Study On The Mechanism And Function Difference Of Toll Like Receptor 2/4 Mediated By Differences In Rhabdomyolysis Acute Renal Injury

1.background and purpose:Rhabdomyolysis(RM)usually refers to the changes in the integrity of the membrane of the rhabdomyosus cells when the muscle is damaged by factors such as ischemia,inflammation,metabolism,or poisoning,and the release of the cell contents,such as myoglobin,creatine phosphokinase,and ions and small molecule toxic substances,is released into the blood.Complications such as effective circulating blood volume reduction,electrolyte imbalance,acute kidney injury(AKI)and multiple organ dysfunction syndrome.RM causes the highest incidence of AKI(RM-AKI)up to 50%.Severe cases can lead to persistent multiple organ damage and poor prognosis.The physiological and pathological mechanism of AKI has been studied clearly.Inflammation plays a key role in the process of AKI.The TOLL like receptor(TLR)is considered as the most important pattern recognition receptor,using the pattern recognition receptor(PRR)to identify the pathogen associated molecules(PAMP)of different pathogens(PAMP)and to initiate the mechanism of natural immunity.In addition to identifying the pathogen in vitro,it can also be identified by the damage related molecular model(DAMPs)to identify the injury and injury after tissue injury.Endogenous ligands produced and secreted by necrotic cells,stress cells and extracellular matrix degradation.TLR2 and TLR4 were expressed in renal tubular epithelial cells,vascular endothelial cells and mesangial cells.The purpose of this study is to observe the role of Toll like receptor 2/4 after rhabdomyolysis of acute renal injury(AKI),to analyze the difference between the two effects,and to try to explore the possible related formation mechanism through their different signal pathways,and to provide basic theoretical support for the study of acute renal injury related to inflammation and to acute renal injury.Clinical treatment provides a way of thinking.The 2.method:using the TLR2 gene knockout rat,TLR4 gene knockout rat and wild rat to establish the rhabdomyolysis AKI model,the AKI model of TLR2 and TLR4 gene knockout mice as the experimental group,the wild rat AKI model as the control group,and the serum creatinine,urea nitrogen and creatine kinase for the serum of Oh,24h and 48h.24h renal tissue,pathological PAS staining to evaluate the degree of renal injury,immunohistochemical observation of the receptor expression position,Western blot evaluation of signal pathway protein level,RT-PCR detection of inflammatory factors.SPSS 16.0 software is used for statistical analysis.3.results:(1)after the model was established,24h,wild rat(WT),TLR2 and TLR4 gene knockout mice AKI group compared with the Sham group,the serum creatinine(Cr),blood urea nitrogen(BUN)creatine kinase(CK)all increased,the model was constructed successfully.(2)compared with group WT AKI,the levels of serum creatinine(Cr)and blood urea nitrogen(BUN)in TLR2-/-AKI group and TLR4-/-AKI group decreased,while those in TLR4-/-AKI group were lowest.(3)compared with the sham group,the mice of WT,TLR2-/-and TLR4-/-AKI all had pathological injury of renal tubule,and the score of acute renal tubule injury(ATN)increased.Compared with the WT AKI group,the renal tubule injury and the ATN score decreased in TLR2-/-and TLR4-/-AKI group.(4)TLR2 and TLR4 increased in renal tubular epithelial cells and vascular endothelial cells and decreased infiltration of interstitial cells.(5)in the MYD88 signal pathway,the expression level of signal pathway protein in the TLR2-/-mice and TLR4-/-mice was lower than that in the control group,and the level of the signal pathway protein in the TLR4-/-AKI group was the lowest;in the non MYD88 signaling pathway,the expression level of the signaling pathway protein in the TLR4-/-AKI group decreased.(6)compared with WT AKI mice,the expression levels of inflammatory factors(IL-6,MIP-1 alpha,TNF-alpha,IFN-beta)in TLR2-/-and TLR4-/-mice decreased.4.conclusion:both TLR2 and TLR4 mediate RM-AKI inflammatory reaction in rhabdomyolysis acute renal injury model.Blocking TLR2 or TLR4 can effectively reduce the acute renal injury,and the effect of blocking TLR4 is more significant.TLR4 shows more inflammatory than TLR2.