妇科虚寒证大鼠模型的创建

Establishment of Gynecological Asthenia Cold Syndrome Model

目的模拟妇科虚寒证发生的病因,依据"寒邪伤阳"的中医学理论,创建妇科虚寒证模型。方法将SD雌性大鼠按随机数字表法分为模型组和正常组,每组20只。模型组置于0～1℃冰水中并放入4℃冰箱中,2次/天,20min/次,共30天。每天测定大鼠体温及观察动情周期的变化,当大鼠体温降低(与正常组之间比较差异有统计学意义),阴道细胞涂片显示动情周期紊乱时,显示造模成功。择动情间期处死大鼠,检测血液生殖内分泌激素[血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、睾酮(T)、孕酮(P)]、甲状腺功能[血清总三碘甲状腺原氨酸(T3)、总甲状腺素(T4)、促甲状腺素(TSH)]、肾上腺功能[血浆促肾上腺皮质激素(ACTH)、皮质醇(Cor)]、细胞免疫功能[血清白细胞介素-2(IL-2)]、能量检测[血浆环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)、cAMP/cGMP比值,乳酸脱氢酶(LDH)]及胸腺、脾脏、子宫、卵巢指数。结果与正常组比较,第30天时模型组动物外观差,被毛无光泽,耳、爪紫暗;精神萎靡,蜷缩少动,反应迟钝,喜扎堆;饮水量、食量减少,体重减轻,体温降低显著,便溏;动情周期、间期延长;血浆cGMP含量、cAMP/cGMP比值、LDH、血清IL-2含量及E2、P、T、T3、LH、TSH均有不同程度的降低,胸腺和卵巢指数显著降低,差异均有统计学意义(均P<0.05)。结论采用延长冰箱冷冻加冰水浸泡时间的方法制作的妇科虚寒证大鼠模型,符合妇科虚寒证的临床特征。

Objective To establish the model of gynecological asthenia cold syndrome by simulating the etiology of gynecological asthenia cold syndrome depending on ＂pathogenic cold impairing yang＂ in Chinese medicine theory. Methods The female SD rats were randomly divided into the model group and the normal group by randomized digital table,20 in each group. Rats in the model group were placed in 0℃-1℃ ice water,and the ice water was placed in 4℃ refrigerator,twice daily,20 min each time,for a total of 30 days. The body temperature was determined and the changes of the estrous cycle were observed every day. When the body temperature decreased（with statistical difference from those of the normal group） ,and vaginal smears showed disordered estrous cycle,the model was successfully established. Rats were sarcrificed during the diestrus period,the correlative indices including reproductive endocrine hormones in blood [follicle-stimulating hormone（FSH） ,luteinizing hormone（LH） ,estradiol（E2） ,testosterone（T） ,and progestone（P） ],the thyroid function [triiodothyronine（T3） ,tetraiodothyronine（T4） ,thyrotropic-stimulating hormone（TSH） ],the adrenal function [plasma adrenocorticotropic hormone（ACTH） and cortisol（Cor） ],the cellular immune function [serum interleukin-2（IL-2） ],and energy tests [including plasma cyclic adenosine monophosphate（cAMP） ,cyclic guanosine monophosphate（cGMP） ,the ratio of cAMP/cGMP,lactate dehydrogenase（LDH） ],and the index of thymus,spleen,uterus,and ovary were detected. Results Compared with the normal group,rats in the model group appeared chill,dim-color hair,purple and dark ears and claw. They were depressed,scrunched,quiet,and clumsy. They liked to stay together. Their water intake and appetite were reduced,body weight lost,body temperature significantly lowered. They passed loose stool. Their estrous cycle and diestrus were prolonged. Their plasma cGMP content,cAMP/cGMP ratio,LDH,serum IL-2 content,E2,P,T,T3,LH,TSH were significantly lowered to some extent. Their thymus index and the ovary index significantly decreased,showing significant difference（P0.05） . Conclusion The rat model of gynecological asthenia cold syndrome prepared by extending the frozen time at refrigerator and ice water immersion was in line with clinical features of gynecological asthenia cold syndrome.