Polarization Of Mononuclear Macrophages Transplanted Into Rabbit Subarachnoid Hemorrhage Model

ObjectiveThe animal model of subarachnoid hemorrhage(SAH)was established in healthy New Zealand rabbits.The isolated mononuclear macrophages from peripheral blood of rabbits were transplanted into the subarachnoid cavity of the model rabbits.The samples of cerebrospinal fluid were extracted for transcriptome sequencing analysis.The differentially expressed genes were screened and their biological functions were analyzed to study the subarachnoid hemorrhage.The effects of macrophage transplantation on polarization and disease were studied to explore the therapeutic effect of cell therapy on subarachnoid hemorrhage and the possibility of clinical application.Method1.A SAH model was established by injecting autologous arterial blood into the occipital cistern(cerebellomedullary cistern)of healthy New Zealand rabbits.The dietary status,general performance and activities of the rabbits were observed.2.New Zealand rabbits were randomly divided into two groups: experimental group(isolated rabbit peripheral blood mononuclear macrophages were implanted into SAH model rabbit subarachnoid space),control group(saline was injected into SAH model rabbit subarachnoid space),and rabbit cerebrospinal fluid was extracted on 1 day,3days,7 days,14 days for transcription group sequencing and data analysis.Result1.After autologous blood was injected into the model,the mental state of the rabbits became worse and more agitated,and some of them had convulsions.The cerebrospinal fluid obtained after blood injection was bloody,and the cerebrospinal fluid of both groups was clear on the 14 th day.2.All samples were quality-controlled,Q30 was more than 90%,which met the needs of the experiment.The uniform distribution of transcript coverage was correct,and there was no 5’/3 deviation.3.Within one day,three days and seven days,the number of differentially expressed genes increased with time,which was mainly related to immune response,cell metabolism,biosynthesis,cell cycle and gene expression.4.In the experimental group,the M1 polarization factors CD163,TNF-＿,IL-6 and IL-1beta were all up-regulated,while the M2 polarization factor TGFB3 was up-regulated on the 7th day.ConclusionIn the early stage(1-3 days)after SAH,M1 polarization was dominant,and M2 polarization factor was up-regulated at 7 days.