Study on effect and mechanisms of <italic style="font-style: italic">Carthami Flos</italic> water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis

WANG Ran; SUN Ying; JIAO Boyang; LI Chun; LI Jun; TU Pengfei

doi:10.3969/j.issn.1006-2157.2024.10.010

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

Experimental Study | 更新时间：2024-10-29

- Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
- Journal of Beijing University of Traditional Chinese Medicine Vol. 47, Issue 10, Pages: 1397-1407(2024)
- 作者机构：
  
  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
- 作者简介：
  
  Prof. TU Pengfei, Ph.D., Doctoral Supervisor. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Science, Peking University, No. 38, Xueyuan Road, Haidian District, Beijing 100191. E-mail: pengfeitu@163.com
- 基金信息：
  
  National Key R&D Program of China(2022YFC3501602)
- DOI：10.3969/j.issn.1006-2157.2024.10.010
  CLC： R285.5
- Received：15 May 2024，
  
  Published：30 October 2024
- 稿件说明：
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WANG Ran, SUN Ying, JIAO Boyang, et al. Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(10): 1397-1407.
DOI：

WANG Ran, SUN Ying, JIAO Boyang, et al. Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(10): 1397-1407. DOI： 10.3969/j.issn.1006-2157.2024.10.010.

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

(TXB

)、6-酮-前列腺素F1α(6-keto-PGF1α)含量；放射免疫法检测大鼠血清中雌二醇(E

)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

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(

0.05)。与模型组比较，红花水提物低剂量组子宫指数降低(

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

降低，6-keto-PGF1α升高，子宫GnRH-R蛋白表达降低(

0.05)；红花水提物高剂量组PGF2α降低，子宫组织FSH-R表达降低(

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

含量降低，子宫GnRH-R、FSH-R蛋白表达降低(

0.05)；红花水提物各剂量及羟基红花黄色素A组血清E

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(

0.05)。

结论

红花及羟基红花黄色素A能够改善原发性痛经寒凝血瘀证大鼠的病理状态，其机制可能与调控下丘脑－垂体－卵巢轴有关。

Abstract

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 enzyme-linked immunosorbent assay and radioimmunoassay were used to detect the prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) contents in the uterus

thromboxane B

(TXB

) and 6-keto-prostaglondin F1α (6-keto-PGF1α) in plasma

and estradiol (E

) 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 PGF2α in the uterus. TXB

in the plasma and E

in the serum were also evaluated. Additionally

6-keto-PGF1α decreased

and COX-2

GnRH-R

and FSH-R expression in the uterus and FSH-R expression in the ovaries also increased(

0.05). The morphology of the uterine tissue was disordered. Compared with the model group

the low

Carthami Flos

water extract dose group showed a decrease in uterine index (

0.05). In the medium and high

Carthami Flos

water extract dose groups

the writhing response decreased

as did the uterine and ovarian indices and PGE2 and TXB

contents. The 6-keto-PGF1α content increased

whereas the GnRH-R protein expression in the uterus decreased (

0.05). The high

Carthami Flos

water extract dose group also showed a decrease in PGF2α and FSH-R protein expression in the uterus (

0.05). In the HSYA group

the writhing response decreased

the uterine and ovarian indices decreased

the PGE2

PGF2α

and TXB

contents decreased

and GnRH-R and FSH-R protein expression decreased in the uterus(

0.05). The serum E

levels of the groups that received

Carthami Flos

water extract at various doses and those of the HSYA group were reduced

and the uterine morphology was improved. COX-2 expression in the uterus and FSH-R protein expression in the ovaries were also reduced (

0.05).

Conclusion

Carthami Flos

water extract and HSYA can improve the pathological state of primary dysmenorrhea rats with cold coagulation and blood stasis. Its mechanism may be related to regulating the hypothalamic-pituitary-ovary axis.

关键词

Keywords

references

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

DOI：10.3969/j.issn.1006-2157.2024.10.010

摘要

Abstract

关键词

Keywords

references