Study On The Pharmacodynamics And Mechanism Of Si-ni Decoction In The Treatment Of Dysmenorrhea With Cold Coagulation And Blood Stasis

Objective:Through si-ni decoction in the treatment of blood stasis type of dysmenorrhea haemorrheological nature in the rat model of pharmacodynamics research,clear si-ni decoction on blood stasis type of dysmenorrhea haemorrheological nature of the validity of the model of rats,then the network pharmacology analysis method is adopted to predict its action mechanism,and the predicted results are preliminary experimental verification,to explore si-ni decoction in the treatment of blood stasis type of dysmenorrhea haemorrheological nature of effect mechanism.Methods:1.The pharmacodynamic study of si-ni decoction on the rat model of cold coagulation and blood stasis and dystrorrhagiaFemale Wistar rats were randomly divided into 7 groups:Blank group,model group,positive chemical medicine control group(ibuprofen 0.133 g·kg^-1),traditional Chinese medicine（TCM）positive control group(herba leonuri particle 5 g·kg^-1),si-ni decoction of high,middle,low dose group(the equivalent of crude drugs 16 g·kg^-1,8 g·kg^-1,4 g·kg^-1),copy the model of blood stasis type of dysmenorrhea haemorrheological nature,in building3 days lavage（ig）give relevant drugs,11 days in a row,groups of rats after the last delivery,intraperitoneal injection of oxytocin(2 U·100g^-1),to observe rat body torsion latent period and body torsion reaction times within 30 minutes;The quality of rat uterus,ovary and kidney were measured,the index of viscera was calculated,and the changes of tissue morphology were observed by HE staining.Total blood viscosity,plasma viscosity and hematocrit in plasma were measured.The contents of nitric oxide（NO）,prostaglandin F2α（PGF2α）,prostaglandin E2（PGE2）,estrogen（E2）,progesterone（PROG）,luteinizing hormone（LH）,follicle-stimulating hormone（FSH）and 5-hydroxytryptamine（5-HT）in uterine tissue were detected by ELISA.2.Study on the mechanism of si-ni decoction based on network pharmacology in treating dysmenorrhea of cold coagulation and blood stasisNetwork pharmacology method and in combination with related literature review,collect si-ni decoction prescription medicine aconite root,dried ginger,the chemical composition of licorice,using TCMSP respectively in Chinese native medicine ingredient database"aconite root,dried ginger,licorice"as the keyword search and collect every flavour Chinese medicine related to chemical composition,and based on TCMSP pharmacokinetic parameters in the database（ADME）by oral use of absorbance（OB）30%or higher,classes,medicinal（DL）four decoction potential acuity 0.18 for screening of the active ingredient,The Swiss Target Prediction server was used to predict the potential targets of the active components of si-ni decoction based on the ligand structure.And through DrugBank,DisGeNET,TTD and other databases to“Dysmenorrhea,Primary Dysmenorrhea”as the retrieval conditions to collect disease targets related to Primary Dysmenorrhea;For the target from si-ni decoction active ingredient prediction map with primary dysmenorrhea related targets,screening si-ni decoction treatment of primary dysmenorrhea may work target,target through DAVID website for the enrichment of gene ontology（GO）function analysis and KEGG signal pathway analysis,build si-ni decoction active ingredient-primary dysmenorrhea disease targets-metabolic pathway network diagram,discusses si-ni decoction in the treatment of blood stasis type of dysmenorrhea haemorrheological nature of effect mechanism.3.Verified the mechanism of si-ni Decoction in treating dysmenorrhea caused by cold coagulation and blood stasisWestern blotting was used to detect the expression levels of prostaglanin G/H synthase 1（PTGS1）,prostaglanin G/H synthase 2（PTGS2）in uterine tissue.Results:1.Study on the pharmacodynamic effect of si-ni decoction on the rat model of cold coagulation and blood stasis and dysmenorrheaSi-ni decoction can relieve pain in the rat model with blood stasis type of dysmenorrhea haemorrheological nature,reduce the body torsion times,decrease the whole blood viscosity and plasma viscosity,erythrocyte deposited,reduce serum 5-HT,E2,P,LH,FSH and uterus homogenate PGF2αlevels,improve the level of PGE2 in the uterus homogenate,NO,and improve the uterus,ovarian tissue pathological changes.Speculated that the treatment of blood stasis dysmenorrhea haemorrheological nature of effect mechanism may be related to improve blood viscosity,increase blood flow,influence the uterus tissue NO,PGF2α,PGE2 and 5-HT,E2,P in serum,LH,LSH hormone level,at the same time,E2,P,LH,LSH hormone and the hypothalamus-pituitary-ovarian axis（HPOA）is closely related to the neuroendocrine system,si-ni decoction could prompt by adjusting the adjustingPGE2,PGF2α,the NO content in the uterus tissue and the hypothalamus-pituitary-ovarian axis neuroendocrine system related hormone levels achieve alleviate the effect of blood stasis type of dysmenorrhea haemorrheological nature.2.Study on the network pharmacological mechanism of si-ni decoction in treating dysmenorrhea of cold coagulation and blood stasisReceived through the analysis of the network pharmacology of aconite,dried ginger,licorice potential active ingredient,a total of 106 related to dysmenorrhea disease targets27,si-ni decoction treatment of primary dysmenorrhea may target 27,KEGG pathway enrichment analysis found that the mechanism of action of si-ni decoction treatment of primary dysmenorrhea with arachidonic acid metabolism,linoleic acid metabolism,chemical carcinogens,steroid hormone biosynthesis and other signaling pathways.3.Verified the mechanism of Sini decoction in treating dysmenorrhea caused by cold coagulation and blood stasisWestern blotting results showed that:PTGS1 protein expression in the uterus of the model group was significantly decreased（p<0.05）,and PTGS2 protein expression was significantly decreased（p<0.01）.By upregulating PTGS1 and PTGS2 protein expression（p<0.05）,herba nooyi granules could alleviate dysmenorrhea caused by cold coagulation and blood stasis,while sini decoction group had no significant effect on PTGS1 and PTGS2 protein expression.It is suggested that the key target of si-ni decoction for its pharmacological effect is not PTGS1 and PTGS2,but other proteins in the arachidonic acid signaling pathway.Conclusion:Si-ni decoction has therapeutic effect on dysmenorrhea caused by cold coagulation and blood stasis in rats,and this effect may be related to regulating arachidonic acid metabolism,chemical carcinogen,linoleic acid metabolism,steroid hormone biosynthesis synaptic,retinol metabolism,drug metabolism,serotonin-cytochrome P450,cytochrome P450 of exogenous biological metabolism,ABC transporters,ovarian steroidogenesis,bile secretion,metabolic pathway 12 signaling pathways.