Alteration of apoptotic susceptibility and bcl-2 gene in malignant transformation of human bronchial epithelial cells

AIM: To demonstrate the susceptibility of cell apoptosis varies during the progress of cell malignant transformation from human being in vitro. METHODS: A SV₄₀T-transfected human bronchial epithelial immortalized cell line (called M) was selected in this work, which has acquired some characteristics of malignant transformation at the later passage. The alterations of apoptosis and bcl-2, P53 genes between early and later passage of M cells were investigated by means of TDT labeling in situ, chromosome FISH, RNA and protein testing, etc. RESULTS: Incidence of apoptosis induced by cis-platin was significantly lower in later than in early passages of M. Levels of bcl-2 mRNA and protein in later passages were higher than early passages of M, and overexpression of bcl-2 was accumulated following the development of cellular malignancy. P53 protein level was as high in early as in later passages. CONCLUSION: Overexpression of bcl-2 decreases the cellular sensitivity to apoptotic inductors plays an important role during progress of carcinogenesis in human bronchial epithelial cancers. The inactivation of P53 protein in the SV₄₀-T transfected M cell line may be one of reasons of bcl-2 overexpression, but not associated with the accumulation of bcl-2 expressed level during cell transformation.     

Oxidative Stress and Interaction of Endothelin Receptors in Airways of Clinically Healthy Horses

Endothelin-1 (ET-1), a physiological as well as an inflammatory mediator, causes contraction of airway smooth muscles by binding to endothelin-A (ETA) and endothelin-B (ETB) receptors. We investigated the interaction between endothelin receptors on contractions and oxidative stress of bronchial rings of clinically healthy horses. Rings for response studies were set in tissue chambers containing oxygenated Tyrode’s solution. Concentration-response relationships were determined for ET-1 on control rings, rings incubated with ETA antagonist (BQ123), rings incubated with ETB antagonists (IRL 1038 and BQ788), and rings incubated with all antagonists combined. For oxidative stress studies, rings were incubated for 30 minutes separately with ET-1, ETA antagonist + ET-1, ETB antagonists + ET-1, and all three antagonists + ET-1. The control rings were not treated with any agents. Electron paramagnetic resonance (EPR) method was used to measure total reactive oxygen species (ROS), superoxide, and peroxynitrite. Results showed that ET-1 caused dose-dependent contractions which were increased by ETA blockade and decreased by ETB blockade. Combined blockade significantly inhibited the response to ET-1. Regarding the oxidative stress, ET-1 and its antagonists significantly reduced the total ROS. Superoxide production was significantly decreased by ETA antagonist and completely abolished when ETA and ETB antagonists were combined. Peroxynitrite production was not changed significantly. The study suggested that an interaction between ET receptors exists in the expression of contractile responses and oxidative stress. ET-1 attenuated oxidative stress by decreasing total ROS. ETA receptors may be primarily responsible for superoxide production. It appears that ET-1 does not affect peroxynitrite production in normal equine tissues.     

Effects of ginsenoside Rh1 on expression of inflammatory factors in mouse asthma model

AIM: To observe the effects of ginsenoside Rh1 on the levels of inflammatory factors in serum and bronchoalveolar lavage fluid (BALF), and the pathological changes of the lung tissues in an experimentally induced mouse asthma model. METHODS: Male BALB/c mice (n=40) were divided into 4 groups:normal control group, asthma mo-del group, and low-dose (40 mg·kg^-1·d^-1) and high-dose (80 mg·kg^-1·d^-1) ginsenoside Rh1 groups. The bronchial asthma mouse model was established by the method of ovalbumin induction and excitation, and during the excitation period, the mice were daily treated with ginsenoside Rh1 for 2 weeks. At 24 h after the final dose of ginsenoside Rh1, the mice were sacrificed. The number of eosinophils (EOS) and the concentrations of interleukin (IL)-4, IL-5 and interferon (IFN)-γ in BALF were determined. The levels of IgG and IgE in serum were measured, and the expression of transforming growth factor (TGF)-β1 and the pathological changes in lung tissues were evaluated. RESULTS: Ginsenoside Rh1 inhibited the increases in the number of EOS and the concentrations of IL-4, IL-5, IFN-γ and IgE, reversed the increased expression of TGF-β1, and improved the pathological changes of the lung tissues in asthmatic mice. CONCLUSION: Ginsenoside Rh1 improves the immuno-inflammatory profile and pathological changes in the experimentally induced mouse asthma model, implying its potential therapeutic effect on asthma.     

慢性持续期支气管哮喘患者痰液中IL-4水平与其临床基线评估相关性分析

目的:探讨慢性持续期支气管哮喘患者痰液中白细胞介素-4(IL-4)水平与其临床基线评估的相关性。方法:分别对慢性持续期支气管哮喘患者轻度16例、中度14例、重度18例进行临床基线评估,主要是肺功能检测及哮喘症状控制评分;并同时采用酶联免疫吸附试验(ELISA)法检测其诱导痰液中IL-4水平。结果:慢性持续期支气管哮喘患者痰液中IL-4水平与肺通气功能指标FEV1%呈负相关,与哮喘发作程度临床症状评分相一致。结论:慢性持续期支气管哮喘患者痰液中IL-4水平与其发作程度有关,能作为哮喘评估的一项指标。     

