In this study, the effect of prophylactic anti inflammation on the development of smoke induced emphysema was investigated. Young male guinea pigs aged 1.5 - 2 months (weighing 198.3±26.9 g) were randomly divi...In this study, the effect of prophylactic anti inflammation on the development of smoke induced emphysema was investigated. Young male guinea pigs aged 1.5 - 2 months (weighing 198.3±26.9 g) were randomly divided into 4 groups: group A (cigarette smoke exposure only), group B (cigarette smoke exposure plus pentoxifylline rich (PTX, 10 mg/d) forage feeding), group C (cigarette smoke exposure plus intermittent cortical steroid injection (Triamcinolone acetonide, 3 mg, im, every three weeks) and control group (group D: animals with sham smoke exposure, raised under the same conditions). Animals in group A, B and C were exposed to smoke of cigarettes for 1 to 1.5 h twice a day, 5 days a week. All animals were killed at the 16th week and followed by morphometrical analysis of the midsagittal sectioned lung slices. Smoke exposure of 16 weeks resulted in visible emphysematous development in Group A but not in Group B and C. It was evidenced by the indicator of air space size, mean linear intercept (L m): 120.6±16.0 μm in Group A; 89.8±9.2 μm in Group B and 102.4±17.7 μm in Group C. The average L m in either group B or group C was shorter than that in Group A (ANOVA and Newman Keuls test, F=8.80, P =0.0002) but comparable to that (94.8±13.2 μm) in group D ( P >0.05). It is concluded that long term prophylactic anti inflammation inhibits pulmonary emphysema induced by cigarette smoking in the guinea pigs.展开更多
Background Chronic obstructive pulmonary disease(COPD)is a severe public health problem.Cigarette smoke(CS)is a risk factor for COPD and lung cancer.The underlying molecular mechanisms of CS-induced malignant transfor...Background Chronic obstructive pulmonary disease(COPD)is a severe public health problem.Cigarette smoke(CS)is a risk factor for COPD and lung cancer.The underlying molecular mechanisms of CS-induced malignant transformation of bronchial epithelial cells remain unclear.In this study,we describe a lung-on-a-chip to explore the possible mechanistic link between cigarette smoke extract(CSE)-associated COPD and lung cancer.Methods An in vitro lung-on-a-chip model was used to simulate pulmonary epithelial cells and vascular endothelial cells with CSE.The levels of IL-6 and TNF-αwere tested as indicators of inflammation using an enzyme-linked immune sorbent assay.Apical junction complex mRNA expression was detected with qRT-PCR as the index of epithelial-to-mesenchymal transition(EMT).The effects of CSE on the phosphorylation of signal transduction and transcriptional activator 3(STAT3)were detected by Western blotting.Flow cytometry was performed to investigate the effects of this proto-oncogene on cell cycle distribution.Results Inflammation caused by CSE was achieved in a lung-on-a-chip model with a mimetic movement.CSE exposure induced the degradation of intercellular connections and triggered the EMT process.CSE exposure also activated the phosphorylation of proto-oncogene STAT3,while these effects were inhibited with HJC0152.Conclusions CSE exposure in the lung-on-a-chip model caused activation of STAT3 in epithelial cells and endothelial cells.HJC0152,an inhibitor of activated STAT3,could be a potential treatment for CS-associated COPD and lung cancer.展开更多
Background Cigarette smoke-induced emphysema is associated with overexpression of the chemokine receptor CXCR3 and its ligands. Previously, we have demonstrated that pentoxifylline （PTX） alleviated cigarette smoke-i...Background Cigarette smoke-induced emphysema is associated with overexpression of the chemokine receptor CXCR3 and its ligands. Previously, we have demonstrated that pentoxifylline （PTX） alleviated cigarette smoke-induced emphysema. The aim of this study was to determine if the overexpression of CXCR3 and its ligand interferon-inducible protein-10 （IP-10） that was elicited by smoke exposure were attenuated by PTX. Methods （1） The