The Study Of Molecular Mechanism On The Combine Use Of Radix Et Rhizoma Rhei And Scutellariae To Treat Endotoxemia

Object:Building model by injecting LPS through tail,research based on TLR4,MAPK,and NF-κb inflammation signal pathway。Inquiry molecular mechanism on the combine use of rhubarb and scutellariae to treat endotoxemiato,and compared laboratory early research on single drug to find share advantage,provides a reliable theory on clinical using rhubarb and scutellaria.Methods:1.Animals divided based on basal body temperature and set 4h and 6h two time points for sampling observations.Rhubarb and scutellariae high doses(equivalent to raw 4.5g·Kg^-1)and low-dose(equivalent to raw 2.25g·Kg^-1),preventive drug delivery through gastric lavage,after preventive drug delivery,intravenous LPS（5mg/kg）through model,and observe the changes of organ coefficient and body temperature in rats;2.Using ELISA method and Griess assay determination rhubarb and scutellariae medicines on the rat model of sepsis 4h and 6h two time points liver and lung tissues and serum cytokine IL-1β,TNF-α,IL-6 and effects of NO content,and use qRT-PCR method to determination the corresponding m RNA expression;3.Use Western blotting detection the influence of rhubarb and scutellariae medicines on the rat model of sepsis 4h and 6h two points liver and lung TLR4-MAPKs-NF-κb signaling pathway of TLR4,p38,JNK,ERK,Iκbαand NF-κb expression of p65-associated protein and its phosphorylation.Results:1.After LPS injection molding,the body temperature of rats showed a significant two-peak phenomenon。The reason was because after rats injection of LPS,the body has a serious inflammatory response,which led to a rise in temperature.Comparing the model group,the medicine to high-low dose group can decrease the body temperature of rats induced by inflammatory reaction after LPs enters.2.Experiments found that in rats after injection of LPS model liver and lung tissues and serum cytokines TNF-and IL-1 beta,IL-6,NO significant increase in lung and liver tissue TNF-and IL-1 beta,IL-6,iNOs mRNA expression in line.Comparison group,Radix et rhizoma Rhei radix scutellariae drugs can significantly reduce the rat model of liver and lung tissue and serum TNF-and IL-1 beta,IL-6,NO content and reduced liver and lung tissues of TNF-and IL-1 beta and iNOs and IL-6 mRNA expression and we found that higher doses of inhibitory effect is obvious.3.Modeling in the liver and lung tissues in rats after TLR4 protein and gene expr ession of ERK,JNK,p38,Iκbα,NF-κb p65 protein phosphorylation levels are significantly higher,indicate that LPS in rats TLR4,MAPK,NF-Kappa b pat hways are activated.Radix et rhizoma Rhei radix scutellariae low doses can be reduc ed in animal liver and lung tissues in the rat model of liver and lung ERK,p38,Iκbα,the level of phosphorylation of NF-Kappa b p65（P<0.05）,but to model r eceptor TLR4 expression in rats did not show good effects.Conclusions:1.Radix et rhizoma Rhei radix scutellariae medicine to reduce the fever induced by LPS injection temperature,and gene and protein level reduced endotoxemia rats hepatic expression of inflammatory factors in lung tissue,and therapy of organ damage caused by LPS in rats,play a role in protection of rats.2.medicine rhubarb scutellaria drug on protection within toxins blood syndrome big rat of mechanism may is:blocking big rat body of MAPK,and NF-κb inflammation signal pathway,rhubarb scutellaria drug on can reduced big rat body ERK,and p38,and Iκbα,and NF-κb p65 this several key protein of phosphate of level to blocking whole inflammation signal of turned Guide,to reduced signal pathway downstream inflammation cell factor IL-1 beta,and TNF-α,and NO of excess expression,Our discovery of radix et rhizoma Rhei radix scutellariae on the TLR4 receptor protein inhibition is weak,with our single drug study results are very different,we need to conduct further research.