三黄鸡“支气管”堵塞病的诊断和治疗

鸡的许多传染病都会造成呼吸系统损伤,并可导致大批死亡。2010年后,新疆乌鲁木齐周边地区的一些鸡群都出现了一种以气管出血、气囊浑浊和支气管被淡黄色干酪样物质堵塞为主要病理变化的呼吸系统疫病,其发病率和死亡率高,危害严重。本文对一例典型“支气管”堵塞病例,采集病料后制作石蜡包埋切片,进行了病理组织学检查,并用中药“法贝灵”为主的药物组方进行了鸡群治疗。病理组织学观察发现,患鸡的初级支气管、次级支气管和三级支气管均被炎性渗出物堵塞;“法贝灵”为主的药物组方治疗效果显著。     

Deciphering the Role of Bronchial Hyper-Responsiveness in Equine Pasture Asthma

Bronchial hyper-responsiveness (BHR) describes a lung abnormality in which airways are easily triggered to constrict in response to normally harmless inhaled stimuli, and is a key element of human asthma pathophysiology. BHR contributes to equine respiratory diseases including inflammatory airway disease and recurrent airway obstruction. Collectively these diseases account for over 80% of poor performance in equine athletes, and at least 10% of veterinary admissions. BHR is also a contributing factor in ‘exercise induced pulmonary hemorrhage’. Increased sensitivity to airway constriction that characterizes BHR is a documented sequel to viral respiratory infections in several species, including horses and humans. Five respiratory viruses known to circulate extensively in equine populations place the horse at risk for BHR. Despite adverse effects of BHR on equine health, there remains a gap in our fundamental understanding of how gene products coordinate in the lung to cause BHR. Leveraging the equine genome sequence, we employ systems biology including proteomics and RNA sequencing to model the complex biology of BHR in the lungs of horses with pasture asthma. Using a self controlled experimental design, gene products that segregate with seasonal asthma exacerbation in diseased horses are being identified and their relevant physiology identified to address the need for better recognition and management of BHR in equine disease.     

Effects of damage on endothelin production in epithelial cells and sub-epithelial fibroblasts

AIM: To investigate the level of ET-1 produced by cultured human bronchial epithelial cells (HBECs) under injury and the effects of injured HBECs on ET-1 production in sub-epithelial fibroblasts. The interaction between ET-1 and matrix metalloproteinase-9(MMP-9) was detected in HBECs under damage. The purpose of the study is to evaluate the effect of injured HBECs related to ET-1 release on airway remodeling in asthma. METHODS: ET-1 level was detected in supernatants from cultured HBECs 12 h after being treated with either mechanical scraping or LPS stimulation or mechanical scraping plus LPS, as well as from subepithelial fibroblasts cocultured with mechanical damaged HBECs. It was also measured in the supernatant from HBECs transfected with MMP-9 expression plasmid. MMP-9 activity was assessed in supernatants from HBECs stably transfected with pEGFPc1 -antisense-ET-1 converting enzyme(ECE) RNA. RESULTS: It was found that there was an increase in ET-1 level in supernatants from HBECs either treated with mechanical scraping plus LPS or transiently transfected with MMP-9 plasmid, as well as from sub-epithelial fibroblasts cocultured with mechanical scraping HBECs compared with that in controls. Gelatin zymography showed a obviously attenuated gelatinolytic activity of MMP-9 in conditioned media of HBECs expressing antisense ECE RNA after mechanical damage. CONCLUSIONS: Airway epithelial cells under injury are able to overproduce ET-1 as well as initiate ET-1 release from sub-epithelial fibroblasts, MMP-9 produced by injured bronchial epithelial cells may also increase ET-1 processing leading to ET-1 production further. The interaction between ET-1 and MMP-9, both of which enhanced in damaged HBECs, may play an important role in airway inflammation related to airway remodeling in asthma.     

Eosinophilic bronchopneumopathy in dogs

Eosinophilic bronchopneumopathy was diagnosed in 23 young dogs. Clinical signs included cough, gagging, and retching in all dogs, dyspnea in 21 dogs (91%), and nasal discharge in 12 dogs (52%). The most common radiographic findings were a moderate to severe bronchointerstitial pattern (68%, 13 of 19 dogs). Bronchoscopic findings included the presence of abundant yellow-green mucus or mucopurulent material (70%, 16 of 23 dogs) and severe mucosal thickening with an irregular or polypoid appearance (52%, 12 of 23 dogs), with partial airway closure during expiration in 3 dogs (13%). Peripheral blood eosinophilia was noted in 14 of 23 dogs (61%). Inflammatory cells in brush or bronchoalveolar lavage fluid cytologic preparations comprised more than 50% eosinophils in 14 of 23 dogs (61%), and 20-50% eosinophils in 6 dogs (26%). Eosinophilic infiltration of the bronchial mucosa was observed in biopsies from 19 dogs and was graded as mild (37%, 7 dogs), moderate (32%, 6 dogs), or severe (32%, 6 dogs). The mean serum immunoglobulin A concentration was almost double that of a population of 20 healthy dogs of various breeds. Oral glucocorticoids were administered on alternate days with progressive tapering of the dose; the dosage at maintenance varied between 0.1 and 1.0 mg/kg every other day. No relationship was found between the duration of clinical signs and the maintenance dosage or the cytologic and histopathologic grades.