Treatment Of The Inflammation After Aneurismal Subarachnoid Hemorrhage: Transunion Of Regulatory T-cells

Objective1、To make a rat model constructed of subarachnoid hemorrhage that is accepted theCD4+CD25+regulatory T cells transplantation therapy;To research the immunomodulatorycerebroprotective effects of regulatory T cells transplantation in subarachnoid hemorrhagerats by observing the inflammatory cell infiltration.2、To research the influence of regulatory T cells transplantation on inflammatorycytokines in peripheral and inflammatory cells infiltration in CNS in subarachnoid hemo-rrhage rats.Methods1、Magnetic cell sorting (MACS) two-steps method to extract healthy adult wister rat’sCD4+CD25+regulatory T cells. Methods Healthy Wistar rats were used in the experiment.Autologus arterial hemolysates were injected into rat cisterna magna to induce SAH. Therats were divided into the normal group;the sham-operated control group (sham),SAHgroup was divided into PBS treatment group,SP treatment group and Tregs treatmentgroup.After48hours of SAH,the brain was preparation of paraffin sections.Immunohistochemical staining of neutrophils distribution in the brain tissue.2、Healthy Wistar rats were used in the experiment. Autologus arterial hemolysateswere injected into rat cisterna magna to induce SAH,The rats were divided into the normalgroup,the sham-operated control group(sham),SAH group were divided into PBS treatmentgroup,SP treatment group and Tregs treatment group. After48hours of SAH, Thedistribution and the dynamic expressions of TLR4and NF-κB were observd by opticalmicroscope after immunohistochemistry staining. Analyze the expression of TLR4andNF-κB among the groups by Western blot.Enzyme-linked immunosorbent assay (ELISA) evaluate the expression level of inflammatory cytokines IL-1β、TNF-α、IL-6in plasma.ResultCompared with the normal control group and sham group,TLR4-like immunostainingwas present extensively in the brain after SAH. At the same time,the level of NF-κB andIL-6in SAH group was significantly increased (P0.05),Tregs treatment group comparedwith the PBS treatment group and SP treatment group,the level of TLR4,NF-κB in thebrain tissue and the expression level of inflammatory cytokines IL-1β、TNF-α、IL-6inplasma was significantly lower(P0.05).ConclusionWe have characterized the expression pattern of proinflammatory cytokines and TLR4with a link of NF-κB signaling in the brain injury after experimental SAH. Inflammatoryresponses mediated mainly by the activation of the TLR4/NF-κB signaling pathway mayplay an important role in the pathogenesis of acute brain damage following SAH.Regulatory T-cells transplantation by vein significantly reduce the level of TLR4,NF-κB inthe brain tissue and the level of inflammatory cytokines IL-1β、TNF-α、IL-6in plasma Itsuggested that Regulatory T-cells limiting parenchyma damage in autoimmune centralnervous system inflammation.