| Background Severe acute pancreatitis(severe acute pancreatitis, SAP) is a common clinical digestive diseases, of which 20%-25% of patients may have systemic complications, multiple organ failure and even life-threatening. SAP treatments including internal medicine, surgery, Chinese medicine, etc, but there is still no effective measures to cure, besides, the disease has a rapid onset and high morbidity, the mechanism is not fully understood, SAP complicated by systemic inflammatory response syndrome. Therefore, look for specific targets for SIRS suppress the development of inflammation could be a way out. Objective1. To explore the relationship between TNFAIP3/A20 protein and IKK/IκB/NF-ΚB signaling pathway early activation status in the pancreas and liver tissues of SAP(severe acute pancreatitis, SAP) rats.2. To discuss IκB/NF-ΚB signaling pathway and the expression of EPRS protein in the pancreas and liver tissue in early onset of SAP-related pathogenesis.Methods1.Retrograde cholangiopancreatography injection of 5% sodium taurocholate established SAP model(1) 96 adult SD(Sprague-Dawley) rats, female were randomly divided into DEX treatment group, SAP model group, SO group and 32 rats in each group; Eight rats was sacrificed at 2,6,12,24 each four time points. DEX group and SAP group using retrograde cholangiopancreatography injection of 5% sodium taurocholate(sodium taurocholate, STC)(0.1ml/100g) to build SAP model group, SO group using NS instead. Lower limb intramuscular in DEX group 0.5mg/100 g dexamethasone treatment, the remaining two groups non-intervention. Heart blood way after rats were sacrificed.(2) pancreas, liver tissue by HE staining, pancreas, liver injury pathological changes.(3) Biochemical kit used to detect serum amylase in serum.2.ELISA was used to detect the expression of NF-κB signaling pathway factor, immunohistochemistry and RT-PCR was used to detect the EPRS.(1) ELISA method to measure pancreatic tissues IκB, TLR4, INF-γ, NF-κB protein.(2) Immunohistochemistry was used to detect liver tissue EPRS protein expression.(3) Quantitative reverse transcription polymerase reaction(RT-PCR) assay the expression of the pancreas, the liver tissue EPRS mRNA.3.ELISA was used to detect the expression of NF-κB signaling pathway factor, immunohistochemistry and RT-PCR was used to detect the TNFAIP3/A20.(1) ELISA method to measure liver tissues IκB, TLR4, INF-γ, NF-κB protein.(2) Immunohistochemistry was used to detect liver tissue TNFAIP3/A20 protein expression.(3) Quantitative reverse transcription polymerase reaction(RT-PCR) assay the expression of the pancreas, the liver tissue TNFAIP3/A20 mRNA. Result1.The comparation the serum amylase enzyme and the pathological results between SO group, SAP group and DEX group(1) serum amylase were increased in group DEX and group SAP compared with group SO, group DEX serum AMS was lower than SAP group.(2) HE pancreatic pathology score shows DEX group at each time point inflammatory injury SAP group were significantly reduced(P <0.01), 24-hour time point is particularly evident. And SAP group, DEX group pancreatitis disease damage over time damage increased(P<0.05), and each time SO pancreas edema, injury no significant change. DEX group, SAP group liver pathological inflammation, damage increases with time and aggravation, peaked at 24 h time point, DEX group compared with SAP group each time to reduce the inflammatory injury(P<0.05), and each time the SO group had no significant hepatic inflammatory injury variety.2.Expression of NF-κB, IκB, TLR4, IFN-γ and the EPRS(1) DEX group was measured by ELISA test method, SAP group rat liver NF-κB, IκB, TLR4, INF-γ content at 6 hours and reached the peak point, DEX group at all time points in liver NF-κB, IκB, TLR4, INF- γ content than the SAP group decreased, while the SO group indexes do not change with time.(2) RT-PCR results are shown DEX pancreas, liver EPRS mRNA expression compared with SAP group increased, 6h DEX pancreas EPRS mRNA compared with SAP, SO were significantly higher.(3) 6h time point SO group most weak liver EPRS protein expression, and SAP group compared with weakly expressed strongly expressed mixed, DEX group the vast majority of strong expression. 6h time point SO group, SAP group and DEX group, three groups of EPRS protein expression significantly different(P<0.01); 6h time point SO group, SAP group and DEX group into three groups EPRS protein pairwise comparison, SO group and SAP There was no significant difference, but SO group and DEX group, SAP group and DEX group difference was significant, 6h DEX group EPRS protein expression compared with 6h SAP group increased expression, EPRS protein expression in three groups of rats pancreatic obvious.3.Expression of NF-κB, IκB, TLR4, IFN-γ and the TNFAIP3/A20(1) DEX pancreas NF-κB, IκB content than the SAP group lowered, SO group had no significant change. TLR4, INF-γ had no significant expression in the pancreatic tissue of each group.(2) RT-PCR results are shown DEX pancreas TNFAIP3/A20 representing SAP, SO were significantly higher.(3) TNFAIP3/A20 immunohistochemical results are shown DEX group compared with SO group, SAP group increased, and the change over time showed an increasing trend. Conclusion1. Expression of early liver EPRS occur peaked in the SAP system to activate reverse, by inhibiting the inflammatory genes involved in collaborative translation trickle thus DEX for SAP early inflammation inhibition.2. TNFAIP3 / A20 may be synergistic IkB thereby blocking NF-κB / IκB signal transduction pathway,... |
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