Effects Of The Method Of Benefiting Qi To Relieve Superficies Syndrome On ?-defensin-2?Caspase1 In Lung Tissue Of "Lung Qi Deficiency With Exogenous Cause" Rats Model And Its Correlation With Pyroptosis

ObjectiveTo explore the influence of shensu Drink on ?-defensin-2?Caspase1 in lung tissue of “lung qi deficiency with exogenous cause” rat model and the relationship between lung qi deficiency with exogenous cause and Pyroptosis.Methods1.Copy the “Lung qi deficiency with exogenous cause”rats model by using the united method of “smudge + cold stimulation +LPS injection in trachea”.Monitoring the change of general condition?body weight growth of normal rats and model rats during the whole process;Take every rats' lung?spleen and thymus after executed them,weight and measure and calculate each groups' different in rats' weight,lungs' ?spleens' and thymus' visceral index;And the histomorphology differences in the lung tissue of two groups rats.In general,the differences above of the two group rats will be the criteria of successfully copied the “Lung qi deficiency with exogenous cause”rats model in this project.2.Set Shensu Drink as the carrier of the method “benefiting qi to relieve superficies syndrome”.Intragastric administrate “Lung qi deficiency with exogenous cause” rats model with different dose of Shensu Drink and positive control drug,take away and homogenate every rats' lung tissue after execute them,detect and analyze the influence of Shensu Drink in the content of IL-1? ? IL-18 and BD-2 in lung homogenate of each group via the method of ELISA.3.Measure and calculate the influence of Shensu Drink in the expression of Caspase1 mRNA?NLRP3 mRNA in lung tissue of each group via the method of Real-time fluorescent quantitative PCR(RT-PCR).4.Measure and calculate the influence of Shensu Drink in the expression of protein of rBD-2 ? NF-?B p65 in lung tissue of each group via the method of immunohistochemical technology.Results1.The united method of “smudge + cold stimulation + LPS injection in trachea” successfully copied the “Lung qi deficiency with exogenous cause”rats modelModel rats successively occurred less activity ?hairs become yellow and drop?the body weight growth slowed(P<0.05)?defecate comes to be rare and some of them appeared wheezy phlegm(moist rales);T he visceral index of lung?spleens and thymus of model rats lower than normal rats' obviously(P<0.05);Pathological section of model rats appeared : Pulmonary bronchiallumen expansion at all levels,Mucosal epithelial necrosis fall off,pseudostratified ciliated columnar epithelium large depigmentation,vessels expand andcongest,a large number of inflammatory cells infiltration,pulmonary interstitial inflammatory cell infiltration,Submucosal wall looseorganization and edema,Alveolar walls obviously thickened,the alveolar walls of part of the animal fractured,fused,formed mild emphysema.2.The influence of Shensu Drink on the content of IL-1??IL-18 and?-defensin-2 in lung homogenate of rats in each groupCompared with the blank group,the content of IL-1??IL-18 and ?-defensin-2 in lung homogenate of the model group were significantly increased(p<0.05);Compared with the model group,the content of IL-1? in lung homogenate of the high-dose group /middle-dose group /low-dose group of Shensu Drink and the positive control group were significantly decreased(p<0.05);The contentof IL-18 in lung homogenate of the middle-dose group and low-dose group of Shensu Drink has significantly decreased(p<0.05)when compared with the model rats;The contentof?-defensin-2 in lung homogenate of the high-dose group /middle-dose group/low-dose group of Shensu Drink has significantly increased(p<0.05)when compared with the model rats;The content of IL-1? in lung homogenate of the high-dose group/middle-dosegroup/low-dose group of Shensu Drink and the positive control group had no significant difference(p>0.05);The content of IL-18 in lung homogenate of the high-dose group /middle-dosegroup/low-dose group of Shensu Drink significantlydifferent from each other(p<0.05);The content of ?-defensin-2 in lung homogenate of the high-dose group /middle-dosegroup/low-dose group of Shensu Drink had no significant difference(p>0.05).3.The influence of Shensu Drink on the expression of Caspase1 m RNA?NLRP3m RNA in lung tissue of “lung qi deficiency with exogenous cause” rats.Compared with the blank group,the content of Caspase1 mRNA?NLRP3 mRNA that expressed in lung homogenate of the model group rats were significantly increased(p<0.05);Compared with the model group,the content of Caspase1mRNA?NLRP3 mRNA that expressed in lung homogenate of the high-dose group/middle-dose group /low-dose group of Shensu Drink and the positive control group were significantly decreased(p<0.05);The content of Caspase1 mRNA?NLRP3mRNA that expressed in lung homogenate of the high-dose group/middle-dosegroup/low-dose group of Shensu Drink and the positive control group had no significant difference(p>0.05);4.The effect of Shensu Drink on the content of protein rBD-2?NF-?B p65 that expressed in lung tissue of “lung qi deficiency with exogenous cause” rats.Compared with the blank group,the content of protein BD-2,NF-?B p65 that expressed in lung tissue of the model group rats were significantly increased(p<0.01);Compared with the model group,the content of protein BD-2 that expressed in lung tissue of the middle-dose group of Shensu Drink and the positive control group were significantly decreased(p<0.05);The content of protein NF-?B p65 that expressed in lung tissue of the high-dose group of Shensu Drink and the positive control group were significantly decreased(p<0.01).Conclusions1.Shensu Drink could inhibit the secretion of the NLRP3 inflammasomes' downstream inflammatory factors IL-1? and IL-18 in the inflammatory pathological process of “lung qi deficiency with exogenous cause” rats.Shensu Drink may has a bidirectional regulation effect on the expression of BD-2 in the lung tissue of“Lung qi deficiency with exogenous cause” rats,which is likely to be related to the signal pathway of NF?B.2.Pyroptosis mediated by Caspase-1 participated in the inflammatory pathological process of “lung qi deficiency with exogenous cause” rats.Shensu Drink could inhibit the occurrence of Pyroptosis that mediated by Caspase-1,and its may be one of the mechanisms of Shensu Drink exerting the “benefiting qi to relieve superficies syndrome” efficacy.