| Purpose:This paper is based on the traditional Chinese medicine theory of spleen governs blood,The classic prescription of replenishing qi and blood,invigorating spleen and nourishing heart is selected,The pathogenesis of cellular immunity in Idiopathic Thrombocytopenic Purpura was studied,To observe the effects of Guipi Decoction on the immune balance of Treg cells and Th17 cells,To explore the effect and mechanism of Guipi Decoction in the treatment of ITP,and to provide experimental basis for the clinical treatment of ITP.Material and method:There were 68 BALB/c mice and 10 guinea pigs of SPF grade.After 7 days of adaptive feeding,20 BALB/c mice were used for the preparation of platelet antigens,The other 48 BALB/c mice were divided into 6 groups by random number table method,They were blank control group,model control group,Guipi Decoction low-dose group,Guipi Decoction medium-dose group,Guipi Decoction high-dose group,and western medicine control group.The model was established by intraperitoneal injection of Guinea pig anti-mice platelet serum,A chronic persistent mice model of ITP was established.On the 5th day of modeling,ITP mice were given medication intervention.Among them,Guipi Decoction low-dose,medium-dose and high-dose groups(8.44g·kg-1,16.88g·kg-1,33.76g·kg-1)were given to mice by gavage at different concentrations of Guipi Decoction,respectively;Western medicine control group was given prednisone acetate suspension;Blank control group and model control group were given distilled water by gavage.The medication was administered continuously for 10 days.2 hours after the last intragastric administration,blood was collected by the orbital vein method.In the aseptic state,the spleen was taken and the splenic single cell suspension was prepared by abrasive method;The effect of Guipi Decoction on platelet count in peripheral blood of ITP mice was detected by automatic multi-species five-classification animal haematology analyzer;Flow cytometry was used to detect the effect of Guipi Decoction on the number of Treg cells in peripheral blood of ITP mice;Flow cytometry was used to detect the effect of Guipi Decoction on the number of spleen Treg cells in ITP mice;Flow cytometry was used to detect the effect of Guipi Decoction on the number of Th17 cells in the spleen of ITP mice;Enzyme linked immunosorbent assay was used to detect the effect of Guipi Decoction on the content of TGF-?1 in serum of ITP mice.Results:1 Establishment of ITP mice modelChronic ITP mice model was established by GP-APS method,The peripheral blood platelet count of ITP model mice decreased,and petechiae and ecchymosis appeared on the skin surface,The ITP animal immune model was established by this modeling method for experimental study.2 Effect of Guipi Decoction on peripheral blood platelet count in ITP miceCompared with the blank control group,the peripheral platelet count of the model control group was significantly lower(P<0.01);Compared with the model control group,the peripheral blood platelet count of Guipi Decoction low-dose,medium-dose and high-dose groups and western medicine control group were all increased(P<0.01),Among them,the high dose group of Guipi Decoction had significant curative effect(P<0.01).3 Effect of Guipi Decoction on the number of Treg cells in peripheral blood of ITP miceCompared with blank control group,the number of Treg cells in model control group was decreased(P<0.01);Compared with the model control group,the number of Treg cells in the low-dose,medium-dose and high-dose groups of Guipi Decoction and the western medicine control group were increased(P<0.01),Among them,the high dose group of Guipi Decoction had better effect.4 Effect of Guipi Decoction on the number of spleen Treg cells in ITP miceCompared with blank control group,the number of spleen Treg cells in model control group decreased(P<0.01);Compared with the model control group,the number of spleen Treg cells in the low-dose,medium-dose and high-dose Guipi Decoction groups and the western medicine control group were increased(P<0.01),and the effect in the high-dose Guipi Decoction group was better.5 Effect of Guipi Decoction on the number of Th17 cells in spleen of ITP miceCompared with blank control group,the number of spleen Th17 cells in model control group was increased(P<0.01);Compared with the model control group,the number of Th17cells in spleen of Guipi Decoction low-dose,medium-dose and high-dose groups decreased(P<0.01),Among them,the number of Th17 cells in spleen of Guipi Decoction high-dose group was significantly decreased(P<0.01).6 Effect of Guipi Decoction on serum TGF-?1 content in ITP miceCompared with blank control group,the content of TGF-?1 in model control group was decreased(P<0.01);Compared with model control group,the content of TGF-?1 in low-dose,medium-dose and high-dose Guipi Decoction groups and western medicine control group was significantly increased(P<0.01),Among them,the high dose group of Guipi Decoction had better effect.Conclusion:1 The ITP mice model was successfully replicated by GP-APS injection in vivo.2 The platelet count in peripheral blood of ITP mice decreased,and Guipi Decoction can effectively increase the platelet count in peripheral blood of ITP mice.By comparison,the high dose group of Guipi Decoction had the best effect,it showed a dose-effect dependence.3 Guipi Decoction may down-regulate the number of Th17 cells by increasing the number of Treg cells,increase the content of TGF-?1,inhibit the activation of T cells,and make Treg/Th17 cells approach the immune balance state,Thus effective prevention and control of ITP. |
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