Regulation of non-classical immune parameters in immune thrombocytopenic purpura mice by a spleen-invigorating,qi-replenishing and blood-containing formula

Objective:This study investigated the regulatory effect of non-classical immune parameters on immune thrombocytopenic purpura(ITP)mice by a spleen-invigorating,qi-replenishing and blood-containing formula(SQBF).Method:A total of 80 BALB/c mice were randomly divided into four equal groups(20 mice each):control group,model group,prednisone group and spleen-invigorating,qi-replenishing and blood-containing(SQBF)group.Mice in the model group,prednisone group,and SQBF group were administered anti-platelet serum to induce ITP.The dynamic variations of platelet counts in ITP mice were measured with an automatic blood analyzer before modeling and 48 h,and 8,12 and 15 days following APS injection.Levels of b-endorphin(b-EP),vasoactive intestinal peptide(VIP)and salivary IgA(SIgA)were detected by enzyme-linked immunosorbent assay(ELISA)on 15th day of experiment.Results:SQBF enhanced peripheral blood platelet counts in ITP mice similar to that of prednisone,and both groups showed a statistically significant response compared with the model group(P<.01).The SQBF significantly decreased b-EP levels compared with the model and prednisone intervention groups(P<.05),significantly increased the levels of VIP and SIgA in ITP mice compared with the model group(P<.05)and had significant protective effects on the thymus of ITP mice compared with the model group(P<.01).Conclusions:The SQBF had a similar effect to prednisone with regards to enhancing peripheral blood platelet counts in ITP mice.Furthermore,it decreased b-EP levels and increased VIP and SIgA,and protected the thymus.This shows that,on base of the brain-gut axis functions,some non-classical immune vascular active factors or neurotransmitters are also involved in immune responses,and also have relationship with the onset of ITP and bleeding and/or hemostasis.It needs further study to determine whether a change in these active factors is related to immediate hemostasis.

Investigation of hemostatic effect of Spleeninvigorating,Qi-replenishing and Bloodarresting Formula on simvastatin-induced zebrafish hemorrhage model

Blood-arresting Formula on simvastatin-induced zebrafish hemorrhage model compared with that of the Heat-clearing and Blood-cooling Formula.Methods:AB strain wild type zebrafish were treated with 0.5 mM simvastatin for 24 hours to establish the zebrafish hemorrhage model.Under strictly blind experimental conditions,the zebrafish were then treated with experimental agents in different concentration within the maximum non-lethal dose.The intervention effect of the Spleen-invigorating,Qi-replenishing and Blood-arresting Formula was comprehensively assessed by examining the main observational parameters such as the bleeding reduction rate and hemostatic rate while referring to the additional parameters such as blood flow,improvement rate of blood flow,velocity of movement,improvement rate of motion under spleen-qi deficiency.Results:Hemostatic effect:The bleeding rate and hemostatic rate in the zebrafish were respectively,30%,15%,45%,40%and 60%,80%,40%,47%when equal concentration of 500 mg/mL,1000 mg/mL were used.This showed that the experimental herbs B1 was superior to B2 in decreasing the bleeding rate and improving hemostatic effect in the zebrafish.Improvement of blood flow:With equal concentration,the experimental herb B1 was superior to B2 in increasing the blood flow and improvement rate of blood flow in hemorrhagic zebrafish.Promotion of the improvement of motion:With equal concentration of 500 mg/mL and 1000 mg/mL,the experimental herb B1 was superior to B2 in promoting the velocity of motion in the zebrafish and promoting the improvement rate of motion.Conclusion:The manifestations of simvastatin-induced zebrafish hemorrhage are basically similar to that of failure of the spleen to control blood in human beings.The Spleeninvigorating,Qi-replenishing and Blood-arresting Formula has good hemostatic effect on simvastatin-induced hemorrhage in the zebrafish.It also boasts the functions of improving blood flow and velocity of motion in the hemorrhagic zebrafish.Such a conclusion provides the experimental basis for the treatment of the syndrome of failure of the spleen to control blood by this formula.