Background : Aquaporins (AQPs) may play a role in the pathogenesis of pulmonary inflammation and edema. This study investigated the role ofAQPs in acute lung injury following bleomycin inhalation in rats. Methods : Sprague-Dawley rats were treated via inhalation with 10 U/kg bleomycin hydrochloride dissolved in 5 ml of normal saline. The control rats were treated with 5 ml normal saline. The animals (n = 6-8 rats per group) were sacrificed at 4, 7, and 14 d. The changes in AQP1, AQP4, and AQP5 expression levels over time were analyzed by Western blotting. The nitrate and nitrite concentrations in the bronchoalveolar lavage fluid (BALF) were measured using a modified Griess reaction. ELISA was used to check cytokines. Results : The respiration rates were significantly higher 4 and 7 days after the bleomycin treatment compared with those of the control rats. The tidal volume was lower in rats at 4 days after the bleomycin treatment, and the wet/dry weights of the lung were significantly higher than those of the control group. The nitrite and nitrate concentrations in the BALF from the rats at 4 days after exposure to bleomycin were greater than those from the saline-treated rats. Immunoblotting studies demonstrated that the AQP1 and AQP4 expression levels were lower in the rats at 4 days. However, the AQP4 expression level was higher at 7 days. The AQP5 expression level increased at 4, 7 and 14 days after the bleomycin treatment. Conclusion : This study demonstrates that AQPs are expressed differently in bleomycin-induced pulmonary edema.
Jeon, Gang;Jang, Seung Hun;Song, Hae Geun;Ha, Jun-Wook;Eom, Kwang-Seok;Bahn, Joon-Woo;Kim, Dong-Gyu;Shin, Tae Rim;Park, Sang Myon;Park, Yong Bum;Kim, Chul-Hong;Hyun, In-Gyu;Jung, Ki-Suck
Tuberculosis and Respiratory Diseases
/
v.57
no.6
/
pp.535-542
/
2004
Background : Despite the clinical clues of bronchial asthma, some chronic coughers fail to be diagnosed due to negative test results. This study was aimed at evaluating the diagnostic performance of routine objective tests and identifying a cost-effective approach for asthmatics with a chronic cough. Methods : Patients with a chronic cough of more than 3 weeks duration, and showing normal chest radiograph and spirometry were enrolled. On the first visit, objective tests, composed of serum total IgE, peripheral blood eosinophil count, spontaneous sputum eosinophil count, methacholine bronchial provocation test (MBPT) and paranasal sinus radiograph, were performed, with the simultaneous administration of oral prednisolone (0.5mg/kg) for one week. The final diagnoses were made on the basis of the test results, and the patients grouped according to their steroid responsiveness. The role of the etiologic diagnosis tests was evaluated, and the medical costs of the final management plan simulated with respect to three assumed models. Results : Sixty chronic coughers were finally analyzed. The final diagnoses were as follows: bronchial asthma 21.7%, eosinophilic bronchitis 6.7%, paranasal sinusitis 18.3%, presumptive allergy 8.3% and non-diagnostic case 45.0%. Ninety percent were steroid responder. With the bronchial asthma cases, the positive rate of MBPT was 38.5%, with sputum eosinophil count in 84.6%, serum total IgE in 38.5%, and a peripheral blood eosinophil count rate of 30.8%. When the test results and steroid responsiveness data were applied to the 3 models, the chest radiograph, spirometry, sputum eosinophil count and paranasal sinus radiograph test results, and simultaneous short term steroid treatment seemed to have acceptable diagnostic performances, which could be used as a further guide to cost-effective planning. Conclusion : Objective tests, composed of chest radiograph, spirometry, paranasal sinus radiograph and sputum eosinophil count, with simultaneous short term steroid treatment, are suggested as cost-effective approaches for asthmatics with a chronic cough.
