The Intervention Effect Of Angong Niu Huang Wan On The JAK/STAT Pathway Of Sepsis Rats

Objective:based on successful replication of sepsis animals, the paper studies on the pathogenesis of sepsis from the level of molecular cell. Using the intervene treat of Angong Niu Huang Wan, the paper will observes how the pill protects the septic rats. On the basis of that, it also will check the pills'mRNA expression of early sepsis inflammatory media TNF-aand later media HMGB-1, also its effect of Janus kinase/signal transducer and activators of transcription (JAK/STAT). The pape r will offer the theoretical foundation for the application of Angong Niu Huang Wa n in injury severity from exploring the mechanism of how Angong Niu Huang Wan treating the sepsis on the basis of molecular mechanism, and deep level target.Methods:adopting CLP to replicate the sepsis animal model; 90 SPF rats, hal f male and half female, divided into five groups, (1) the control (n=10), get the blo od after anesthesia then euthanize them for the test; (2) the model (n=20), get the blood 24h later after treating the rats with distilled water (10ml/kg) 0.5h before CL P and 6h after CLP, then euthanize them for the test; (3) Angong Niu Huang Wan low, middle and high, each 20 rats, get the blood 24 hours after gastric perfusion (1.0g/kg,2.0g/kg,3.0g/kg),0.5h before CLP and 6h after CLP, then euthanize them for the test; observe each groups and check the death rate and normal behaviors d uring the 24h; the degree of animal autopsy intraperitoneal infection; the rats rectal temperatures 6h,12h and 24h after operation; the WBC of the rats 24h after operati on measured by blood analyzer; the ALT, AST, BUN, Crea, TB levels of the Seru m measured by automatic biochemistry analyzer; the MPO content in the lung meas ured by enzymatic determination; the toxin content in the plasma by limulus polyph emus; the expression of STAT1, STAT3, TNF-αand HMGB-1 of liver, lung and kidney measured by semi-quantitative RT-PCR; the DNA binding activity of STAT1 and STAT2 measured by EMSA.Results:based on replication of sepsis animal models by CLP, the ill conditio ns of the rats gradually increased and some rats would even die; the activities of ra ts were induced and they no longer gather together to keep warm; they has the feat ures of sleepiness with pio-erection and do not drink water; some rats even suffer t he shortness of breath, pyuria, diarrhea and eye exudate. The cumulative rate of dea th reaches 55% in 24h. Post mortem examination reveals the serious degree of intra peritoneal infection; the models'body temperatures keep in a low condition (core te mperature less then 36℃), get leukopenia (WBC maybe less than 4×109/L) and the ir main organs are seriously damaged (ALT, AST and TB levels of serum and lung MPO activity are obviously higher than the control group); the toxin level in the s epsis rats'serum is very higher than the control group; the DNA activity of STAT1 and STAT 3 of the sepsis rats'main organs also rises obviously, JAK/STAT is al so highly activated in the pathway and meanwhile, the expression of early inflamma tory media TNF-αand later media HMGB-1 is also highly raised.Angong Niu Huang Wan can improve the normal behaviros of the sepsis rat s, obviouly reduce their death rate, raise their tempeatures during their shock and in crease their WBC count. The pills can lower the ALT and AST levels in the sepsis rats'serum, higly decreased their TB level in the serum, their lungs'MPO activity and the toxin content in their plasma. The pills can reduce JAK obviously. STAT1 and STAT3's mRNA expression in the STAT activation pathway can reduce STAT s'RNA bending activity, the pathway's activation degree of JAK/STAT and meanw hile the Mrna expression of TNF-αand HMGB-1.Conclusion:the CLP method can successfully replicate the sepsis rate models; the pathogenesis of the septic rats might be related to the higher pathway activatio n of JAK/STAT and the release of early inflammatory media TNF-αand later med ia HMGB-1 caused by the activatio pathway so as to cause the inflammatory effect s out of control and finally lead to the organs damage, MODS and septic shock. A ngong Niu Huang Wan can well protect the sepsis. The possible molecular mechani sms treatment of the pills for sepsis is to reduce the STAT1 and STAT3 expression in the JAK/STAT activation pathway so as to reduce the JAK and STAT's activati on pathway degree, thus reduce the express of related cytokine such as TNF-α, H MGB-1, from JAK/STAT activation pathway, regulate the inflammatory infection tha t is out of control and reduce the inflection damage to organs so as to protect orga nism.