Objective

2

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

2

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

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

2

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

2

in the plasma and E

2

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(

P

<

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 (

P

<

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

2

contents. The 6-keto-PGF1α content increased

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

P

<

0.05). The high

Carthami Flos

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

P

<

0.05). In the HSYA group

the writhing response decreased

the uterine and ovarian indices decreased

the PGE2

PGF2α

and TXB

2

contents decreased

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

P

<

0.05). The serum E

2

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 (

P

<

0.05).

Conclusion

2

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.