study in vitro： a given number of RAW264.7 macrophages with decreasing concentrations of PTX in the culture medium were challenged with cigarette smoke extract （CSE）; （2） The study in vivo： male BALB/c mice were randomized into four groups, i.e., sham-smoke, smoke only, smoke with 2 mg/kg PTX, and smoke with 10 mg/kg PTX. The smoke exposure time was 90 minutes once a day, 6 days a week for 16 weeks. PTX was given intraperitoneally before each episode of smoke exposure. Interferon （IFN）-y and IP-10 in broncho-alveolar lavage fluid （BALF） and in culture medium were measured by enzyme-linked immunosorbent assay （ELISA）. IP-10 mRNA in lung tissue was assessed by RT-PCR. CXCR3 positive cells in lung sections were visualized by immunochemistry staining. Results Up-regulation of IFN-γ and IP-10 in the culture medium of macrophages elicited by CSE was inhibited by PTX in a dose-dependent manner. Chronic cigarette smoke exposure led to overexpression of IFN-γ and IP-10 in BALF, upregulation of IP-10 mRNA and increased infiltration of CXCR3^＋ cells into lung parenchyma. Administration of PTX decreased the level of IFN-y from （6.26±1.38） ng/ml to （4.43±0.66） ng/ml by low dose PTX or to （1.74±0.28） ng/ml by high dose PTX. IP-10 was reduced from （10.35±1.49） ng/ml to （8.19±0.79） ng/ml by low dose PTX or to （7.51±0.60） ng/ml by high dose PTX. The expression of IP-10 mRNA was also down-regulated （P 〈0.05）. But only with a high dose of PTX was the ratio of CXCR3^＋ cells decreased; 15.2±7.3 vs. 10.4±1.8 （P 〈0.05）. Conclusion PTX attenuates cigarette smoke-induced overexpression of chemokine receptor CXCR3 and its ligand IP-10, which is relevant to its inhibitory effect on pulmonary emphysema.展开更多
目的探讨半胱氨酸天冬氨酸蛋白酶3(cysteinyl aspartate specific proteinase-3,Caspase-3)/Gasdermin-E(GSDME)介导的细胞焦亡在烟草烟雾(CS)暴露诱导的骨骼肌萎缩中的作用。方法将C57BL/6小鼠暴露于烟草烟雾24周,构建肺气肿小鼠模型...目的探讨半胱氨酸天冬氨酸蛋白酶3(cysteinyl aspartate specific proteinase-3,Caspase-3)/Gasdermin-E(GSDME)介导的细胞焦亡在烟草烟雾(CS)暴露诱导的骨骼肌萎缩中的作用。方法将C57BL/6小鼠暴露于烟草烟雾24周,构建肺气肿小鼠模型。HE染色观察腓肠肌组织的形态学变化;免疫组化检测腓肠肌中焦亡相关蛋白表达。用烟草烟雾提取物(CSE)处理C2C12小鼠骨骼肌细胞,建立体外骨骼肌萎缩模型。进一步采用Caspase-3抑制剂Z-DEVD-FMK和GSDME抑制剂Dimethyl fumarate(DMF)处理C2C12细胞。观察抑制Caspase-3/GSDME后对CSE诱导的骨骼肌萎缩的影响。用肿瘤坏死因子α(TNF-α)刺激骨骼肌细胞,观察TNF-α对Caspase-3/GSDME蛋白表达和肌管的影响。Western blot用于测定Caspase-3和GSDME蛋白的表达水平;Hoechst33342/Propidium Iodide(PI)双染色检测PI阳性细胞率;乳酸脱氢酶(LDH)释放实验检测C2C12细胞的LDH释放量;免疫荧光检测C2C12肌管直径变化。结果CS诱导小鼠腓肠肌萎缩的同时伴随着GSDME介导的焦亡增强(P<0.05)。体外实验结果表明,与对照组相比,CSE组C2C12细胞cleaved-caspase-3、GSDME-N的蛋白表达显著增高,PI阳性率和LDH释放率增加,肌管直径减小(P<0.05)。分别抑制Caspase-3和GSDME均可明显改善CSE诱导的骨骼肌细胞焦亡和肌管萎缩(P<0.05)。TNF-α促进肌管萎缩和骨骼肌细胞cleaved-caspase-3、GSDME-N蛋白的表达。结论CS可以通过TNF-α激活caspase-3/GSDME通路介导的焦亡,促进骨骼肌萎缩。展开更多
文摘In this study, the effect of prophylactic anti inflammation on the development of smoke induced emphysema was investigated. Young male guinea pigs aged 1.5 - 2 months (weighing 198.3±26.9 g) were randomly divided into 4 groups: group A (cigarette smoke exposure only), group B (cigarette smoke exposure plus pentoxifylline rich (PTX, 10 mg/d) forage feeding), group C (cigarette smoke exposure plus intermittent cortical steroid injection (Triamcinolone acetonide, 3 mg, im, every three weeks) and control group (group D: animals with sham smoke exposure, raised under the same conditions). Animals in group A, B and C were exposed to smoke of cigarettes for 1 to 1.5 h twice a day, 5 days a week. All animals were killed at the 16th week and followed by morphometrical analysis of the midsagittal sectioned lung slices. Smoke exposure of 16 weeks resulted in visible emphysematous development in Group A but not in Group B and C. It was evidenced by the indicator of air space size, mean linear intercept (L m): 120.6±16.0 μm in Group A; 89.8±9.2 μm in Group B and 102.4±17.7 μm in Group C. The average L m in either group B or group C was shorter than that in Group A (ANOVA and Newman Keuls test, F=8.80, P =0.0002) but comparable to that (94.8±13.2 μm) in group D ( P >0.05). It is concluded that long term prophylactic anti inflammation inhibits pulmonary emphysema induced by cigarette smoking in the guinea pigs.
