The Effect Of Sangpi Zhike Recipe On The TLR4/TRIF/NF-?B Signaling Pathway Of Cough In RSV-infected Mice

Purpose: In this experiment,by observing the efficacy of Sangpi Zhike Recipe in the treatment of cough BALB / c mice after RSV infection and its effect on the TLR4 / TRIF /NF-?B pathway,we explored the mechanism of Sangpi Zhike Recipe in the treatment of cough after infection.A postcough provides a strong experimental basis.Materials and Methods: A mouse model of cough after infection was established by RSV nasal infection.96 BALB / c mice were randomly divided into: normal group,model group,Sangpi Zhike Fang low dose group,mulberry The Pizhikefang middle-dose group,Sangpi Zhikefang high-dose group,and montelukast sodium group were divided into 6groups of 16 mice each,and the general state of the mice and the pathological changes of lung tissue were observed.Capsaicin stimulation was counted.After coughing,the inflammatory cells in bronchoalveolar lavage fluid were classified and counted,and TLR4 and TRIF were detected in mouse lung tissue by immunohistochemistry,Western blot,and RT-PCR.Changes in the level of NF-?B.To investigate the efficacy and mechanism of Sangpi Zhike Recipe in treating cough after infection.Results:1.Identification of the model: The number of coughs and the BALF white blood cell count of the model group were higher than that of the normal group,which was statistically significant(P <0.05);the pathology of the lungs was significantly narrower than the normal group,and the inflammatory infiltration around the airway was obvious In the normal group,no obvious pathological changes were seen.Prompt modeling success.2.General state of mice: The mice in the model group showed debilitating energy,reduced activity,dull fur,secretions in the nasal cavity and oral cavity,after the capsaicin stimulation,accelerated breathing,neck extension,abdominal muscle contraction,and scratching mouth Nasal manifestations;mice in the normal group breathed peacefully and their fur was shiny;mice in the Sangpi Zhike Recipe group and montelukast sodium group exhibited reduced activity,dull fur,secretions from the mouth and nose,and nebulization Breathing has accelerated.3.Count of cough: After the capsaicin challenge test,a small amount of cough appeared in the normal group of mice,and the number of coughs in the model mice increased significantly,which was statistically significant(P <0.05);Compared with the model group,the number of coughs in the Rustum sodium group was significantly reduced,which was statistically significant(P <0.05).3.Lung HE staining: The airway lumen of the PIC model group was narrowed,the alveolar wall was significantly thickened,and the inflammatory cell infiltration was obvious,followed by each treatment group.4.Pulmonary histopathological examination: the bronchial lumen of normal mice was smooth,the mucosal epithelium was intact,the alveolar structure was generally normal,and there was no inflammatory cell infiltration around the airway.The airway lumen of the model group was narrowed,and the inflammatory infiltration around the bronchiolar airway was obvious.No obvious pulmonary interstitial inflammation was seen.The lung pathological changes in the other drug intervention groups were improved to varying degrees.5.Count of inflammatory cells in bronchoalveolar lavage fluid: neutrophils: There was no statistical difference in bronchoalveolar lavage fluid of each group(P> 0.05).Lymphocytes,macrophages,and eosinophils: the model group was significantly higher than the normal group(P <0.05);compared with the model group,the Sangpi Zhike Fang was high,medium dose group,montelukast sodium group Decreased to varying degrees(P <0.05);the low-dose group of Sangpi Zhike Fang only reduced the white blood cell count(P <0.05).6.Immunohistochemical detection of TLR4,TRIF,NF-?B protein expression in lung tissue:Compared with the normal group,the expression of TLR4,TRIF,NF-?B protein in the model group was significantly increased,which has statistical significance(P <0.01).)Compared with the model group,the TLR4,TRIF,NF-?B protein expressions in the high-dose Sangpizhikefang group and the montelukast sodium group were significantly reduced,and the differences were statistically significant(P <0.01);compared with the model group,Compared with that,the expression levels of TRIF and NF-?B protein in the middle dose group of Sangpi Zhike Recipe decreased,and the difference was statistically significant(P<0.05).Compared with the model group,the expressions of TRIF and NF-?B protein in the low dose group of Sangpi Zhike Recipe were lower.No statistical significance(P> 0.05).7.Western blot detection of TLR4,TRIF,and NF-?B protein expression in lung tissue: Compared with the normal group,the expression of TLR4,TRIF,and NF-?B protein in the model group was significantly increased,with significant statistical significance(P <0.01);Compared with the model group,the TLR4,TRIF,and NF-?B protein expressions in the high-dose Sangpizhikefang group and the montelukast sodium group were significantly reduced,and the differences were statistically significant(P <0.01);In comparison,the expression of TLR4 and NF-?B protein in the middle dose group of Sangpi Zhike Recipe was significantly reduced,which was statistically significant(P <0.01).Compared with the model group,the expression of TLR4 and TRIF protein in the low dose group of Sang Pi Zhike Recipe was reduced,with Statistical significance(P <0.05),NF-?B expression in the low-dose group of Sangpi Zhike Fang was not statistically significant(P> 0.05).8.RT-PCR detection of TLR4,TRIF,NF-?B m RNA expression: Compared with the normal group,the expression of TLR4,TRIF,NF-m RNAB m RNA in the model group increased,the difference was statistically significant(P <0.05),compared with the model group In comparison,the expressions of TLR4,TRIF,NF-?B m RNA in Sangpi Zhike Recipe in the medium and high dose groups and montelukast sodium group were reduced,and the differences were statistically significant(P <0.05).Compared with the model group,Sangpi Zhike Recipe Low-dose group TLR4,TRIF,NF-?B m RNA expression was not statistically significant(P> 0.05).Conclusion:1.Sangpi Zhike Recipe can effectively improve the general state of mice and reduce cough symptoms.2.Sangpi Zhike Recipe can improve the pathological changes of lung tissue,significantly reduce the content of inflammatory cells in bronchoalveolar lavage fluid of PIC mice,and relieve airway inflammation.3.Sangpi Zhike Recipe can reduce the expression of TLR4,TRIF and NF-?B in the lung.It is speculated that the compound preparation may reduce the airway inflammation of cough after infection by regulating the TLR4 / TRIF / NF?B signaling pathway.