| objective:Bronchial asthma,which is the most common and multiple respiratory diseases,seriously impacts on human health and quality of life.Foreign-related studies have shown that the silent information regulator protein 1(SIRT1)was closely related with asthma,significantly higher in the asthma model mice than control group.While sirtinol able to reduce the SIRTI level,but the exact strength of inhibition is not clear.This experiment investigated the effect of different concentrations sirtinol on the 1 SIRT1 gene expression and interleukin 5(IL-5)levels of asthma,to find the optimal concentration of its role,and compared the various concentrations of sirtinol to dexamethasone on asthma therapeutic effect,to explore new ways to provide theoretical basis for the treatment of asthma.Methods:42 Balb/c female mice were randomly divided into blank control group(Bk group)(n = 6),dimethyl sulfoxide control group(Md group)(n = 6),asthmatic model group(As group)(n = 6),the dexamethasone intervention group(Dm group)(n = 6),the sirtinol intervention groups(S1,S2,S3 group)(n =6*3).Asthma model group:each mouse was sensitized by intraperitoneal injection of 0.2ml antigen suspension(chicken ovalbumin OVA100ug + aluminum hydroxide 2mg + saline 0.2 ml)in the first and the 8th and the fifth day.To inspire by intranasally instilled the 40ulOVA solution(2gOVA +100 ml normal saline)into each mouse after 0.75%pentobarbital anesthesia on the 16th day,and repeated it for 7 days.Dexamethasone intervention group:dexamethasone solution 0.2ml(3mg/kg)had been intraperitoneally injected 30 minutes before stimulation and the other was the same as asthma group.The sirtinol intervention group(S1,S2,S3 group):sensitized as asthmatic model group,the sirtinol solution was injected intraperitoneally twice separately 1 hour before the first challenge and three hours after the last.In S1 group,the intraperitoneal injection of sirtinol concentration was 0.05mg/kg.The sirtinol concentration was 0.5mg/kg in S2 group.In the S3 group was 3.0mg/kg.Blank control group:each mouse was sensitized by intraperitoneal injection of 0.2ml a saline in the first and the 8th and the fifth day.To inspire by intranasally instilled the 40ul normal saline into each mouse after 0.75%pentobarbital anesthesia in the 16th day,and repeated it for 7 days.Dimethyl sulfoxide control group:the Sensitization and intraperitoneal injection time just as the sirtinol intervention group,the intraperitoneal injection of 0.125%dimethyl sulfoxide(DMSO)solution as the solvent control group.All mice were sacrificed 24 hours after the last challenge.Assess the general morphology and airway inflammatory cell infiltration of the lung tissue of mice by HE staining.Detect the expression of SIRT1 in lung tissue by immunohistochemistry.Line the SIRT1 and reference gene(ACTB)mRNA levels in lung tissue of mice using real-time fluorescence quantitative PCR.Detect the IL-5 levels of alveolar fluid(BALF)by sandwich enzyme-linked immunosorbent assay(ELISA).All data was expressed like x±s.SPSS 13.0 software systems for processing statistical data,p<0.05 indicates a significant difference.ANOVA analytic method was used between different groups.Using linear correlation analytic method,analysis the correlation between SIRT1 expression level in the lung tissue and the IL-5 in BLAF. Results:(1)Lung tissue histopathology HE staining:In asthma model group,a large number of inflammatory cells infiltrated around the small airways and small blood vessels.The edema of the mucosa and submucosa can be seen.There were eosinophils,monocytes,and the overflow of red blood cells visible in the alveolar space.Alveolar septum fractured.In blank group,the lumen was smooth without occlusion.And there was no tracheal mucosal edema,no infiltration of inflammatory cell around the small airways and small blood vessels.There can be seen a large number of inflammatory cell infiltration In dmethyl sulfoxide control group.In dexamethasone interfere group,there were less inflammatory cells and exudate.Inflammation significantly reduced.The alveolar structure of the S1 group was clear.In alveolar space and alveolar interstitial,inflammatory cells and exudates reduced.And inflammation was sharply absorbed.S2 group absorbed partly.And S3 was a small amount of inflammation absorption.(2)Immunohistochemical detection of lung organization SIRT1 expressions:The SIRT1 expression in Bk group lung tissue airway epithelial was negative.As group and the Md group SIRT1 expression was strongly positive.Group Dm expression was weakly positive.In S1,S2,S3 lung tissue,airway epithelial SIRT1 expression were positive,in which the S1 group was the weakest,the S3 group the strongest,S2 between the two.(3)Fluorescence quantitative PCR detection of lung tissue SIRT1mRNA expression:?the highest SIRT1 mRNA expression level was in the As group and the Md group lung tissue,the difference was not significant(P>0.05)between them.The lowest was in the Bk group,and the others were between the two levels.?Compare to As group,the Dm group SIRT1 expression levels were significantly lower(P<0.05).?Relatively As group SIRT1 level of sirtinol intervention group were decreased.With the increase of sirtinol concentration,its expression levels increased.The S1 group was the lowest,S3 the highest,and S2 living between the two.Compared with each group,there were significant differences(P<0.05).?SIRT1 expression levels of the S1 group,and the Dm group,the difference was not statistically significant(P>0.05).While Dm group differed significly from S2 and S3(P<0.05).(4)IL-5 in Bk group BALF was the lowest.The As group and Md IL-5 content were the highest,and the difference was not significant(P>0.05).Other groups were between them.Relative As group,IL-5 levels of Dm group was significantly reduced,and the difference was statistically significant(P<0.05).Otherwise in the sirtinol intervention group,the levels were lower to As group.But with sirtinol concentration increased,IL-5 levels showed an increasing trend,the S1 group had the lowest,S3 the highest,and S2 between them,two pairwise comparisons were significantly different(P<0.05).Dm group and the S1 group of IL-5 levels were no significant difference(P>0.05).The Dm group differed sharply from S2 and S3(P<0.05).Conclusions:(1)Compared with blank group,the SIRT1 expression was significantly increased in asthma model mice.Otherwise dexamethasone can significantly reduced it in asthma mice.(2)The inhibition of SIRT1,sirtinol,inhibited SIRTI's expression levels of lung tissue and IL-5 level in BALF effectively in the low concentration.With the concentration increased,the anti-inflammatory role would be weakened.(3)Of dexamethasone and sirtinol,there was no difference in the role of lower levels of asthma in mice of SIRT1.But the latter was connected with the concentration.(4)The level of SIRT1 expression in lung tissue was positively correlated with IL-5 levels in BLAF.So in allergic airway diseases,SIRT1 can suggest the degree of airway inflammation. |
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