| Immune thrombocytopenia(ITP)is a clinically common hemorrhagic disease.Because the autoimmune system recognizes platelet surface antigen,the number of platelets is decreased.The clinical manifestations are skin purpura and ecchymosis,oral bleeding,and epistaxis.In severe cases,intracranial hemorrhage or visceral hemorrhage may occur,thereby endangering life.In the past,the research on ITP was mainly based on immunity,and there were few studies on blood vessels.Now,vasoactive substances and inflammatory factors are used as the entry point to explore the vascular injury of ITP and ITP with syndrome of QI failing to control blood.At the same time,the clinical prescription Yiqishexue Granules was used for drug administration,and the drug efficacy was observed.Objective:This study is aimed to explore the vascular endothelial injury during the pathogenesis of ITP and ITP with syndrome of QI failing to control blood,and to explore the changes of bleeding mechanism in the pathogenesis of ITP and ITP with syndrome of QI failing to control blood by investigating the changes of vascular active substances and inflammatory factors.The other purpose is to explore the effect of Yiqishexue Granules on vasoactive substances and inflammatory factors in ITP and ITP with syndrome of QI failing to control blood mouse model,and to explore its role in vascular protection.Method:1.ITP mouse model was established by immunization.Balb/C mice were divided into normal group and model group,each group was divided into 3 days and 14 days.The model group was injected with APS every other day,and the normal group was injected with normal saline.At the end of the experiment,the mouse signs were observed and the body weight,grip,platelet count,spleen index,thymus index,vasoactive substances vWF,NO,ET-1,VEGF-A,sICAM-1,sVCAM-1,inflammation were measured.Factor TNF-?,IFN-?,IL-6,IL-4,HE staining of spleen,liver and abdominal aorta.2.A mouse model of ITP with syndrome of QI failing to control blood was established by sleep deprivation combined immunization.Balb/C mice were divided into normal group and model group,each group was divided into 7 days,10 days and 21 days.In the model group,only sleep deprivation was performed in the first week,and APS was injected in the second week,once every other day.Normal group was injected with normal saline.At the end of the experiment,the mouse signs were observed and the body weight,grip,platelet count,spleen index,thymus index,vasoactive substances vWF,NO,ET-1,VEGF-A,sICAM-1,sVCAM-1,inflammation were measured.Factor TNF-?,IFN-?,IL-6,IL-4,HE staining of spleen,liver and abdominal aorta.3.Based on the model of ITP mice,we conduct an efficacy test of Yiqishexue Granules.Balb/C mice were divided into normal group,model group,high-dose group of Yiqishexue Granules(raw medicine 15.22g/kg),low-dose group of Yiqishexue Granules(raw medicine 7.61g/kg),and prednisone group.Drugs were administered the next day after establishing the model.After the end of modeling,the mice were examined for body weight,grip,platelet count,spleen index,vasoactive substances vWF,NO,ET-1,sICAM-1,inflammatory factors IFN-?,IL-6,IL-4,spleen,liver.,abdominal aorta HE staining.4.Based on the model of ITP with syndrome of QI failing to control blood mice,we conduct an efficacy test of Yiqishexue Granules.Balb/C mice were divided into normal group,model group,high-dose group of Yiqishexue Granules(raw medicine 15.22g/kg),low-dose group of Yiqishexue Granules(raw medicine 7.61g/kg),and prednisone group.Drugs were administered the next day after injecting APS.The mice were examined for body weight,grip,platelet count,spleen index,vasoactive substances vWF,NO,ET-1,sICAM-1,inflammatory factor IL-4,HE staining of spleen,liver and abdominal aorta.Result:1.In the ITP model group,the mental state of the mice on the 3rd day was no different from that of the normal group.The skin showed significant bleeding points,and the stool was dark brown with blood in the stool.The grip strength and platelet count were significantly lower than the normal group(P<0.01),the spleen index was significantly higher than the normal group(P<0.01),and the vasoactive substances vWF,NO and ET-1 were significantly higher than the normal group(P<0.01),and sVCAM-1 and VEGF-A were significantly lower than the normal group(sVCAM-1 P<0.05,VEGF-A P<0.01).On the 14th day,the mental state of the model group mice was slightly wilting,the skin had only a small amount of bleeding points,and the stool was dark brown with no blood in the stool.Body weight,grip strength,and platelet count were significantly lower than those in the normal group(P<0.01).The spleen index and thymus index were significantly higher than those in the normal group(P<0.01),and the vasoactive