Research Tiotropium On Lung Surface -D Chronic Obstructive Pulmonary Disease Activity Of The Protein

Objective:To established the COPD rat model by passive smoking and intratracheal instillation of lipopolysaccharide (LPS).Methods:Twenty SPF healthy male SD rats,which were6-8weeks of age and the initial weight were (200±20) g.They were randomly divided into a control group and a COPD group (n=10in each group). The COPD group rats were established by intratracheal instillation of LPS at the30th ang60th day.The rest of the day they were smoked by5%smoke concentration twice daily and30minutes once.The intervals between two times is8hours. Total length is90days.After the model established, we took the lung tissue to research.The pathologic change of the lung tissue and airway were observed by HE staining. Emphysema changes were evaluated by mean linear intercept(MLI) of lung and mean alveolar number(MAN). The level of IN-8、TNF-α in serum and lung tissue were measured by enzyme-linked immunosorbent assay (ELISA).Results:After the model established, the COPD group rats were significant reduction in weight gaining and cough, short of breah and so on. HE staining showed significant lumen distortion, lodging of cilia, exuviations, increase of secretion in lumens, alveolar cavity expanding into pulmonary bullae, infiltration of inflammatory cells. The COPD group appears obvious emphysema.The expression of IN-8、TNF-α in serum and lung tissue of the COPD group were obviously stronger than that in the control group (P<0.01). All of these proved that the rat COPD models were successfully established.Conclusion:The method of passive smoking and intratracheal instillation of LPS could successfuly make the rat model of COPD,which could be used in experimental study. Objective:To investigate the new mechanism of tiotropium bromide in the treatment of chronic obstructive pulmonary disease (COPD) by measuring the tiotropium bromide-caused changes in the levels of Pulmonary Surfactant Protein D (SP-D) in the serum and lung tissues before and after establishment of the COPD rat model.Methods:Thirty SPF healthy male SD rats were provided by experimental animal center of ZheJiang province,which were6-8weeks of age and the initial weight were (200±20) g.They were randomly divided into a control group, a COPD group and a tiotropium bromide treatment group (n=10in each group). The COPD group and tiotropium bromide treatment group rats were established by intratracheal instillation of LPS at the30th ang60th day.The rest of the day they were smoked by5%smoke concentration twice daily and30minutes once.The intervals between two times is8hours. Total length is90days.The tiotropium bromide treatment group rats atomizing inhaled tiotropium bromide before Fumed by Cigarettes. After the model established, we took the lung tissue to research.The pathologic change of the lung tissue and airway were observed by HE staining. Emphysema changes were evaluated by mean linear intercept(MLI) of lung and mean alveolar number(MAN). The level of SP-D in serum was measured by enzyme-linked immunosorbent assay (ELISA).The expression of SP-DmRNA were tested by RT-PCR.The level of SP-D in lung tissue was detected by western-blot and immunohistochemistry.Results: The COPD group appears obvious emphysema, while the treatment group appears mild emphysema. The expression of SP-D in serum of the control,the COPD group and the treatment group were (42.14+2.52) ng/ml,(53.21+4.17) ng/ml and (47.58+5.97) ng/ml respectively (P<0.01).The expression of SP-DmRNA in lung tissue of the control,the COPD group and the treatment group were0.050±0.004,0.335±0.032and0.231±0.018(P<0.01).The level of SP-D in lung tissue of the three groups were0.68+0.01,0.98+0.01and0.76+0.01.Conclusion:1、The level of SP-D in serum and lung tissue of COPD were closely related to the mechanism of COPD.2、Tiotropium bromide could reduce the level of SP-D in serum and lung tissue of COPD rat.