Clinical Observation And Regulation Of HMGB1/TLR4/NF-?B Signal Pathway In The Treatment Of Sepsis With Anti-inflammatory Mixture

Objective To study the mechanism and futher efficacy of Anti-inflammatory decoction in the treatment of sepsis patients(heat toxin syndrome),and through cell experiments to explore the effect of Anti-inflammatory decoction in HMGB1/TLR4/My D88 signaling pathway of LPS-induced RAW264.7 inflammation model;Through the randomized controlled clinical study,to explore Anti-inflammatory decoction in sepsis patients(heat toxin syndrome)with clinical efficacy and effect on HMGB1/TLR4/My D88 signaling pathway.MethodClinical study: sepsis patients(heat toxin syndrome)were randomly divided into two groups,treatment group and control group,the treatment group was treated with integrated traditional Chinese and Western medicine: Anti-inflammatory decoction with western medicine treatment,the control group was treated with conventional western medicine treatment.To observe the before treatment and treatment after 3days and 7days changes of TCM syndrome score,blood cell analysis,liver and kidney function,IL-1,IL-6,IL-4,IL-10,TNF-?,the score of APACHE II,and ELISA has been used to determine the serum HMGB1,TLR4,My D88 levels three time points in all groups of patients.Experimental study: SD rats were administered with anti-inflammatory decoction for 7 days,to collect blood samples from the abdominal aorta and abstract drug-containing serum.Lentiviral vector carrying HMGB1 gene has been successfully constructed and maintains to silence HMGB1 RNA interference gene.RAW264.7 were divided into 7groups,blank group,LPS group,LPS+ drug-containing serum group,HMGB1+LPS group,HMGB1+ drug-containing serum+LPS group,HMGB1 RNAi+LPS group,HMGB1 RNAi+ drug-containing serum+LPS group.The expression of TLR4 m RNA,My D88 m RNA,NF-?B m RNA in each group was determine by RT-PCR and the level of IL-6,IL-10 and TNF-? in supernatants of cell were determine by ELISA.Result Clinical study: 7 days after treatment,the clinical efficacy of the treatment group was significantly better than that of the control group.(P<0.05).The total effective rate of the treatment group was 93.103% and that of the control group was 78.571%.The total scores of TCM syndromes,constipation,fear heat and abdominal distention in the treatment group were significantly lower than those in the control group(P < 0.05),and The proinflammatory factors IL-1,IL-6 and TNF-? in the treatment group were significantly lower than those in the control group 3 and 7 days after treatment.(P < 0.05).anti-inflammatory factors IL-4 and IL-10 were significantly higher than those in the control group(P < 0.05);HMGB1,TLR4 and My D88 levels in the treatment group were significantly lower than those in the control group(P < 0.05);APACHE II scores in the treatment group were significantly lower than those in the control group(P < 0.05).Experimental study: The levels of IL-6 and TNF-? in LPS group were higher than those in blank group(p<0.05).The levels of IL-6 and TNF-? in LPS+ drug-containing serum group,HMGB1+ drug-containing serum+LPS group and HMGB1 RNAi+ drug-containing serum+LPS group were lower than those in LPS group,HMGB1+LPS group and HMGB1 RNAi+LPS group(p<0.05).The levels of IL-6 and TNF-? in HMGB1+LPS group were higher than those in LPS group,The levels of IL-6 and TNF-? in HMGB1 RNAi+LPS group were lower than those in LPS group(p<0.05);The levels of IL-10 in LPS group were higher than those in blank group(p<0.05);The levels of IL-10 in LPS+ drug-containing serum group,HMGB1+ drug-containing serum+LPS group and HMGB1 RNAi+ drug-containing serum+LPS group were higher than those in LPS group,HMGB1+LPS group and HMGB1 RNAi+LPS group(p<0.05);The levels of IL-10 in HMGB1+LPS group were higher than those in LPS group,The levels of IL-10 in HMGB1 RNAi+LPS group were lower than those in LPS group(p<0.05);the expression of TLR4 m RNA,My D88 m RNA and NF-?B m RNA of LPS group were higher than those in blank group(p<0.05);the expression of TLR4 m RNA,My D88 m RNA and NF-?B m RNA of LPS+ drug-containing serum group,HMGB1+ drug-containing serum+LPS group and HMGB1 RNAi+ drug-containing serum+LPS group were lower than those in LPS group,HMGB1+LPS group and HMGB1 RNAi+LPS group(p<0.05).the expression of TLR4 m RNA,My D88 m RNA and NF-?B m RNA of HMGB1+LPS group were higher than those in LPS group,the expression of TLR4 m RNA,My D88 m RNA and NF-?B m RNA of HMGB1 RNAi+LPS group were lower than those in LPS group(p<0.05).Conclusion Clinical study: the curative effect of TCM syndrome of treatment group were better than the control group;the TCM syndrome of treatment group were significantly better than control group(fever,constipation,fear heat and abdominal distention);Therefore,Anti-inflammatory decoction in the treatment of sepsis patients(heat toxin syndrome)has good clinical curative effect;It can down-regulate the levels of HMGB1,TLR4 and My D88,down-regulate pro-inflammatory factors and up-regulate anti-inflammatory factors.It plays a protective role on the body.Experimental study: Anti-inflammatory decoction has protective effects on LPS-induced RAW264.7 inflammation model,and its mechanism may be related to its down-regulation of HMGB1,TLR4,My D88 and NF-kappa B,up-regulation of anti-inflammatory factors,down-regulation of pro-inflammatory factors,regulation of immune response,and alleviation of cell damage caused by inflammatory reaction.