Study On The Mechanism Of Blood-activating And Stasis-dissolving Of Tao Hong Siwu Decoction On Blood Stasis Syndrome Rat Based On Rho/Rocks Signaling Pathway

ObjectiveTo explore the therapeutic effect of Taohong Siwu Detoction (TSD) on model rats withblood stasis syndrome of puerperal disease (BSSPD) and its correlation to variablesrelated to blood clotting promotion, fibrinolytic system and Rho/Rocks pathway. Tostudy the effect of TSD on blood circulation promotion, blood enrichment andmenstrual adjustment, so to provide experimental evidence to explain its clinicalefficacy.Methods1. Therapeutic effect of TSD on rat models with BSSPD. Qualified SD rats wereweighed and encoded after a week of adopting feeding. Two female and one male ratswere arranged together, vaginal suppository and vaginal smear with sperm in the nextmorning were used to determine, if yes, then be the first day of pregnancy. BSSPD ratmodels were copied by fedding with mifepristone and misoprostolon on the7th day ofpregnancy. Six groups were designed, there were control, positive control, model, highdose, medium dose and low dose of TSD. The next day, rats model were administeredfor seven consecutive days. Forty minutes after the last administration, abdominal aorticblood was extracted from anesthetized rats, which would be used to measurehemorheologic and fibrinolytic system indicators. Then the uterine tissue was extractedand one side was paraffin-embedded for uterine pathology morphological observation,and the other side was used to examine uterine microvascular density byimmunohistochemistry method.2. Blood enrichment of TSD. Abdominal aortic blood from six groups of rats was usedto examine Hemoglobin content and number of red blood cells.3. Molecular mechanisms of blood circulation promotion. Five groups were designed,including control, model, drug (TSD9.0g/kg), inhibitor （fasudil10mg/kg） and TSD with inhibitor. LPA concentration was detected to evaluate the status of blood stasis.Related proteins in Rho/Rocks signaling pathway were detected to explore themolecular mechanisms of TSD on blood stasis.Results1. Effective treatment was found for TSD on rat models with BSSPD. TSD improvedthe number of RBC and the content of hemoglobin significantly, reduced the rate ofplatelet aggregation. The concentration of plasma tPA of rat models was increased whilethe plasma PAI-1levels decreased significantly when treated by TSD. The uterine tissuebiopsy of rat models showed an improvement change after treated by TSD, and a higheruterine microvessel density indicated by CD34positive expression also was observedusing the method of immunohistochemistry.2. Molecular mechanism. The concentration of plasma LPA of rat models which is amarker of thrombosis was decreased when treated by TSD, so to further inhibite theLPA/Rho/Rocks/PAI-1signaling pathway, and then made an abnormal expression ofLPA/Rho/Rocks/PAI-1in this pathway. Moreover, the combination of TSD and fasudilcould make a better inhibitory.ConclusionTSD showed an effective treatment on postpartum stasis model rats，and its effect maybe related to the signaling pathways of Rho/Rocks.