1.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
2.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
3.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
4.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
5.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
6.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
7.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
8.Study on effect and mechanisms of Carthami Flos water extract and hydroxysafflower yellow A on primary dysmenorrhea rats with cold coagulation and blood stasis
Ran WANG ; Ying SUN ; Boyang JIAO ; Chun LI ; Jun LI ; Pengfei TU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1397-1407
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 B2(TXB2)and 6-keto-prostaglondin F1α(6-keto-PGF1α)in plasma,and estradiol(E2)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.TXB2 in the plasma and E2 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 indicesand PGE2 and TXB2 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 TXB2 contents decreased,and GnRH-R and FSH-R protein expression decreased in the uterus(P<0.05).The serum E2 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 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.
9.Mechanism of Intervening with Diarrhea-predominant Irritable Bowel Syndrome in Rats with Spleen Deficiency by Xingpi Capsules Through Regulating 5-HT-RhoA/ROCK2 Pathway
Gang WANG ; Lingwen CUI ; Xiangning LIU ; Rongxin ZHU ; Mingyue HUANG ; Ying SUN ; Boyang JIAO ; Ran WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):60-69
ObjectiveTo investigate the efficacy of Xingpi capsules (XPC) in treating diarrhea-predominant irritable bowel syndrome (IBS-D) with spleen deficiency and elucidate its potential molecular mechanisms. MethodsA rat model of IBS-D with spleen deficiency was established by administering senna leaf in combination with restrained stress and swimming fatigue for 14 d. Ten specific pathogen free (SPF)-grade healthy rats were used as the normal control group. After successful modeling, SPF-grade rats were randomly divided into a model group, a pinaverium bromide group (1.5 mg·kg-1), and low- and high-dose XPC groups (0.135 and 0.54 g·kg-1), with 10 rats in each group. Rats in the normal control group and the model group were given distilled water by gavage, while the remaining groups were administered corresponding drug solutions by gavage once a day for 14 consecutive days. The rat body weights and fecal condition were observed every day, and the Bristol score was recorded. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of 5-hydroxytryptamine (5-HT) in serum and colon tissue. Transmission electron microscopy was used to observe the microvilli and tight junctions in the colon. The integrity of the colonic barrier, intestinal motility, and expression of related pathway proteins were evaluated by hematoxylin-eosin (HE) staining, immunohistochemistry, and Western blot. ResultsCompared with those in the normal control group, rats in the model group showed a significantly decreased body weight and increased diarrhea rate, diarrhea grade, and Bristol score (P<0.01). HE staining revealed incomplete colonic mucosa in the model group, with evident congestion and edema observed. Electron microscopy results indicated decreased density and integrity of the colonic barrier, shedding and disappearance of microvilli, and significant widening of tight junctions. The expression levels of colonic tight junction proteins Occludin and Claudin-5 were downregulated (P<0.01), and the levels of 5-HT in serum and colon tissue were elevated (P<0.01). The small intestine propulsion rate significantly increased (P<0.01), and the expression of contractile proteins Ras homolog family member A (RhoA) and Rho-associated coiled-coil containing protein kinase 2 (ROCK2) in colon and phosphorylation of myosin light chain (MLC20) were upregulated (P<0.01). Compared with the model group, the treatment groups showed alleviated diarrhea, diarrhea-associated symptoms, and pathological manifestations of colon tissue to varying degrees. Specifically, high-dose XPC exhibited effectively relieved diarrhea, promoted recovery of colonic mucosal structure, significantly reduced congestion and edema, upregulated expression of Occludin and Claudin-5 (P<0.01), decreased levels of 5-HT in serum and colon tissue (P<0.05,P<0.01), significantly slowed small intestine propulsion rate (P<0.01), and significantly downregulated expression of contractile proteins RhoA and ROCK2 in colon and phosphorylation of MLC20 (P<0.05,P<0.01). ConclusionXPC effectively alleviates symptoms of spleen deficiency and diarrhea and regulates the secretion of brain-gut peptide. The characteristics of XPC are mainly manifested in alleviating IBS-D with spleen deficiency from the aspects of protecting intestinal mucosa and inhibiting smooth muscle contraction, and the mechanism is closely related to the regulation of the 5-HT-RhoA/ROCK2 pathway expression.