月经病寒凝血瘀证大鼠卵巢HO/CO变化及与生殖内分泌的关系

目的:观察月经病寒凝血瘀模型大鼠血红素氧合酶/一氧化碳(HO/CO)系统的变化及与生殖内分泌的关系,探讨HO/CO系统在寒凝血瘀妇科疾病的发生中的作用。方法:20只大鼠,随机分为模型组和正常组。模型组大鼠置于0℃-1℃冰水中20分钟,每日1次,连续2周,2周后断头处死,取血,采用放射免疫法测定血清E2、P活性,采用血红蛋白结合及联二亚硫酸盐还原法测定血浆CO水平;剖取卵巢,采用免疫组织化学SP法检测卵巢组织HO-1、HO-2蛋白表达;采用原位杂交法检测卵巢组织HO-1、HO-2 mRNA表达。结果:模型组大鼠血清E2、P下降,血浆COHb活性降低,与正常组比较有统计学意义。模型组大鼠卵巢颗粒细胞、黄体细胞HO-1、HO-2蛋白及mRNA表达均下降,与正常组比较差异有统计学意义。并且血浆COHb活性、卵巢组织HO-1、HO-2表达与血清E2、P呈正相关。结论:寒凝血瘀大鼠HO/CO系统抑制,生殖内分泌紊乱,卵巢功能失常从而引发月经病。

Change of syndrome of cold model rat of blood stasis due to accumulation of cold in HO/CO system and the relations with reproductive endocrine

Objective: To observe the change of syndrome of cold model rat of blood stasis due to accumulation of cold in HO / CO system and the relations with reproductive endocrine,and to explore the role of place on Gynecological disease.Methods: 20 rats were divided into model group and normal group randomly,10 rats in each group.The rats in model group were placed in water at 0~1℃for 20 minutes,one time a day,guillotined and draw the blood two weeks later.The E2,P activity in serum was detected by the method of radioimmunoassay.The CO activity was detected by the methods of protein binding and dithionite reduction;the protein expression of HO-1、HO-2 in ovary was determined by Immunohistochemistry analysis;in situ hybridization histochemistry analysis was used to reveal the mRNA expression of HO-1、HO-2 in ovary.Result: Compared with the normal group,The activity of plasma COHb were significantly decreased.And the protein expression and mRNA expression of HO-1 and HO-2 in ovarian granulosa cells and luteal cells were decreased in the model group.And the plasma COHb activity,HO-1,HO-2 expression in ovarian tissue were positively related to the E2,P activity in serum.Conclusion: The HO/CO system were inhibition in cold model rats,reproductive endocrine disorders,ovarian dysfunction causing menstrual disease.