The Preventive Effect Of ChuanKeZhi On Asthmatic Mice And Its Effect On Related Abnormal Methylation Genes

Objective: To explore the preventive effect of Chuankezhi on asthmatic mice and the effect on related abnormal methylation genes.Method: Eighteen male C57BL/6J mice,6-8 weeks old,were randomly divided into 3 groups: control group,asthma group and asthmatic prevention group.The model of asthma was established by sensitization and stimulation of house dust mites.The asthmatic intervention was given before the challenge.The total number and classification of cells in the alveolar lavage fluid(BALF)of mice were observed.The IL-4 and IL-5 in BALF were detected by ELISA.TNF-? level,conventional pathological section to observe the lung inflammation,Western blot,immunofluorescence analysis of TGF-?,?-SMA,collagen I,collagen III and FGF-? and other airway remodeling related protein expression levels.T Real time RT-PCR and bisulfite genomic sequencing were used to detect the expression of related aberrant methylation genes and methylation status of promoter region.Results: Compared with the control group,there were obvious eosinophil-induced inflammatory cell infiltration and goblet cell and smooth muscle cell hyperplasia in the lung tissue and bronchi of the asthma group,while the asthmatic prevention group had obvious airway allergic inflammation reduced.The number of eosinophils,IL-5,IL-4,TNF-? in the BALF of asthma group and TGF-?,?-SMA,type I collagen,type III collagen and FGF-? in lung tissue were significantly higher than those in normal controls group(P <0.05).Real time RT-PCR analysis showed that compared with the control group,the RNA levels of pde4 d,akt1s1,pom121l2,and tm6 sf in asthma group were increased,and the RNA levels of slc8a3 and acsl3 were decreased,the difference was statistically significant(P <0.05).After asthmatic intervention,the RNA levels of pde4 d,akt1s1,pom121l2,and tm6 sf were decreased,and the RNA levels of slc8a3 and acsl3 were increased,and the difference was statistically significant(P <0.05).Promoter methylation level analysis,compared with the control group,the methylation level of pde4 d,akt1s1,pom121l2,tm6 sf promoter was decreased in the asthma group,and the promoter methylation level of slc8a3 and acsl3 was increased,of which the difference of pde4 d,pom121l2,tm6 sf,slc8a3 and acsl3 was statistically significant(P <0.05).After asthmatic intervention,the methylation levels of pde4 d,akt1s1,pom121l2,and tm6 sf promoters were increased,and the promoter methylation levels of slc8a3 and acsl3 were decreased,and the differences of pde4 d,pom121l2,slc8a3 h,and acsl3 were observed statistically significant(P < 0.05).Conclusion: Chuankezhi can prevent airway allergic inflammation and airway remodeling in asthmatic mice,which may be related to the regulation of related abnormal DNA methylation genes.