The Effect Of Activation Of Nrf2-ARE Pathway In The AGVHD And Mechanism Study

Objective:Hematopoietic stem cell transplantation is a significant therapy method for malignant hematological patients, but the complication, graft versus host disease(GVHD), obviously limited it’s development and clinical application; in this study, we establish the GVHD model, and further use sulforaphane to activate the Nrf2-ARE signal pathway, according to exploring the relationship between Nrf2-ARE and aGVHD to find a new therapy target which prevent and manage the aGVHD and to bring hope for those patients who accepted hematopoietic stem cell transplantation.Methods:1. We used C57BL/6and BALB/c mice to establish the GVHD modal, and injected bone marrow cells and spleen mononuclear cells which separated from donor mice into the caudal vein of recipient mice which accepted total body irradiation (TBI) before transplantation At the same time, setting the group BM(bone marrow cells transplantation) and the group TBI(total body irradiation). GVHD models is divided into two groups:(1) group GVHD-SFN:sulforaphane10mg/kg i.p Qod,(2) group GVHD-NS:saline0.2ml i.p Qod. Evaluating the clinical scores by recording clinical symptoms, body weight and survival time.2. establishing modal and dividing the groups as the mentioned above. The mice were sacrificed at14days after transplantation, detecting the expression of Nrf2, NQO1, HO-1gene by QPCR and the protein expression of Nrf2, NQO1, HO-1by Western Blot; observing the pathological changes of liver, intestine, lung by HE stain and comparing the difference among group GVHD, group BM; Elisa is used to detect the levels of TNF-a and IFN-y.Results:1.the survival time of group GVHD-SFN is higher than group GVHD-NS, the body weight losing of group GVHD-NS is more than group GVHD-SFN, and group GVHD-NS have higher clinical scores than group GVHD-SFN; the number of inflammatory cells of group GVHD-NS more than other groups, the group GVHD-NS has worse pathological changes than group GVHD-SFN.2.the mRNA and protein levels of Nrf2, NQO1, and HO-1of group GVHD-SFN are higher than group GVHD-NS in small intestine tissues.3. The plasma levels of IFN-γ and TNF-α have no significant difference between group GVHD-NS and group GVHD-SFN.Conclusions:1. Sulforaphane can up-regulate Nrf2gene expression and activate the downstream anti-oxidant gene NQO1and HO-1expression.2. The activated Nrf2-ARE signal pathway can reduce aGVHD of the transplanted mice.3. The activated Nrf2-ARE signal pathway does not affect the levels of IFN-γ and TNF-α in mice plasma.