substances vWF,NO,ET-1,and sICAM-1 were higher than those in the normal group(P<0.01),the inflammatory factors IFN-y,IL-6 and IL-4 were higher than the normal group.The results of HE staining showed that the model group had abnormal spleen,red blood marrow expansion and white pulp atrophy,and the germinal center formed.On the 14th day,"starry sky"phenomenon appeared.Inflammatory cell infiltration occurred in the liver of the model group,and focal necrosis occurred with the increase of modeling time.There was a slight lymphocytic infiltration in the abdominal aorta of the model group.2.The mental state of the mice in the model group of ITP with syndrome of QI failing to control blood was sluggish,and their reactions were slow.After stopping deprivation every day,the mice were tired,curled up,rarely moved,their hair was dark yellow and rough,their stools were dark brown or yellow,and their stools were sparse.With the prolongation of deprivation time,they became more and more serious.Body weight,grip strength,platelet count and thymus index were significantly lower than the normal group at three time points(P<0.01).On the 7th day,the spleen index of the model group was significantly decreased(P<0.01),the vasoactive substances v WF,NO,ET-1 were significantly higher than the normal group(P<0.01),On the 10th day,the spleen index of the model group was significantly increased(P<0.01),the vasoactive substances vWF,NO and ET-1 were significantly higher than the normal group(P<0.01),and VEGF-A and sVCAM-1 were significantly lower than the normal group(P<0.01).The inflammatory factor IFN-? was significantly increased(P<0.01).On the 21th day,the spleen index of the model group was significantly increased(P<0.01).The vasoactive substances vWF,NO,ET-1,sICAM-1 were significantly higher than the normal group(P<0.01),and the inflammatory factor IL-4 was significantly increased(P<0.05).The results of HE staining showed that the spleen red pulp was congested in the model group,the white pulp was atrophied,and the germinal center was formed.The phenomenon of "starry sky" after phagocytosis appeared,and the myeloid metaplasia was observed,which increased with the time of modeling.There was lymphocyte infiltration in the model group near the hepatic vein.As the model time increased,hepatic turbidity and necrosis occurred.3.The high-dose group of Yiqishexue Granules had significant effects on vasoactive substances and inflammatory factors in mice.The vasoactive substances vWF,NO,ET-1 and sICAM-1 were significantly lower than those in the model group(P<0.01).inflammatory factors IFN-y,IL-6 and IL-4 were significantly lower than in the model group(P<0.01).The ratio of vWF to ET-1 in the vasoactive substances in the low-dose group of Yiqishexue Granules group was significantly lower than that in the model group(P<0.01),and the ratio of IL-6 and IL-4 in the inflammatory factors was significantly lower than that in the model group.P<0.01).HE staining results showed that high-dose group of Yiqishexue Granules can reduce the infiltration of inflammatory cells such as hepatic venous lymphocytes.4.Yiqishexue Granules can enhance the mental state of mice and effectively increase the platelet and grasping power of mice with ITP with syndrome of QI failing to control blood(P<0.01),and can significantly reduce vasoactive substances such as vWF,NO,ET-1,sICAM-1(P<0.01),significantly reduced the inflammatory factor IL-4(P<0.01).HE staining results showed that high-dose group of Yiqishexue Granules can reduce the infiltration of inflammatory cells such as hepatic venous lymphocytes.Conclusion:1.ITP model mice showed a decline in physical function,but in good mental state.It is believed that some symptoms of Qi deficiency have appeared.Injection of APS resulted in endothelial injury and abnormal vasoactive substances,which aggravated the abnormalities of immune system and inflammatory reaction with the increase of modeling time.2.During the pathogenesis of ITP with syndrome of QI failing to control blood model mice,obvious Qi-de:ficiency and bleeding appeared,which indicated that the model was effective.The endothelial vessels of mice have been damaged to a certain extent and the endothelial environment has been damaged after only sleep deprivation.Injection of APS and continuous deprivation maintain the sustained damage.On the 10th day(the 3rd day after APS injection),endothelial cells were damaged and bleeding was the most serious because of the rapid decrease of platelets.On the 21 st day,the inflammatory reaction of mice was enhanced.3.Yiqi Shenxue Granule can treat ITP and ITP Qi-deficiency syndrome by regulating vasoactive substances and inflammatory factors. |
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