Kim, Hyung-Jung;Chae, Ho-Zoon;Ahn, Chul-Min;Kim, Sung-Kyu;Lee, Won-Young
Tuberculosis and Respiratory Diseases
/
v.47
no.4
/
pp.451-459
/
1999
Background : In sepsis, excessive generation of reactive oxygen species plays key roles in the pathogenesis of acute lung injury. The serum antioxidants such as catalase and MnSOD are elevated in sepsis and considered as predictors of acute respiratory distress syndrome(ARDS) and prognostic factors of sepsis. Peroxiredoxin(Prx) has recently been known as an unique and major intracellular antioxidant. In this study, we evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells(RAW 267.7) after treatment of oxidative stress and endotoxin and measured the amount of Prx I, Prx II and thioredoxin(Trx) in peritoneal and bronchoalveolar lavage fluid of septic animal model. Methods : Using immunoblot analysis with specific antibodies against Prx I, Prx II and Trx, we evaluated the distribution of Prx I and Prx II in human neutrophil, alveolar macrophage and red blood cell. We evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide(LPS) and measured the amount of Prx I, Prx II and Trx in peritoneal lavage fluid of intraperitoneal septic animals(septic animal model induced with intraperitoneal 6 mg/Kg LPS injection) and those in bronchoalveolar lavage fluid of intraperitoneal septic animals and intravenous septic animals(septic animal model induced with intravenous 5 mg/Kg LPS injection) and compared with the severity of lung inflammation. Results : The distribution of Prx I and Prx II were so different among human neutrophil, alveolar macrophage and red blood cell. The expression of Prx I in mouse monocyte-macrophage cells was increased after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide but that of Prx II was not increased. The amount of Prx I, Prx II and Trx were increased in peritoneal lavage fluid of intraperitoneal septic animals but were not increased in bronchoalveolar lavage fluid of intraperitoneal and intravenous septic animals regardless of the severity of lung inflammation. Conclusion : As intracellular antioxidant, the expression of Prx I is increased in mouse monocyte-macrophage cells after treatment of oxidative stress and endotoxin. The amount of Prx I, Prx II and Trx are increased in local inflammatory site but not increased in injured lung of septic animal model.
Background: To evaluate airway responses and inflammation to antigen in Sprague-Dawley rat asthma model, we examined airway responses, serial histologic changes of the lung, and the relationship between airway responses and airway inflammation after antigen airway challenge. Methods: Sprague-Dawley rats were sensitized with subcutaneous injection of 10 ${\mu}g$ ovalbumin(OA). Antigen airway challenges were done 14~16 days after sensitization and the sensitized rats were sacrificed 1h($A_E$), 6~8h($A_L$) and 1day($A_D$) after airway challenge, to examine the histologic changes of the lung. Airway responses were measured by body plethysmograph and recorded by enhanced pause(Penh) as an index of airway obstruction 6~8h after antigen challenges. Nonsensitized controls(10 rats) were also challenged with antigen and sacrificed 1 day later. Histopathologic examination of two trachea, large bronchi, small bronchi, and vessels was performed to evaluate the severity of inflammation and eosinophilic infiltration with H&E stain. Results: In 17 of 20 rats(85%) in both groups, we observed airway responses. Among them, an early response(ER) in 15 rats(75%), an dual response in 5(25%), and an late response(LR) only in 2 rats(10%) displayed. There were no significant differences in the severity of inflammation among the trachea, large bronchi, small bronchi and vessels in all groups after antigen challenge(p>0.05) and between early and late responders. The significant eosinophil infiltration was observed in 5 rats(50%) of AL(p<0.05) compared with in AE and controls. Also, eosinophil infiltration was observed in higher trend in LR(57.1%) compared to ER(40%)(p>0.05). Conclusion: Sprague-Dawley rats sensitized with subcutaneous injection of OA showed a significant airway responses to antigen challenge. But antigen challenges caused a little eosinophil infiltration and no significant airway inflammation. Asthma model of Sprague-Dawley rats could be useful for antigen-induced airway responses, but this model has a limitation for the study of human asthma because of no significant pathologic change.
Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.
Background : Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method : M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and $IFN-{\gamma}$ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result : Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and $IFN-{\gamma}$ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion : Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.
Background : Cyclooxygenase is the main target enzyme for the nonsteroidal anti inflammatory drugs (NSAIDs) that have been shown to suppress carcinogenesis in both experimental models and epidemiological studies. COX-2 plays an important role in solid tumor growth, invasiveness and angiogenesis, through, in part, the synthesis of prostaglandins, such as prostaglandin E2 (PGE2). In this study, the prognostic significance of an increase in COX-2 expression in lung cancer samples was evaluated. Material and Methods : The expression of COX-2, by immunohistochemistry, was studied in paraffin-embedded tumor blocks obtained from 84 patients(male 67, female 17, with a mean age of 63, ranging from 34 to 84 years) who had undergone surgery at Wonkwang University Hospital, between 1997 and 2002. For the evaluation of the relationships between COX-2 expression, and the clinical stage, metastasis to lymph nodes and survival, those cases showing the respective antigen expression in >10% of the tumor cells were considered positive. Result : Of the 84 patients, 61 (73%) exhibited more than 10% COX-2 immunoreactivities in the tumor and normal cells, whereas the remaining 23 showed no increase in the expression of COX-2. There was no significant relationship between the increased expression of COX-2 and the disease stage(p=0.1002) or cell type(p=0.152). The median survival was longer for the patients with a negative, compared to positive, COX-2 expression(36 compared to 24 months, p<0.05). The two year-survival rate was also higher in the patients with a negative COX-2 expression (78%) than those with a positive expression (47%, Kaplan-Meier, Log Rank, p < 0.05). Conclusion : The median survival was longer in the patients with a negative, compared to positive, COX-2 expression was longer than those with positive COX-2, having undergone complete resection due to primary non-small cell lung cancer.
