Effect Of Liuwei Buqi Recipe On Airway Inflammation And TLR4/NF-κB Signal Pathway In COPD Model Rats

PurposeTo study the effect of Liuwei Buqi recipe on airway inflammation and TLR4/NF-κB signaling pathway in COPD model rats and its possible mechanism of action,and try to reveal the possible target,links and pathways for the treatment of COPD,which is a further clinical step.Optimize the treatment of COPD to obtain evidence.MethodsChoose 60 male SD rats,select 10 as the normal control group,The rest of the rats make a COPD model.The model group rats in the first day and 14 th day of experiments use the trachee to add fat and lipopolysaccharide(LPS)smoked double factors to establish a COPD experimental animal model,and the modeling cycle is 3 months.Rats after successful model were divided into 5 groups according to the random digital table:the model control group,Guilong Kechuanning group and Liuwei Buqi low,middle and high dose group,10 animals each group.The normal control group and the model control group gifted 10ml/kg/d physiologic saline,Guilong Kechuanning group gavaged0.82g/kg/d Guilong Kechuanning suspension,Liuwei Buqi low,middle and high dose group gavaged0.265g/kg/d,0.53g/kg/d,1.06g/kg/d Liuwei Buqi suspension.It started administration intervention at the first day on the 9th week of manufacturing,2 times a day,the course of treatment was 12 weeks.Record the general conditions of each group and respiratory symptoms;measure lung function after intervention;HE staining detected pathological changes in lung tissue;TLR4 and NF-κB expression were detected by immunohistochemistry;ELISA method was detected by TNF-α in serum and BALF IL-1β,IL-4,IFN-γlevel;Western blot measuring TLR4,NF-κB,IκBα protein expression;RT-q PCR method for detection of TLR4,NF-κB,My D88 m RNA level.Results1.The general conditions and respiratory symptoms:The normal control group of rats breathing,spirit,fur,reaction,diet,the amount of water,and the second sleeve are normal;compared with the normal control group,the model control group has a cough,breathing frequency is faster,obviously breathing,audible and breathing sound,frequently grasping the nose,hairless,partial hair loss,slow response,irritability,sweat,filling water supply;compared with the model control group,the general situation of Guilong Kechuanning group and Liuwei Buqi low,middle and high dose group were significantly improved,Among them,the improvement of Liuwei Buqi high dose group is most obvious.The degree of improvement in Liuwei Buqi middle dose group and Guilong Kechuanning group is quite,the improvement effect of Liuwei Buqi low dose group is lower than Guilong Kechuanning group.2.Lung function:Compared with the normal group,the model group the FVC,FEV0.3,FEV0.3/FVC(%)and PEF values of rats decreased significantly(P<0.05);compared with the model control group,Guilong Kechuanning group and Liuwei Buqi low,middle and high dose group were increased(P<0.05),among them,Liuwei Buqi high dose group is most significant;It has statistically significant significance compared to the Guilong Kechuanning group and the Liuwei Buqi low,middle dose group(P<0.05);the degree of lung function improvement in Kechuanning group is more no statistics in the Liuwei Buqi middle dose group significance(P>0.05),has statistically significant with the Liuwei Buqi low dose group(P<0.05).3.Pathological of lung tissue:The bronchial wall structure of the normal control group was complete,no obvious inflammatory exudate;compared with the normal control group,The bronchial wall of the model control group and the surrounding lung tissue have a large amount of inflammatory cells that are mainly lymphocytes,macrophages,and the alveolar cavity is irregular,and some alveolar is broken into pulmonary bulks;compared with the model control group,the pathological pathology of lung tissue in each dose group of Guilong Kechuanning group and the Liuwei Buqi is different degrees,and the inflammatory cell infiltration of airway is alleviated.Among them,Liuwei Buqi high dose group is the most obvious;The degree of improvement in Guilong Kechuanning group is quite to Liuwei Buqi middle dose group,and the degree of improvement in the two groups is better than Liuwei Buqi low dose group.4.Expression of TLR4 and NF-κB in lung tissue:The lung tissue TLR4 and NF-κB in the normal control group did not express;compared with the normal control group,the number of TLR4 and NF-κB positive cells in the model control group was more,which showed strong positive expression;compared with the model control group,The number of TLR4 and NF-κB positive cells in rats lung tissue in each dose group in Guilong Kechuanning group and the Liuwei Buqi are reduced,and the expression is weakened,among them,the highest expression of Liuwei Buqi high dose group and is the most weak;Liuwei Buqi low dose group TLR4 and NF-κB were moderately expressed,and in Guilong Kechuanning group and Liuwei Buqi middle dose group is moderately or weakly positive.5.Serum and BALF levels of TNF-α,IL-1β,IL-4,IFN-γ:Compared with the normal control group,the serum and BALF levels of TNF-α,IL-1β,and IL-4 in the model control group were increased,IFN-γlevels were decreased(P<0.05);compared with the model control group,Guilong Kechuanning group and Liuwei Buqi each dose group,serum and BALF levels of TNF-α,IL-1β and IL-4 decreased significantly,IFN-γlevels increased significantly(P<0.05),among them,the Liuwei Buqi high dose group has the most significant changes;It has statistically significant significance compared to Guilong Kechuanning group and Liuwei Buqi low,middle dose group were statistically significant(P<0.05);The serum and BALF in Guilong Kechuanning group,IFN-γ,TNF-α,IL-1β,IL-4 level were more no statistically significant in Liuwei Buqi middle dose group(P>0.05),with Liuwei Buqi low dose group was compared to statistically significant(P<0.05).6.Protein expression of TLR4,NF-κB,IκBα:Compared with the normal control group,the protein expressions of TLR4,NF-κB,and IκBα in the model control group were significantly increased(P<0.05);compared with the model control group,Guilong Kechuanning group and Liuwei Buqi low,middle and high dose group were decreased(P<0.05),among them,the decline in Liuwei Buqi high dose group is most significant,and it has statistically significant compared with Guilong Kechuanning group and Liuwei Buqi low,middle dose group(P<0.05);the expression of TLR4,NF-κB,and IκBα protein expression in Guilong Kechuanning group will have no statistical significance in Liuwei Buqi middle dose group(P>0.05),which is more statistically significant with Liuwei Buqi low dose group(P<0.05).7.Levels of TLR4,NF-κB,My D88 m RNA in lung tissues:Compared with the normal control group,the levels of TLR4,NF-κB and My D88 m RNA in the model group increased significantly(P<0.05);compared with the model control group,Guilong Kechuanning group and Liuwei Buqi low,middle and high dose group were significantly decreased(P<0.05),among them,Liuwei Buqi high dose group decreased was most significant,with Guilong Kechuanning group and Liuwei Buqi low,middle dose group was statistically significant(P<0.05);the level of TLR4,NF-κB and My D88 m RNA in Guilong Kechuanning group will have no statistical significance in Liuwei Buqi middle dose group(P>0.05),it has statistically significant comparison with Liuwei Buqi low dose group(P<0.05).Conclusion1.Liuwei Buqi recipe can reduce the expression of TLR4 and NF-κB in the lung tissueof COPD rats,reducing serum and BALF levels in inflammatory factors,improving the pathological changes of lung tissue,improving lung function.2.The intervention effect of Liuwei Buqi recipe on airway inflammation in COPD rats may be achieved by regulating the TLR4/NF-κB signaling pathway.