The Effects Of Bone Marrow Mesenchymal Stem Cells In Cerebral Resuscitation And Expressions Of NGF、Caspase-3 After Cardiac Arrest In Rats

Cardiac arrest is the world’s leading cause of death in emergency department,and there are many diseases which can lead to cardiac arrest. With modern technology of cardiopulmonary resuscitation become more and more standardize and emergency network constantly improve,circulation recovery rate after cardiopulmonary resuscitation has been greatly improved。As the brain cells is particularly vulnerable under the condition of hypoxia,so many patients are still in a coma after spontaneous circulation recovery, so cerebral resuscitation has become the most important factor which influences the survival rate of cardiopulmonary resuscitation patients after cardiac arrest.Therefore, to explore the effective cerebral resuscitation after cardiac arrest become an important and meaningful area in Emergency MedicineBone Marrow mesenchymal stem cells(BMSCs) are stromal cells which have multi-differentiation potential and are distinct from the hematopoietic lineage.Many Studies show that BMSCs can save the neurons, induce MSC-mediated neural functional recovery through secreting trophic factor and reducing apotosis.At present, only a few researches have done. However, all of them have indicated that MSCs transplantation significantly improve neurological outcome.Objective of the experiment:To explore the effects of BMSCs on the expression of NGF、Caspase-3 in rat followed Cardiac arrest and the potential mechanisms that BMSCs effect on Cerebral Resuscitation.Methed and Result of the experiment:1. BMSCs’ culture and identification.Choosed the SD rats weight from 100 g to 110 g,executed though cervical,the marrow cavity with DMEM/F12 culture solution, and then collected the bone marrow.BMSCs which were isolated from rat bone marrow and purified by means of attachment culture method.The microscope was applied to observe the cellular morphology in Passage 3.The cells had a similar size, spindled shape,fitted closely and whirlpool growth after 8-9 days. The 3rd generation cells were tested by flow cytometry. The expressions of CD29, CD90 were 92.38%, 99.21%. The expressions of CD45, CD11 b were 0.21%, 0.18%.It is indicated that use the attachment culture method can be used as a reliable method to isolate and purify the BMSCs,2. The potential efficacy of transplanted bone marrow derived mesenchymal stem cells in expression of NGF、caspase-3 and cerebral resuscitation.18 male SD rats were randomly divided into 3 groups(each 6).Group A(Sham group),group B(CA),group C(CA+BMSCs group). One hour after ROSC, group C injected with a dose of 1×106/ml BMSCs via the vena caudalis。Then observed the survival of the rats.The rats in each groups were sacrificed 7 days after cardiac arrest.The neurological status after ROSC was assessed by modified neurological severity scores(m NSS) and serum levels of S100 B were recorded in 7d after ROSC..At the same time,the expressions of NGF and Caspase-3 in brain tissue were assessed by immunohistochemistry to discuss the possible mechanism of the nerve function’s improvement.Through the m NSS test, the m NSS score of cardiac arrest group was higher than sham operation group significantly after ROSC at 1st day and 3rd day(p<0.05), the m NSS score of BMSCs treatment group was reduced significantly(p<0.05). Compared with sham operation group,the m NSS score of BMSCs treatment group was no statistical difference at 7th day(p>0.05). Serum level of S100 B of cell-transplanted group and cardiac arrest group was higher than sham operation group(p<0.05). Cell-transplanted group decreased significantly after ROSC than cardiac arrest group at the content of S100B(p<0.05).The results suggested that BMSCs transplantation may improve neural functional in rats after cardiac arrest and BMSCs transplantation increased the expression of NGF,and reduced the level of Caspase-3.Conclusion:1. The conditions of isolating,cultivating purifying and proliferating BMSCs were optimized in present study.Though this method can get high purity and activity BMSCs which conformd to international standard.2. BMSCs transplantation increased the expression of NGF, and reduced the level of Caspase-3 to promote the nerval function’srecovery after Cardiac arrest.