| Objective:To explore the effect of Chaipu Decoction on expression of GR-?,GR-? and AP-1 in mouse of steroid-resistant asthma.Methods: Lipopolysaccharide(LPS)was used to induce steroidresistant asthma in a murine model sensitized and challenged with ovalbumin(QVA).30 6-week-old female BALB/c mice were randomly divided into 5 groups of normol group,SRA group(hormone resistance group),SRA+dexamethasone group(dexamethasone group),SRA+Chaipu Decoction group(Chaipu Decoction group),SRA+ dexamethasone+Chaipu Decoction group(dexamethasone +Chaipu Decoction group),(n=6 each).At Day 1 and 14,the normol group was received an intraperitoneal injection of nomal saline 0.1ml,the rest of four grous had an intraperitoneal injection of a mixture of OVA 20?g+Al(OH)3 2.25mg+normal saline to 0.1ml.At Day 27,the normal group was intranasally instilled normol saline(20ul)while the rest of four groups had an intranasal instillation of LPS(10ug).AT Day 27,nomal group,hormone resistance group and dexamethasone group were intragastric administration with normol saline while Chaipu Decoction group and dexamethasone +Chaipu Decoction group were intragastric administration with Chaipu Decoction.At Days 28,29 and 30,the normal group was challenged via airway with normal saline and the rest of four groups with 1% OAV in normal saline.The normal group,hormone resistance group and Chaipu Decoction group were intraperitoneally injeccted with normal saline 30 min pre-challenge while dexamethasone group and dexamethasone +Chaipu Decoction group with dexamethasone(3mg/kg).The normol group,hormone resistance group,dexamethasone group received intragastric administration of normal saline 1h pre-challenge;Chaipu Decoction group,dexamethasone +Chaipu Decoction group received intragastric administration of Chipu Decoction 1h pre-challenge(1.5g/kg).The rats were sacrificed by intraperitioneal anesthes 24 hours after the last nebulization and blood was exsanguinated from the abdominal aorta.The right lung tissue of rate was examined by HE stanining and immunohistochemistry to detect the protein expression of GR-?,GR-? and AP-1,the left lung GR-?,GR-?,AP-1 mRNA levels.Results: 1.The changes of histopathologicalHE staining showed that there were no airway inflammatory changes in the lung tissue of normal control gr.The lumen struture was regulation,airway wall was no thickening.The average thickness of airway in hormone resistance group was significantly increased,bronchial wall layers were observed in a large number of inflammatory cells,hyperplastic mucus gland and goblet cells leaded to excessive secretion of mucus in the lumen.Compared with hormone resistance group,the airway lesions were reduced in the both of the dexamethasone group and Chaipu Decoction group,but there was no singnificant difference between the two groups.The dexamethasone +Chaipu Decoction group had singnificantly reduced bronchial lesions compared with hormone resistance group.2.The expression of GR-?,GR-?,AP-1 protein in lung tissue by immunohistochemical methodThe expression of GR-? protein in the hormone resistance group was significantly higher than that in the normal group(P<0.01),while the other three groups showed no significant difference compared with the hormone resistance group(P>0.05).Compared with the normal group,the expression of GR-? protein in the hormone resistance group was significantly reduced(P<0.01);Compared with the hormone resistance group,there was no significant change in the expression of GR-? protein in the dexamethasone group(P<0.01),while it was significantly increased in Chaipu Decoction group and dexamethasone+Chaipu Decoction group(P<0.05,P<0.05),and there were no differences between Chaipu Decoction group and dexamethasone+Chaipu Decoction group.Compared with the normal group,the expression of AP-1 protein in the hormone resistance group increased significantly(P<0.05).Compared with the hormone resistance group,the expression of AP-1 protein in dexamethasone group and Chaipu Decoction group was significantly lower(P<0.05),and the difference between the two groups was not significant(P>0.05),while the decrease in the dexamethasone+Chaipu Decoction group was the most obvious(P<0.01).The expression of AP-1 protein in dexamethasone group and chaipu decoction group was significantly lower than that in hormone resistance group(P<0.05),but the differences between the two groups were not significant(P>0.05),while the decrease in the dexamethasone+Chaipu Decoction group was the most obvious(P<0.01).3.The expression of GR-?,GR-?,AP-1 mRNA in lung tissue by fluorescent quantitative PCR methodThe expression of GR-? mRNA in the hormone resistance group was significantly higher than that in the normal group(P<0.05),while the other three groups showed no significant difference compared with the hormone resistance group(P>0.05).Compared with the normal group,the expression of GR-? mRNA in the hormone resistance group was significantly reduced(P<0.01);Compared with the hormone resistance group,there was no significant change in the expression of GR-? mRNA in thedexamethasone group(P<0.01),while it was significantly increased in Chaipu Decoction group and dexamethasone+Chaipu Decoction group(P<0.05,P<0.05),and there were no differences between Chaipu Decoction group and dexamethasone+Chaipu Decoction group.Compared with the normal group,the expression of AP-1 mRNA in the hormone resistance group increased significantly(P<0.05),Compared with the hormone resistance group,the expression of AP-1 mRNA in dexamethasone group and Chaipu Decoction group was significantly lower(P<0.05),and the difference between the two groups was not significant(P>0.05),while the decrease in the dexamethasone+Chaipu Decoction group was the most obvious(P<0.01).The expression of AP-1 mRNA in dexamethasone group and chaipu decoction group was significantly lower than that in hormone resistance group(P<0.05),but the differences between the two groups were not significant(P> 0.05),while the decrease in the dexamethasone+Chaipu Decoction group was the most obvious(P<0.01).Conclusion: Chaipu Decoction could improve the therapeutic efficacies of glucocorticoid for steroid-resistant asthma and its mechanism might be through increasing the expression of GR-?,lowering the expression of AP-1?... |
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