The Effect Of PolyCHb Resuscitation On Hematopoietic Function Of Bone Marrow.by Severe Hemorrhagic Shock In Rats

Background:The hypoperfusion after hemorrhagic shock can cause multi-organ dysfunction syndrome,which may lead to abnormal stem cell function in the bone marrow.It is very important to restore the circulating volume,improve ischemia and hypoxia,and recover the oxygen supply balance of the organism as soon as possible.There is not suitable oxygenated solution and explicit guideline pointing out which oxygenated solution is better suited for reviving patients with severe hemorrhagic shock in China at presently.In the preceding work of our project group,we have developed plasma substitute to revive rat with severe hemorrhagic shock which consists of the hydroxyethyl starch and the polyhemoglobin in appropriate proportions.The results obtained are encouraging.However,as hydroxyethyl starch with renal toxicity,many European and American countries do not recommend using it in clinic.Therefore,in this subject,we use human blood albumin mix more portable oxygen poly hemoglobin to treat rats with severe hemorrhagic shock and makes a preliminary discussion on its effects on hematopoietic function.Objective:to investigate the effect of new oxygenated solution which revives rats with severe hemorrhagic shock on bone marrow hematopoietic system,using hemorrhagic shock rats-recovery model.Methods:64 adult male SD rats were randomly divided into 4 groups,including one control group and three interventional groups.(1)control:only to rats move/vein intubation;(2)PolyCHb resuscitation group:rat femoral/venous cannulation,preparation of hemorrhagic shock model,recovery of PolyCHb recovery fluid;(3)erythrocyte recovery group:resuscitated with red blood cell resuscitation,and the remaining operation was the same as PolyCHb group;(4)albumin recovery group:recovery with albumin recovery fluid,the remaining operation is the same as PolyCHb group.The blood pressure,heart rate and counting blood cells were measured at three time points:before bloodletting(baseline value),shock(60 percent of the bleeding was completed immediately),after recovery fluid transfusion.2 h,4 h,6 h,8 h,8 h,12 h,24 h,48 h and 72 h after reviving,0.3ml peripheral blood from rats were collected.Then,peripheral blood was centrifuged and collected plasma immediately.Level of EPO in the separated plasma was measured.24 h and 72 h after the resuscitation,32 rats were killed.The kidney tissue was removed for examination of EPO and EPOR mRNA expression level by qPCR in 24h and 72h.The lung tissue was removed for examination of GM-CSF mRNA expression level by qPCR in 72h.the change of bone marrow cell cycle was measured in72h.The number of erythrocytes in peripheral blood was detected by the test tube method after the anticoagulation of EDTAK2 in 72 h peripheral blood of rats.The sternum of 72 h rats and 4%were fixed.HE staining observed the pathological changes.Results:the experimental group,as a result of severe blood loss shock rats blood pressure,heart rate,PaC02 dropped substantially,Pa02 rise,after fluid resuscitation of each experimental group blood pressure rose to more than 80 MMHG,and heart rate,PaC02 and Pa02 are immediately returned to normal,but Lac and BE not immediately returned to normal,but after 72 h Lac and BE recoverable.With PolyCHb recovery liquid recovery uncontrolled hemorrhagic shock rats can maintain within 6 h after recovery of oxygen to the body to produce more effective,show the EPO slow growth in the albumin group(P<0.05),72 h after recovery kidney EPO mRNA expression is lower than albumin group but higher than that of red blood cells,PolyCHb recovery after 72 h rats reticulocyte number increased significantly in the control group,and no obvious difference with the albumin group,gm-csf PolyCHb recovery after bone marrow cells and IL-3 mRNA expression appeared abnormal rate is higher,However,it was not observed that PolyCHb affected the activity of bone marrow hematopoietic function.Conclusion:PolyCHb can effectively resuscitate rats with severe hemorrhagic shock,and 72 h did not die after resuscitation.PolyCHb used for resuscitation can improve tissue hypoxia and reduce EPO expression in the body,but it did not observe the inhibitory effect on the red hematopoietic system and the abnormal function of bone marrow cells.