Investigating the effects of Wenjing Decoction combined with Hupo Powder on cold-congealed blood stasis in endometriosis rats,using the NF-κB pathway
[Objective]To explore the effects of Wenjing Decoction combined withHupoPowderon cold-congealed blood stasis endometriosis(EMs)rats based on the nuclear transcription factor-κB(NF-κB)pathway and related proteins.[Methods]Sixty SPF-grade female SD rats were randomly divided into sham-operated group,model group,gestrinone categories(GC),and low,medium,and high dose groups(9.69,19.38 and 38.75 g/kg)of Wenjing Decoction combined with Hupo Powder group,with 10 rats in each group.Except for the sham-operated group,the rat model of EMs with cold-congealed blood stasis was established by using ice-water bath combined with epinephrine hydrochloride and autologous endothelial transplantation in all groups of rats.Sham-operation group with only open abdomen without endometrial transplantation.The sham-operated and model groups were given pure water.The progesterone group was given progesterone(0.25 mg/kg)by gavage twice a week(every Monday and Thursday)dissolved in pure water for four weeks.The group of Wenjing Decoction with Hupo Powder was given low dose of Wenjing Decoction with Hupo Powder(9.69 g/kg),medium dose of Wenjing Decoction with Hupo Powder(19.38 g/kg),and low dose of Wenjing Decoction with Hupo Powder(38.75 g/kg)by gastric gavage for four consecutive weeks,respectively.Serum tumor necrosis factor-α(TNF-α)and interleukin 1β(IL-1β)levels were measured using enzyme-linked immunosorbent assay(ELISA)in each group of rats and the blood rheology(plasma viscosity,erythrocyte aggregation index,erythrocyte rigidity index,erythrocyte deformation index,erythrocyte electrophoresis time),coagulation[prothrombin time(PT),prothrombin time international normalized ratio(PT-INR),partial thromboplastin time(TT),fibrinogen(FIB)]was tested in each group of rats.Detection of NF-κB p65,IkBα,and IKKβ protein expression in endometrial tissues of rats in each group using protein immunoblotting.[Results]Compared with the sham-operated group,the serum TNF-α and IL-1β levels were significantly higher in the model group(P<0.05 or P<0.01),and all the indices of blood rheology were significantly higher(P<0.05 or P<0.01).For coagulation,TT was significantly lower(P<0.05)and FIB were significantly higher(P<0.01).The results of protein expression blotting assay showed that NF-κB p65 and IKKβ were significantly elevated(P<0.01),and the expression of IkBα was significantly reduced(P<0.05).Compared with the model group,the expression of serum TNF-α and IL-1β was significantly reduced in the western medicine and part of traditional Chinese medicine(TCM)dose groups(P<0.05 or P<0.01),as well as all the indexes of blood rheology(P<0.05 or P<0.01).For coagulation,TT were significantly higher(P<0.05 or P<0.01)and FIB were significantly lower(P<0.05 or P<0.01).The results of protein expression blotting assay showed that the expression of NF-κB p65 were decreased(P<0.05 or P<0.01),the expression of IKKβ in the western medicine group and part of TCM dose groups were significantly decreased(P<0.05 or P<0.01),the expression of IkBα in the western medicine and part of TCM dose groups were increased(P<0.05 or P<0.01).[Conclusion]Wenjing Decoction combined with Hupo Powder may shrink the ectopic foci and regulate the development of endometriosis by blocking or inhibiting the activation of the NF-κB signalling pathway and inhibiting the proliferation of ectopic tissues,and it can improve the symptoms of cold agglutination and blood stasis,with a therapeutic efficacy superior to that of pregnenolone.
endometriosisWenjing Decoction combined with Hupo PowderNF-κB signalling pathwaycold-congealed blood stasis