红花水提物及羟基红花黄色素A对原发性痛经寒凝血瘀证大鼠的作用及机制研究  

Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis

作　　者：王然[1,2] 孙颖 焦伯阳 李春 李军 屠鹏飞[4] WANG Ran;SUN Ying;JIAO Boyang;LI Chun;LI Jun;TU Pengfei(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Modern Research Center forTraditional Chinese Medicine,Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China;School of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China;State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University,Beijing 100191,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]北京中医药大学北京中医药研究院中药现代研究中心 [3]北京中医药大学中医学院 [4]北京大学药学院天然药物及仿生药物国家重点实验室

出　　处：《北京中医药大学学报》2024年第10期1397-1407,共11页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家重点研发计划项目(No.2022YFC3501602)。

摘　　要：目的探讨红花水提物及其主要单体成分羟基红花黄色素A改善大鼠原发性痛经寒凝血瘀证的药效及调控机制。方法取42只6~8周龄雌性SPF级SD大鼠,按随机数字表法分为空白组、模型组、布洛芬组(0.04 g/kg)、羟基红花黄色素A组(0.01 g/kg)和红花水提物低(0.06 g/kg)、中(0.20 g/kg)、高(0.40 g/kg)剂量组,每组6只。采用冰水浴刺激联合苯甲酸雌二醇及缩宫素建立大鼠模型。造模第7天开始灌胃相应药物,连续6 d。干预结束后,进行扭体反应测试;称量大鼠体质量及子宫、卵巢质量,计算脏器指数;酶联免疫吸附测定(ELISA)检测大鼠子宫前列腺素E2(PGE2)、前列腺素F2α(PGF2α)含量,血浆中血栓素B_(2)(TXB_(2))、6-酮-前列腺素F1α(6-ketoPGF1α)含量;放射免疫法检测大鼠血清中雌二醇(E2)含量;苏木精-伊红染色法检测子宫组织病理状态;免疫组织化学法检测子宫组织环氧合酶(COX-2)表达;免疫荧光法检测大鼠卵巢组织卵泡刺激素受体(FSH-R)表达;蛋白质印迹法检测大鼠子宫促性腺激素释放激素受体(GnRH-R)、FSH-R表达。结果与空白组比较,模型组大鼠子宫、卵巢指数增加,子宫PGE2、PGF2α升高,血浆中TXB_(2)升高、6-keto-PGF1α降低,血清中E2升高,可见子宫组织形态紊乱,子宫COX-2表达升高,卵巢FSH-R表达升高,子宫GnRH-R、FSH-R表达升高(P<0.05)。与模型组比较,红花水提物低剂量组子宫指数降低(P<0.05);红花水提物中、高剂量组扭体次数减少,子宫、卵巢指数降低,PGE2、TXB_(2)降低,6-keto-PGF1α升高,子宫GnRH-R蛋白表达降低(P<0.05);红花水提物高剂量组PGF2α降低,子宫组织FSH-R表达降低(P<0.05);羟基红花黄色素A组大鼠扭体次数减少,子宫、卵巢指数降低,PGE2、PGF2α、TXB_(2)含量降低,子宫GnRH-R、FSH-R蛋白表达降低(P<0.05);红花水提物各剂量及羟基红花黄色素A组血清E_(2)含量降低,子宫形态得到改善,子宫COX-2、卵巢FSH-R蛋白表达降低(P<0.Objective To explore the pharmacological effects and regulatory mechanisms of Carthami Flos water extract and its main constituent,hydroxysafflower yellow A(HSYA),on primary dysmenorrhea rats with cold coagulation and blood stasis.Methods Forty-two female specific pathogen-free grade rats aged 6-8 weeks were divided into blank,model,HSYA(0.01 g/kg),ibuprofen(0.04 g/kg),and low(0.06 g/kg),medium(0.20 g/kg),and high(0.40 g/kg)Carthami Flos water extract dose groups using the random number table method,with six rats per group.A rat model was established using ice water bath stimulation combined with estradiol benzoate and oxytocin.Continuous gavage was administered for 6 days from the seventh day of modeling.After the intervention,the writhing reaction test was conducted.The rats,uteri,and ovaries were weighed to calculate the organ index.An enzymelinked immunosorbent assay and radioimmunoassay were used to detect the prostaglandin E2(PGE2)and prostaglandin F2α(PGF2α)contents in the uterus,thromboxane B_(2)(TXB_(2))and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E_(2))in the serum.Hematoxylin and eosin staining were used to detect the pathological changes in uterine tissue.Immunohistochemistry was used to determine cyclooxygenase-2(COX-2)expression in uterine tissue,whereas immunofluorescence was used to measure follicle-stimulating hormone receptor(FSH-R)expression in ovarian tissue.Western blotting was used to detect gonadotropin-releasing hormone receptor(GnRH-R)and FSH-R expression in uterine tissue.Results Compared with the blank group,the rats in the model group exhibited an increase in uterine and ovarian indices and increased PGE2 and PGF2αin the uterus.TXB_(2) in the plasma and E2 in the serum were also evaluated.Additionally,6-keto-PGF1αdecreased,and COX-2,GnRH-R,and FSH-R expression in the uterus and FSH-R expression in the ovaries also increased(P<0.05).The morphology of the uterine tissue was disordered.Compared with the model group,the low Carthami Flos water extract dose group

关 键 词：红花 羟基红花黄色素A 原发性痛经 寒凝血瘀证 下丘脑-垂体-卵巢轴 大鼠 

分 类 号：R285.5[医药卫生—中药学]