The Effect Of Azithromycin On Expression Of Nuclear Factor-kappa Bp65(NF-κBp65) In Asthmatic Mice

Objective:To investigate the expression of nuclear factor in the blood and lung tissue of asthmatic mice and the effect of azithromycin on NF-κBp65 expression,as well as to analyze its role in asthma Pathogenesis.Methods:40 BALB/C were divided into four group.A is asthma model group.B is negative control group.C is azithromycin group.D is dexamethasone(Dex) group. ALL mice were sensitized with intraperitoneal ovalbumin plus alum on days 0 and 7 except B group.From day 14 to 21, the sensitized mice were challenged with an aerosol 1% ovalbumin daily for 30 minutes, and the asthma model was harvested on day 21.The C group and D group were seperately treated with azithromycin (50 mg/kg) and Dex (2 mg/kg) on days 14 to 21(1 h before the ovalbumin challenge). The B group were conducted by i.p injection of 0.5 ml NS on days 0 and 7, and challenged with NS daily for 30 minutes from day 14 to 21.Get the BALF to count the toal numember of the cell ,and the blood were used to test NF-κBp65 protein via ELISA.The lung were used to test the expression of NF-κBp65mRNA via RT-PCR, and the activity of NF-κBp65 in lung were tested via immunohistochemistry and western blot.Results:1. NF-κBp65mRNA expression between four groupsThe NF-κBp65mRNA expression in ACD groups was significantly increased as compared with the control group B. The NF-κBp65mRNA expression in CD groups was significantly decreased as compared with the group A, and the expression in D group is less than C group. 2. NF-κBp65 coloring degree via immunohistochemistry between four groupsThe coloring degree via Immunohistochemistry in ACD groups was significantly decreased as compared with the control group B. The coloring degree via Immunohistochemistry in CD groups was significantly increased as compared with the group A, and the degree in D group is more than C group.3. NF-κBp65 protein via western blot between four groupsThe NF-κBp65 protein via western blot in ACD groups was significantly increased as compared with the control group B. The NF-κBp65 protein via western blot in CD groups was significantly decreased as compared with the group A, and there were statistically significant between C and D group.4. NF-κBp65 protein in peripheral blood between four groupsThere were no statistically significant between four groups.5. Analyze the cell numember of the bronchoalveolar lavage fluid(BALF)The accumulation of total BALF immune cells in ACD groups was significantly increased compared with group B. Azithromycin treatment attenuated the accumulation of total BALF immune cells, compared with the group A. And the analysis of the BALF leukocyte populations demonstrated that azithromycin treatment significantly attenuated the accumulation of eosinophils and neutrophils, compared with group D.6. Corelation between NF-κBp65 protein in peripheral blood and NF-κBp65mRNA expression in lungThere was no correlation between NF-κBp65 protein in peripheral blood and NF-κBp65mRNA expression in lung.7. Corelation between the number of eosinophils in BALF and NF-κBp65mRNA expression in lungThe correlation between the number of eosinophils in BALF and NF-κBp65mRNA expression in lung is totally positive correlation.8. Corelation between the number of neutrophils in BALF and NF-κBp65mRNA expression in lung The correlation between the number of neutrophils in BALF and NF-κBp65mRNA expression in lung is totally positive correlation.Conclusion:1. The NF-κBp65 has played a role in the pathogenesis of asthma, and NF-κBp65mRNA expression can reflect both the number of eosinophils and neutrophils in the airway.2. There was no correlation between NF-κBp65 protein in peripheral blood and NF-κBp65mRNA expression in lung.So, NF-κBp65 protein in peripheral blood cannot reflect NF-κBp65mRNA expression in lung.3. Given the treatment with azithromycin can decrease the expression of the NF-κBp65mRNA in the lung of the asthmatic mice, that shows azithromycin can reduce the inflammation of airway by inhibited the activity of NF-κB. Compared with the given the treatment with DEX, the anti-inflammatory effect of azithromycin is less than DEX.