| ObjectiveTo investigate the effects of Yanghe pingchuan Granules on immune function and airway inflammation in asthmatic model rats.MethodsForty-eight SPF-grade SD male rats were randomly divided into six groups,namely normal control(group N),model group(group M),Yanghe pingchuan granules low-dose group(group YHL),Yanghe pingchuan granules medium dose group(group YHM),Yanghe pingchuan granules high dose group(group YHH)and dexamethasone group(group DXM),with eight rats in each group.The rat model of asthma was established by using OVA combined with aluminium hydroxide twice for sensitization and daily stimulation for 4 weeks.Intervention method:Yanghe pingchuan granules low,medium and high dose groups were gavaged with 3.87,7.74 and 15.48g/(kg-d)of Yanghe pingchuan granules suspension once a day respectively;dexamethasone group(DXM group)was gavaged with 0.125mg/(kg·d)of dexamethasone suspension once a day;The normal control group(group N)and model group(group M)were given 0.9%Na Cl injection 1ml/(100g-d)by gavage.On the day of stimulation,the corresponding concentration of drug was administered 1h before stimulation.During the whole experiment,the rats in each group were observed for their feeding and drinking patterns,body weight gain,mental status,hair and limb damage;during the sensitization and excitation period,the rats in each group were observed for any allergic reactions such as sneezing,scratching,wheezing and shortness of breath.The histomorphological changes of lung and spleen tissues of the rats were observed by HE staining.The inflammatory cells in the alveolar lavage fluid of each group of rats were classified and counted by Jimsa staining.The percentage of peripheral blood T-cell subsets(CD3~+,CD4~+,CD8~+)was measured by flow cytometry,and the CD4~+/CD8~+ratio was calculated.Serum IgE,IgG,IgM and Th1,Th2 cytokines IL4 and IFN-γwere detected by ELISA.Results1.General situation of rats:rats in group N had regular diet water intake,bright hair color and regular weight increase.Compared with group N,after sensitization,the rats in group M had reduced dietary intake,raised hair,dark yellow color,poor mental state,restlessness,slow weight gain,wheezing,shortness of breath,sneezing,nose scratching and other symptoms.After intervention,the general conditions of rats in each dose group of Yanghe pingchuan granules were improved to varying degrees,and asthma symptoms such as wheezing,shortness of breath,sneezing and nose scratching were significantly improved.2.HE staining results of lung tissues of rats in group N were structurally intact,with smooth bronchial walls and insignificant inflammatory cell infiltration.In group M,the lung tissues were severely damaged,with thickened bronchial walls,narrowed lumen,and massive inflammatory cell infiltration.In group YHL,the lung tissues were mildly damaged,with thickened bronchial walls,narrowed lumen,and significant inflammatory cell infiltration.In group YHM,the lung tissues were less structurally damaged than in group M,with mildly thickened bronchial walls,narrowed lumen,and reduced inflammatory cell infiltration.The lung structure of the rats in the YHH group was more intact,and the thickening of the bronchial walls and narrowing of the lumen were reduced significantly,with a small amount of inflammatory cell infiltration.3.HE staining results of spleen tissue showed that the red pulp and white pulp in spleen tissue of rats in group N were clearly delimited and evenly distributed.The margin areas of the red pulp and white pulp in the spleen tissue of rats in group M were widened,blurred and unevenly distributed,the white pulp hyperplasia was obvious,and the spleen nodules were seriously damaged,indicating the proliferation of lymphocytes.The red pulp and white pulp of YHL group were blurred and unevenly distributed,and the hyperplasia of white pulp lymphatic area was slightly reduced than that of M group.In YHM group,the red pulp and white pulp were blurred and unevenly distributed,the spleen nodules were relatively complete,and the hyperplasia in the lymphatic area of white pulp was significantly reduced compared with that in M group.In YHH group,the red pulp and white pulp were clearly demarcated and evenly distributed,and the lymphatic area of white pulp was slightly hyperplasia.In DXM group,the red pulp and white pulp were clearly demarcated and evenly distributed,and the lymphatic area of white pulp was slightly hyperplasia.4.Compared with N group,the count of Eos,Neu and Lym in alveolar lavage fluid of rats in M group was significantly increased(P<0.01);Compared with M group,the counts of Eos,Neu and Lym in the alveolar lavage solution of rats in Yanghe pingchuan granule groups and DXM groups were decreased(P<0.01).the YHH group had the best effect among all dose groups of Yanghe pingchuan granules(P<0.01).5.Compared with group N,peripheral blood CD3+,CD4+cell percentage and CD4+/CD8+ratio in group M increased to different degrees,and CD8+cell percentage decreased significantly(P<0.05);compared with group M,peripheral blood CD3+,CD4+cell and CD4+/CD8+percentage in rats in YHH and DXM groups decreased significantly(P<0.05 or P<0.01),and the percentages of CD8+cells were significantly higher in each dose group of Yanghe pingchuan granules and DXM group(P<0.01).6.Compared with group N,the serum IgE level of rats in group M increased significantly and IgG and IgM levels decreased significantly(P<0.01);compared with group M,the IgE levels of rats in all dose groups of Yanghe pingchuan and DXM group decreased significantly(P<0.01),and the serum IgG and IgM levels of rats in both YHH and DXM groups increased significantly(P<0.01).7.Compared with group N,serum IL4 level was significantly increased and IFN-γlevel was decreased in group M(P<0.01);compared with group M,serum IL4 level was decreased(P<0.01)and IFN-γlevel was increased in both YHH and DXM groups to different degrees(P<0.01).8.The percentage of peripheral blood CD3+,CD4+cells and CD4+/CD8+ratio were positively correlated with serum IL4 levels and IL4/IFN-γratio(P<0.05 or P<0.01),while the percentage of CD8+cells was negatively correlated with IL4 levels and IL4/IFN-γratio(P<0.01)in all groups of rats.The percentage of CD3+and CD4+cells and CD4+/CD8+ratio in peripheral blood were negatively correlated with serum IFN-γlevels(P<0.05)and the percentage of CD8+cells was positively correlated with serum IFN-γlevels(P<0.05).9.Serum IgE levels were positively correlated with IL4 levels and IL4/IFN-γratio in all groups of rats(P<0.01),IgG and IgM levels were negatively correlated with IL4levels and IL4/IFN-γratio(P<0.01).IgE levels were negatively correlated with IFN-γlevels(P<0.01),IgG and IgM levels were positively correlated with IFN-γ(P<0.01).Conclusion1.Yanghe pingchuan Granules can improve the general condition of rats and reduce the inflammatory cell infiltration of lung tissue.2.Yanghe pingchuan Granules can improve the immune function of asthmatic rats by spleen immunity,cellular immunity,humoral immunity and other aspects.3.Yanghe pingchuan granules may mediate Th1/Th2 imbalance by regulating immune function,thus reducing airway inflammation of asthma. |
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