The respiratory tract is an essential part of the respiratory system involved in the process of respiration. However, if stenosis occurs, it interferes with breathing and can even lead to death. Asthma is a typical example of a reversible cause of airway narrowing, and the number of patients suffering from acute exacerbation is steadily increasing. Therefore, it is important to detect airway narrowing early and prevent the patient's condition from worsening. Optical coherence tomography (OCT), which has high resolution, is suitable for observing the microstructure of tissues. In this study we developed an endoscopic OCT system. We combined a 1300-nm OCT system with a servo motor, which can rotate at a high speed. A catheter was pulled back using a linear stage while imaging with 360° rotation by the motor. The motor was selected considering various requirements, such as torque, rotational speed, and gear ratio of pulleys. An ex vivo rabbit tracheal model was used as a sample, and the sample and catheter were immobilized by acrylic structures. The OCT images provided information about the structures of the mucosa and submucosa. The difference between normal and stenosed parts in the trachea was confirmed by OCT. Furthermore, through a three-dimensional (3-D) reconstruction process, it was possible to identify and diagnose the stenosis in the 3-D image of the airway, as well as the cross-sectional image. This study would be useful not only for diagnosing airway stenosis, but also for realizing 3-D imaging.
Lee, Jae A;Jin, Gong Yong;Bok, Se Mi;Han, Young Min;Park, Seoung Ju;Lee, Yong Chul;Chung, Myung Ja;Youn, Gun Ha
Tuberculosis and Respiratory Diseases
/
v.67
no.5
/
pp.436-444
/
2009
Background: Micro computed tomography (CT) is rapidly developing as an imaging tool, especially for mice, which have become the experimental animal of choice for many pulmonary disease studies. We evaluated the usefulness of micro CT for evaluating lung fibrosis in the murine model of bleomycin-induced lung inflammation and fibrosis. Methods: The control mice (n=10) were treated with saline. The murine model of lung fibrosis (n=60) was established by administering bleomycin intra-tracheally. Among the 70 mice, only 20 mice had successful imaging analyses. We analyzed the micro CT and pathological findings and examined the correlation between imaging scoring in micro CT and histological scoring of pulmonary inflammation or fibrosis. Results: The control group showed normal findings on micro CT. The abnormal findings on micro CT performed at 3 weeks after the administration of bleomycin were ground-glass opacity (GGO) and consolidation. At 6 weeks after bleomycin administration, micro CT showed various patterns such as GGO, consolidation, bronchiectasis, small nodules, and reticular opacity. GGO (r=0.84) and consolidation (r=0.69) on micro CT were significantly correlated with histological scoring that reflected pulmonary inflammation (p<0.05). In addition, bronchiectasis (r=0.63) and reticular opacity (r=0.83) on micro CT shown at 6 weeks after bleomycin administration correlated with histological scoring that reflected lung fibrosis (p<0.05). Conclusion: These results suggest that micro CT findings from a murine model of bleomycin-induced lung fibrosis reflect pathologic findings, and micro CT may be useful for predicting bleomycin-induced lung inflammation and fibrosis in mice.
Kim, Soo Hyun;Kim, Min Ju;Shin, Mi-Rae;Roh, Seong-Soo;Kim, Seung Hyung;Park, Hae-Jin
The Korea Journal of Herbology
/
v.37
no.3
/
pp.29-39
/
2022
Objective : This study is aimed to evaluate the protective effects of Rehmanniae Radix, Mori Folium, and Liriopie Tuber mixture (RML) on lung injury of Particulate matter less than 10 um in diameter and diesel exhaust particles (PM10D) mice model. Methods : To investigate the anti-inflammatory activity of RML, PM10D was diluted in aluminum hydroxide (Alum) in 7-week-old male mice and induced by Intra-Nazal-Tracheal (INT) injection method. Animal experiments were divided into 5 groups. Nor (normal mice), CTL (PM10D-induced mice with the administration of distilled water), DEXA (PM10D-induced mice with the administration of 3 mg/kg Dexamethasone), RML 100 (PM10D-induced mice treated with RML 100 mg/kg weight), and RML 200 (PM10D-induced mice treated with RML 200 mg/kg body weight). After 11 days administration, mice were sacrificed and inflammation-related immune cells in broncho-alveolar lavage fluid (BALF) were analyzed. Inflammation-related biomarkers were also analyzed in blood and lungs. Lung tissue was observed through histological examination. Results : In the PM10D induced model, the PML showed decreases in CXCL-1 and IL-17A in BALF. Expression of inflammatory cytokines and cough-related mRNA genes was significantly decreased in serum and lung tissue. The mixture treatment of RML significantly improved the immune related cells in the serum. In addition, histological observations showed a tendency to decrease the severity of lung injury. Conclusions : Overall, these results confirmed the respiratory protective effect of the RML mixture in a model of lung injury induced by air pollution (PM10+DEP), suggesting that it is a potential treatment for respiratory damage.
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