Expression Of HMGB1 In MODS Rats Treated With BMSC

Part One The Model Establishment of MODS in SD ratsObjective Campare different methods of the multiple organ dysfunction syndrome rats models in order to define a most appropriate and stable MODS rats model for the later research.Methods Healthy adult female SD rats were randomly divided into 9 groups, each group 20, which weighted between 190g and 220g. Group A:Mild anemia (lost 15% body blood volume)+given lipopolysaccharide intraperitoneally30mg/kg. Group B: Mild anemia (lost 15% body blood volume)+given lipopolysaccharide intravenously 5mg/kg; Group C:given lipopolysaccharideintravenously 5mg/kg; Group D:given lipopolysaccharide intravenously 4mg/kg; Group E:given lipopolysaccharide intravenously 3mg/kg; Group F:given lipopolysaccharide intravenously 2mg/kg; Group G:given lipopolysaccharide intravenously lmg/kg; Group1 H:given lipopolysaccharide intravenously (0.5mg/kg);Sham-operated group I were injected PBS0.5ml/200g intravenously. Basic presence and mortality rate were recorded after modeling, hepatic function(ALT、AST)、renal function(BUN、CR)、myocardial enzyme(CK-MB、LDH) and routine blood tests(WBC、PLT) were also recorded at 24h by blood drawing, then dissected the rats and compared with pathological section, finally get the optimal policy.Results Compration among groups, Mortality rate in group A B and C are the same, were all 80%, group F and G were both 60%, group H was 40%,group I was 0; the results of hepatic function、renal function、myocardial enzyme and routine blood tests in group A-G were obvious different when compared with group I, while there was no difference when compare group H with group I; body samples displayed that organs in group A-C damaged seriously with congestion、edema and congestion; pathological sections showed that there was acute injury performance like inflammatory cells infiltration and hemorrhage or edema in group A-GConclusion Animals in group F and G had a higher survival rate, and the animals experienced multiple organ dysfunction, while group G using lower dosage of lipopolysaccharide, thus, the method of group G can be an optimal policy for making MODS models.Part Two Expression of HMGB1 in MODS rats treated with BMSCObjective:Study the expression levels and localization changes of high mobility group protein B1 (HMGB1) and other inflammatory cytokines in multiple organ dysfunction syndrome (MODS) rats by bone mesenchymal stem cells (BMSCs) transplantation to explore the possible mechanism of the BMSC transplantation in the treatment of MODS rats.Methods:BMSC was obtained and cultured in vitro, labeled with green fluorescent protein (GFP),transplanted by intravenous infusion.Adult female SD rats were randomly divided into 3 groups,10 rats in the normal group,40 rats in model group, transplantation group 40.2 hours after LPS(lmg/kg) intravenous injection molding, stem cells (2*10∧6/ml*0.5ml)were transplanted by intravenous infusion. On the first day, for transplantation group and model group, the heart, liver, spleen, lung, kidney and intestinal tissue sections were taken to observe GFP-BMSC homing in MODS rats and HMGB1 distribution in MODS rats respectively; On the 1,3,7,14 days, heart, liver, spleen, lung, kidney and intestinal tissue were taken to test the HMGB1 expression changes through Q-PCR and Western blotting technique.Results:In vitro cultured BMSC have a typical mesenchymal stem cells pattern, also have the ability to differentiate into osteoblasts and adipogenic cells. It was not changed that the differentiation potency after lentivirus transfection.BMSC was CD34-/CD45-/CD90+/CD44+that conformed to the typical characteristics of BMSC. Green fluorescence was found in the heart, liver, spleen, lung and kidney of recipient on first day, proved that BMSC homing has occueeed; Immunohistochemical results showed that HMGB1 in the model group transfers to cytoplasmic; through Q-PCR and Westernblot testing, to normal group, the HMGBl expression level in heart, liver, spleen, lung, kidney tissues of model group was mostly significantly increased (P<0.05), while the treatment group mostly lower than model group but higher than normal group (P<0.05).Conclusion(s):①Established in vitro culture system and GFP-mark technology of rat BMSC. ②BMSC transformation could reduced the expression of HMGB1 in the MODS heart, liver, spleen, lung, kidney tissues, which remind BMSC transplantation can inhibit inflammatory reactionary may be on new method in treatment of MODS.