| Ojective:As a one of anticholinergic drug, Ipratropium bromide can inhibit the parasympathetic nerve fibers in the nasal cavity, clinically; it used to control nasal symptoms of allergic rhinitis. It’s rarely reported at home and abroad about this respect that whether Ipratropium bromide can have an impact on the body’s cellular and humoral immunity with relieving the symptoms of allergic rhinitis. Through the construction of AR animal models, the experiment is to investigate the effects of anticholinergic drugs on intranasal immunization environment of AR animal models, and to provide a theoretical basis for the clinical treatment of anti-cholinergic drugs.Methods:32female BALB/c mice with5-6week old were randomly divided into two groups:16of the mice were used to establish a model of allergic rhinitis, and another16for the controls. Sensitization Methods: Using the classical OVA sensitization method to construct the AR model. The method was that the mice of the experimental group were immunized by an intraperitoneal injection on day0and day5, then hired OVA from the day12for7consecutive days of intranasal challenge, and then used OVA once more slowly for nasal stimulation every other day for10weeks. The rest of the groups used saline instead. In order to simulate the chronic process of allergic rhinitis the entire modeling process spent more than4months, After11mice succeed, they were randomly divided into model group (5mice) and treatment model group (6mice), the control group was also divided into blank control group (8) and the treatment control group (8). The mice of treatment model group and treatment control group began to be treated with3mg/ml ipratropium20μl in each side of the nasal for nasal intranasal, and continuously administered for4week, at the same time, the model group and the control group are used saline instead of ipratropium bromide intranasal. During the treatment, the model group and the treatment model group continued to be given3% OVA solution10μl in each side of the nasal for nasal stimulation. After treatment, take some nasal mucosa of mice, then comparing the tissue morphology by observing HE staining. Due to the mice died and other reasons, the numbers of final samples for analysis were5mice of the control group,3of model group,5of the treatment model group, and5of the treatment control group. Finally, detected the IL-4, IL-17, IFN-y, Foxp3protein content of each group of mice by Western blot method, and detected contents of IL-4, IL-17mRNA with RT-PCR method.Results:Before the treatment, the behavior scores of the treatment model group and the model group were reached or exceeded allergic rhinitis standard of scores, they showed that construction of model were successful. After the treatment, the behavior scores of the treatment model group was lower than the model group (P<0.05).The results of Western blot showed that compared with the control group, in the model of allergic mice, nasal express IL-4and IL-17were significantly higher (P<0.05), IFN-y and Foxp3protein expression were also slightly elevated, but they were not statistical significances (P>0.05). After the Ipratropium bromide treatment for allergic rhinitis topically, IL-4expression was significantly decreased in allergic mice (P<0.05), whereas Foxp3expression tended to increase, but they were not statistical differences (p>0.05), besides, IFN-y and IL-17cytokine expression was not significant changes (p>0.05). After Ipratropium bromide treatment for control groups, nasal topical IL-4, IL-17, Foxp3expression tended to increase.The results of RT-PCR showed that compared with the control group, expression of IL-4, IL-17mRNA have increased in allergic rhinitis mice (p<0.05). After Ipratropium bromide treatment, IL-4expression have a downward trend (p<0.05), the results were the same as the Western blot, and IL17-expression was also significantly decreased (p<0.05).Conclusion:1) Through the analysis of Western blot and RT-PCR, there had immune imbalance which based advantages of Th2cells in the nasal mucosa of allergic mice. At the same time, IL-17expression was also significantly increased.2) Ipratropium bromide, while improving the symptoms of allergic rhinitis, can reduced cytokines IL-4 which secreted by Th2cells of nasal mucosa, and enhanced the generation of Foxp3Treg cells, but did not change the expression of IL-17and IFN-y.3) Long-term use of Ipratropium bromide may have a certain degree of inflammation to nasal mucosa of normal mice, such as cytokines IL-4and IL-17, Foxp3expression showed elevated reaction.9... |
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