人工主动免疫法建立雌性大鼠抗精子抗体阳性动物模型，观察赤黄平阳汤对模型大鼠的抗精子抗体吸光度(OD)值、妊娠率、胚胎数、各脏器重量及子宫、卵巢组织学的影响，证实以清热解毒、凉血活血组方的赤黄平阳汤对抗精子抗体阳性的抑制作用，从而为临床的广泛应用提供科学依据。方法：本实验选用2-3月龄SD雌鼠50只，从中随机取10只作为正常对照组，余40只大鼠双侧腹股沟皮下淋巴结周围注射抗原进行免疫，最多经5次免疫后，所有大鼠血清AsAb为阳性。将40只抗精子抗体阳性大鼠按随机数字表法随机分为：模型组、赤黄平阳汤组、知柏地黄汤组、醋酸泼尼松组，其中正常组及模型组灌服生理盐水，余治疗组分别灌胃给药。两周后，各治疗组雌鼠眶内静脉丛取血，离心取血清，标记后保存，酶联免疫法(ELISA)统一测定抗精子抗体吸光度(OD)值。同时，以雄与雌为1／3比例配对合笼，于妊娠第18天，断头处死雌鼠，数胚胎数及计算各组受孕率。同时解剖各雌鼠，对肝，脾及双肾分别称重并进行记录。处死雌鼠前再一次检测抗精子抗体吸光度(OD)值。所有雌鼠子宫及卵巢甲醛固定，包埋，切片，观察子宫内膜及卵巢的组织学变化。结果：1．药物对各组大鼠AshbOD值的影响：免疫5次后，40只雌鼠AshbOD为阳性，提示造模成功。且各组AsAbOD值比较无差异；合笼前与模型组比较，醋酸泼尼松组与赤黄平阳汤组均可以明显降低血清AsAbOD值(P＜0.01)，但两者相比无差异。合笼约20天后，与模型组比较，各药物组均表现出降低AshbOD值作用(P＜0.01)，三组比较，赤黄平阳汤组优于知柏地黄汤组与醋酸泼尼松组(P＜0.05)。2．各组的受孕率及平均胚胎数比较：赤黄平阳汤组，知柏地黄汤组及醋酸泼尼松组各自受孕率分别为80％，70％，50％，均大于模型组的受孕率40％，拟做行×列表资料的X~2检验，但由于本实验的样本例数较少，选用四格表的确切概率法统计，各组受孕率比较无差异。模型组胚胎数明显降低，与正常对照组比较有统计学意义(P＜0.05)，说明抗精子抗体阳性模型雌鼠确实能降低其胚胎数，而赤黄平阳汤组能增加胚胎数，且与模型组比较经统计学处理有显著性意义(P＜0.05)，知柏地黄汤组及醋酸强地松组也能增加胚胎数，但无统计学意义。3．各组间器官重量比较，其中模型组、醋酸泼尼松组脾脏的重量与正常对照组比较有明显减轻(P＜0.05，P＜0.01)。肾脏与肝脏重量各组比较无统计学意义。4．各组子宫、卵巢组织学观察未发现有明显变化。结论：赤黄平阳汤能有效降低抗精子抗体阳性雌鼠AsAbOD值，增加平均胚胎数，且未发现对各脏器重量有减轻作用。更多还原

【Abstract】 ObjectiveIn order to clarify that "duxie" is the basic pathogenesis of Anti-sperm Antibody, thispaper estabilished anti-sperm antibody masculine animal models with the female rats, andused by active immunization of homologous modified character method,studied the effectsof Chihuangpingyangtang on OD of anti-sperm antibody, pregnancy rate,the number ofembryo,the weight of organ and observed pathologic changes of uterus, ovary to provedrestraint role of Chihuangpingyangtang and offered gist for clinical medicine.MethodsTotal 50 female rats at the age of 2 to 3 months,ten rats were randomly selected asnormal control group,another 40 rats were separately injected antigen to get immunearound subcutaneous lymph nodes of bilateral inguinal. After immunizations for fivetimes,all of female rats’serum was positive and these rats were randomly divided intomodel group by random number table method, Prednisone group, Phihuanpingyangtanggroup, Zhibaidihuangtang group,which were separately given oral medicine.Normalgroup and model group were administered with physiologic saline orally. After two weeks,blood was adopted from eyes,separated serum and detected anti-sperm antibody of serumby enzyme linked immune sorbet assay. At the same time,to collect group statistics of thenumber of embryos and pregnancy rate after 18 days when 3 male rats were added to thefemale in each group. To explore the histology changes of rats’ovaries and uteruses andrecord the weight of liver, spleen,kidney when dissected rats.Results1.The effect Of AsAb OD on medicine:after immunizations for five times,fortyfemale rats’serum was positive and there is no difference about AsAb OD.animal modelswas estabilished successfully.Before mating,AsAb OD of Chihuanpingyangtang andPrednisone group was lower than model group.After about twenty days, AsAb OD ofeach group were more lower than model group, Chihuangpingyangtang group had bettereffect than Zhibaidihuangtang group and prednisone group.2.The number of embryo and pregnancy rate:The pregnancy, rate of Chihuanpingyangtang,zhibaidihuangtang and prednisone group is higher than modelgroup. There is no statistics because of specimen. The number of embryo of model groupare lower than the normal group. The number of embryo Chihuangpingyangtang groupmuch more than model group.3.The weight of organ:The weight of spleen of model and prednisone group is lowerthan normal group.there are no difference each group about the weight of liver and kidney.4.There is no obvious change of uterus and ovary.ConclusionsChihuangpingyangtang could decrease AsAb OD and increase the number ofembryo while the weight of organ don’t be drop.更多还原