| ObjectiveAccording to the theory of Spleen governing blood,analyzed articles to expound and elaborate the connotation of this theory in hemorrhagic diseases,and expand the theory to further guide basic research and clinical application;Prepared ITP animal model by injected GP-APS,to explore the effect of JianPi YiQi SheXue formula on ITP and its immunity network regulation mechanism.MethodsAnalyze the discourse of spleen governing blood in ancient books of TCM and modern research,discuss its role of guiding in basic research and clinical application.Guinea pig anti-mouse platelet antiserum(GP-APS)was prepared.Pre-process the serum and test the antibody titers.BALB/c mice were randomly divided into two groups by blood platelet count.By intraperitoneal injection of GP-APS,blood from caudal was collected vein before injection and 6h,12h,48h after the injection to determine whether the model is successful or not.Then 80 mice were divided into 4 groups:normal group,model group,prednisone group and Jianpi Yiqi group.The GP-APS was given by intraperitoneal injection to BALB/c mice in experimental groups every day to establish ITP animal model.And normal saline was given to the mice in normal group.Prednisone and Yiqi Jianpi Shexue formula was given by intragastric administration to the mice every day from the 8th day?General behavior,morphological changes,peripheral blood cell count,thymus and spleen index of the mice were observed to evaluate the therapeutic effects of Yiqi Jianpi Shexue formula.The content of TGF-?1,IFN-?,IL-4,IgA,?-EP,5-HT,VIP and NE in serum,intestinal tissues or brain tissues were determined by ELISA assay or IHC assay.ResultsIn a narrow sense,it means keeping the blood flowing in the vessels.In the broad sense,it should be understand from four aspects:engendering blood,moving blood,arresting bleeding,and wrapping blood.All of these aspects can explain the function of the spleen in physiology and pathology.Curative effect evaluation of the Yiqi Jianpi Shexue Formula experimental intervention to ITP model mice as follows:There was no remarkable difference in swimming time,temperature,weight and intake among three groups(p>0.05).The platelet count of Jianpi Yiqi group and prednisone group were higher than that in model group(p<0.05),There was no remarkable difference in HGB,WBC count and PDW among three groups(p>0.05).The thymus index of model group and Jianpi Yiqi group were higher than that in model group(p<0.05),and there was no remarkable difference between prednisone group and normal group.The spleen index of model group,prednisone group and Jianpi Yiqi group was higher than normal group(p<0.05),and also prednisone group was lower than model group(p<0.05).The effect of YiQi JianPi SheXue formula on ITP model mice by immunity network regulation mechanism as follows:There was no remarkable difference in TGF-?1 concentration among the groups(p>0.05).IFN-? concentration of model group and prednisone group was higher than that in normal group,Jianpi Yiqi group was lower than normal group and model group,differences were statistical significance(p<0.05).IL-4 concentration of model group and Chinese group was lower than normal group,and prednisone group was higher than model group,differences were statistical significance(p<0.05).The IFN-?/IL-4 ratio of model group was significantly higher than normal group(p<0.05).The ratio of prednisone group and Chinese group was lower than model group(p<0.05),the difference between prednisone group,Chinese group and normal group,was not statistically significant.The serum concentration of IgA in model group and Jianpi Yiqi group was higher than that in normal group,it was lower in prednisone group than model group,the differences were statistically significant(p<0.05),The IgA content in small intestine in model group and prednisone group was lower than that in normal group,and that in Chinese group was higher than model group,the differences were statistically significant(p<0.05).The serum concentration of ?-EP in model group was higher than that in normal group,it in Jianpi Yiqi group was lower than in model group,the differences were statistically significant(p<0.05).The serum concentration of 5-HT in model group was lower than that in normal group,it in prednisone group and Jianpi Yiqi group was higher than in model group,the differences were statistically significant(p<0.05).The serum concentration of VIP in model group was lower than that in normal group,it in prednisone group and Jianpi Yiqi group was higher than in model group,the differences were statistically significant(p<0.05).The serum concentration of NE in model group,prednisone group and Jianpi Yiqi group was higher than in normal group,the differences were statistically significant(p<0.05).The concentration in intestinal tissue of ?-EP in model group was lower than that in normal group,it in Jianpi Yiqi group was higher than in model group,the differences were statistically significant(p<0.05).The concentration in intestinal tissue of 5-HT in model group was lower than that in normal group,it in Jianpi Yiqi group was higher than in normal group,it in prednisone group and Jianpi Yiqi group was higher than model group,the differences were statistically significant(p<0.05).The concentration in intestinal tissue of VIP in model group was lower than that in normal group,it in Jianpi Yiqi group was higher than in model group,the differences were statistically significant(p<0.05).The concentration in intestinal tissue of NE in Jianpi Yiqi group was higher than that in model group,the differences were statistically significant(p<0.05).There was no remarkable difference in?-EP concentration in brain tissue among the groups(p>0.05).There was no remarkable difference in 5-HT concentration in brain tissue among the groups(p>0.05).The concentration in brain tissue of VIP in Jianpi Yiqi group was lower than that in model group,the differences were statistically significant(p<0.05).The concentration in brain tissue of NE in prednisone group was higher than that in model group,the differences were statistically significant(p<0.05).ConclusionThe theory of Spleen governing blood should also contain engendering blood,moving blood,arresting bleeding,and wrapping blood.According to this theory,"Strengthening Spleen Replenishing Qi and Control blood" is the main therapeutic principle of chronic ITP.Jianpi Yiqi Shexue formula can efficiently raise the platelet count in ITP mouse model as prednisone,and regulating immunity network of ITP model is the mechanism of Jianpi Yiqi Shexue formula. |
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