| Objective The changes of oral mucosa histopathology,body weight,clinical manifestations and the changes after high,medium and low dose Yukouning were observed.The pathogenesis of rat model was studied at molecular level.Yukouning was used to regulate T The mechanism of h1/th2 immune balance and the correlation with JAK/STAT signaling pathway provide experimental basis for Yukouning in the prevention and treatment of cardiosplenic hot RAU.Method1.theoretical research:Through consulting the relevant literature at home and abroad,this paper discusses the name of Chinese medicine,pathogenesis,syndrome differentiation and treatment of RAU,and discusses the origin,etiology,clinical manifestations and treatment methods of RAU from the perspective of Western medicine.2.experimental study:After one week adaptive feeding,60 SD rats were randomly anesthetized with 10% chloraldehyde hydrate,and the cervical dislocation was killed to prepare the oral mucosa tissue homogenate.The immune antigen emulsifier was obtained by mixing the oral mucosa homogenate with Freund complete adjuvant at 1:1 ratio.The remaining 50 rats were randomly divided into 5 groups,control group,model group,Yukouning high dose group,Yukouning medium dose group and Yukouning low dose group,each group 10 rats.Except for the control group(single cage granule feed for free drinking water),the single cage high fat spicy feed was used to feed the feed and wear neck ring.After 14 days,the back hair of the four groups was hair removed,the immune antigen emulsifier was injected into the skin,and the antigen was injected once a week for 8 weeks.On the54 th day after the first injection,the lower lip mucosa was burned with 50% glacial acetic acid for 30 s,and 48 hours later,the rats were selected.The main symptoms and signs of rats,histopathological changes,T-lymphocyte changes in spleen of each group were detected by flow cytometry,real-time fluorescence quantitative PCR and Western blot analysis were performed in oral mucosa.SPSS 23.0 statistical software was used to analyze the data.Result1.main symptoms and signs of rats:After the model of the syndrome of cardiosplenic heat,the rats had rough body hair,irritable temperament and some noses.However,the blood mark of nostril was seen,the gingiva was slightly red and swollen,the teeth were dry,the stool was dry,the stool was yellow,the reaction was slow,the amount of food and drinking water decreased,and the lower lip mucosa appeared round or oval ulcer.Compared with the control group,the weight of the model group decreased slowly after stopping the stimulation.The weight of the low dose Yukouning group did not increase significantly,showing a trend of fluctuation,while the weight of the high and medium dose groups was on the rise.2.HE′s changes:The results showed that the epithelium,basement membrane and propria layer of oral mucosa in the control group were clear and the structure was obvious.The repair of epithelial mucosa was observed in each group compared with the model group.The edema in epithelial cells and between cells was observed in the low dose group and model group,and the basement membrane was clear.3.the changes of T-lymphocyte in spleen of rats were detected by flow cytometryCompared with the control group,the spleen of model group showed that the production rate of IFN-γ T cells(Th1)increased,while after the treatment of drugs,the cell generation rate decreased significantly.In contrast,the model group showed a completely different trend in Th2 cells,and the production rate of IL-4 T cells(Th2)was significantly decreased compared with the control group.The down-regulation rate of Th2 cells was on the rise after the treatment.The results suggest that the high,medium and low dose of Yukouning can regulate the balance of Th1/Th2 in the spleen of the rat with cardiosplenic heat by increasing the expression of Th2 in spleen and inhibiting the expression of Th1 in spleen.4.real time fluorescence quantitative PCR detection of oral mucosa Compared with the control group,IFN-γ expression in the rat model of cardiosplenic heat type RAU increased.However,after Yukouning was given to each group,the expression of IFN-γ m RNA decreased significantly.In the oral mucosa of RAU with cardiosplenic fever,T-bet showed a decrease in m RNA expression after drug treatment,which promoted the expression of GATA-3 m RNA.These results showed that Yukouning could prevent inflammation of RAU oral mucosa of cardiosplenic fever by increasing the m RNA expression of th2-related cytokines and transcription factors.5.Western blot analysis of oral mucosaIn the model group,the expression of p-stat1 was up regulated,and they were significantly down regulated after administration.However,the total expression of STAT1 in oral mucosa of rats with cardiosplenic heat type RAU was not changed.Compared with the control group,the expression of p-stat6 was down regulated in the model group.The decrease was significantly increased after Yukouning was given to each group,but this did not lead to the change of total STAT6 expression.In conclusion,these results showed that the changes of JAK/ STAT signal pathway could clarify the effect of biological activity of Yukouning on th1/th2 immune response in rats with cardiosplenic heat.Conclusion1.the most common syndrome type of recurrent apherta ulcer is the type of accumulation of heat in the heart and spleen.It is advisable to clear the heat and detoxify the toxin,eliminate swelling and pain,and the application of yukoning is effective.2.The clinical symptoms of recurrent apherta ulcer could be found by immunochemistry combined with high fat and spicy feed and wearing neck ring.The syndrome was cardiosplenic heat accumulation type,suggesting that the model could be copied successfully.3.In RAU oral mucosa,it was found that Th1 cells were concentrated in large quantities through activating IFN-γ/STAT1 pathway by p-stat1;Th2 cells were relatively deficient,but IL-4 depended on STAT6 to induce Th2 cell differentiation,and then activated transcription factor GATA-3,suggesting that the pathogenesis of RAU might be achieved by JAK/STAT signaling pathway.4.Yukouning inhibited IFN-γ/STAT1 pathway by inhibiting STAT1 phosphorylation,activated p-stat6 to regulate Th1/Th2 balance.It is suggested that the mechanism of Yukouning,a traditional Chinese medicine prescription,to improve the RAU rats with cardiosplenic heat accumulation may be related to the regulation of JAK/STAT signaling pathway. |
