Research The Mechanism Of Modified Shoutai Pills On Combination Of Disease And Symptom SA Model About Reducing The Fetus Loss

ObjectiveTo study the mechanism of modified Shoutai pills about reducing the fetus loss, with the modified Shoutai pills high dose, middle dose and low dose (hereinafter referred to as high dose, middle dose and low dose), the positive control drug CsA four drugs to intervent the combination of disease and symptom SA model, by ELISA technique to detect serum, HE staining to observe morpho logical changes, RT-qPCR to detect gene, IHC and WB method to detect protein to research the differences among groups of pregnant mice, so as to explore if drugs intervention reversing the the balance of Th17/Treg bias of this model.MethodsTo observe the general situation during the process of experiment about the weight of pregnant mice, the rate of the fetus loss, to detect the level of serum IL-17 among different groups with ELISA method.To detecte such mRNA as IL-17 mRNA, Foxp3 mRNA, ROR gamma t mRNA, and PR mRNA among different groups with RT-qPCR technology.To detecte such protein as Foxp3^ IL-17> PR^ and ROR gamma t protein with IHC and WB technology.ResultsWeight:Normal, high, medium, low dose and positive group of mice are in good condition. Pregnancy day 0,4.5 day,9 day, the weight among groups of pregnant mice is no difference (P> 0.05), but the differences within the groups are significant (P<0.01).The fetus loss:The difference rate of fetus loss between the first, the second model group and the blank group is significant (P< 0.0016);the second model group, the high dose group and the the first model group is no difference (P> 0.0016);the differernce of the middle, low, positive control group and the first group is statistic (P< 0.0016), but the difference among them is no difference (P> 0.0016).The IL-17 serum:The difference between the first model group, the second model group and the blank group is significant (P< 0.01);the second model group, the high dose group, the medium dose group, the low dose group, the positive and the the first group is statistic(P< 0.01). There is no tendency diffenerce among the drug therapy groups.The mRNA:Comparing the the first model group, the second model group with the blank group:The Foxp3 mRNA and and the PR mRNA are down regulated significantly, but the drug therapy groups are up regulated significantly;and the situation of the IL-17 mRNA and the ROR gamma t mRNA among groups are on the contrary.The IHC results:Comparing the the first model group, the second model group with the blank group:The Foxp3 protein and the PR protein are down regulated signifycantl y, but the drug therapy groups are up regulated significantly;and the situ ation of the IL-17 protein and the ROR gamma t protein among groups are on the contrary. There is no tendency diffenerce among the drug therapy groups about the Foxp3 protein and the IL-17 protein. The high dose, middle dose and low dose groups are better than the the positive control group, but There is no tendency diff enerce among the three Chinese drug therapy groups.The WB results:Comparing the the first model group, the second model group with the blank group:The Foxp3 protein and the PR protein are down regulated significantl y, but the drug therapy groups are up regulated significantly;and the situ ation of the IL-17 protein and the ROR gamma t protein among groups are on the contrary. There is no tendency diffenerce among the drug therapy groups about the PR protein. The high dose, middle dose and low dose groups are better than the the positive control group about the ROR gamma t protein and the IL-17 protein group, and there is tendency diffenerce among the three Chinese drug therapy groups.ConclusionI created the combination of disease and symptom SA model and get the SOP about this model, and the Foxp3 mRNA, PR mRNA, Foxp3 protein and the PR protein are down regulated significantly, but the IL-17mRNA, ROR gamma t mRNA, the IL-17 protein and the ROR gamma t protein are up regulated significantlyThe Foxp3 mRNA, PR mRNA, Foxp3 protein and the PR protein are UP regulated significantly, but the IL-17mRNA, ROR gamma t mRNA, the IL-17 protein and the ROR gamma t protein are down regulated significantly about the high dose, middle dose and low dose groups.According to the rate of the fetus loss, we can deduct that Foxp3^ IL-^ PR、ROR γt maybe the therapeutic target of the model in this experiment.Aout this experient we can know:Th17 secrets IL-17 highly and specifi citily, ROR gamma t is specific molecular transcription of Th17, both of them highly expressing on behalf of the Th17 cytokine highly expressing and its dominant position; Foxp3 is specific molecular transcription of Treg, its high expression of Foxp3 representes Treg high expression of cytokines and its advantages, combining with the experiment result shows that Th17 cytokines advantage expression of the model;After drug intervention, the expression of this advantage reverses, presents the advantage of Treg cells factor expre ssion, so that drug intervention group model Th17 cytokines advantage expre ssion reverses to the advantage of Treg cells factor expression, thus corre cting the Th17/Treg balance deviation of the two model groups.