基金This work was supported by National Natural Science Foundation of China(Grant Nos.81672297,Grant Nos.61701493)Policy Guidance project(International Science and Technology Cooperation)of Jiangsu Province of China(BZ2018040)+3 种基金the Natural Science Foundation of Tianjin,P.R.China(18JCYBJC42100)Hundred Talents Program of the Chinese Academy of SciencesProject Funded by China Postdoctoral Science Foundation(2019M651959)Postdoctoral Research Funding Program of Jiangsu Province(2018K004B).
文摘Background Chronic obstructive pulmonary disease(COPD)is a severe public health problem.Cigarette smoke(CS)is a risk factor for COPD and lung cancer.The underlying molecular mechanisms of CS-induced malignant transformation of bronchial epithelial cells remain unclear.In this study,we describe a lung-on-a-chip to explore the possible mechanistic link between cigarette smoke extract(CSE)-associated COPD and lung cancer.Methods An in vitro lung-on-a-chip model was used to simulate pulmonary epithelial cells and vascular endothelial cells with CSE.The levels of IL-6 and TNF-αwere tested as indicators of inflammation using an enzyme-linked immune sorbent assay.Apical junction complex mRNA expression was detected with qRT-PCR as the index of epithelial-to-mesenchymal transition(EMT).The effects of CSE on the phosphorylation of signal transduction and transcriptional activator 3(STAT3)were detected by Western blotting.Flow cytometry was performed to investigate the effects of this proto-oncogene on cell cycle distribution.Results Inflammation caused by CSE was achieved in a lung-on-a-chip model with a mimetic movement.CSE exposure induced the degradation of intercellular connections and triggered the EMT process.CSE exposure also activated the phosphorylation of proto-oncogene STAT3,while these effects were inhibited with HJC0152.Conclusions CSE exposure in the lung-on-a-chip model caused activation of STAT3 in epithelial cells and endothelial cells.HJC0152,an inhibitor of activated STAT3,could be a potential treatment for CS-associated COPD and lung cancer.
基金This study was supported by a grant from the Natural Science Foundation of Hubei Province, China （No. 2008cdb 153）. The authors have no conflict of interest to declare.Acknowledgements： We are grateful to Dr. TIAN Yuan, Surgery Laboratory of Union Hospital, Tongji Medical College for his generous donation of the macrophage cell line and guidance on cell culture. We also thank Prof. WU Ping and Ms. O1oo Stella Anne for their assistance on revision of this manuscript.
文摘Background Cigarette smoke-induced emphysema is associated with overexpression of the chemokine receptor CXCR3 and its ligands. Previously, we have demonstrated that pentoxifylline （PTX） alleviated cigarette smoke-induced emphysema. The aim of this study was to determine if the overexpression of CXCR3 and its ligand interferon-inducible protein-10 （IP-10） that was elicited by smoke exposure were attenuated by PTX. Methods （1） The study in vitro： a given number of RAW264.7 macrophages with decreasing concentrations of PTX in the culture medium were challenged with cigarette smoke extract （CSE）; （2） The study in vivo： male BALB/c mice were randomized into four groups, i.e., sham-smoke, smoke only, smoke with 2 mg/kg PTX, and smoke with 10 mg/kg PTX. The smoke exposure time was 90 minutes once a day, 6 days a week for 16 weeks. PTX was given intraperitoneally before each episode of smoke exposure. Interferon （IFN）-y and IP-10 in broncho-alveolar lavage fluid （BALF） and in culture medium were measured by enzyme-linked immunosorbent assay （ELISA）. IP-10 mRNA in lung tissue was assessed by RT-PCR. CXCR3 positive cells in lung sections were visualized by immunochemistry staining. Results Up-regulation of IFN-γ and IP-10 in the culture medium of macrophages elicited by CSE was inhibited by PTX in a dose-dependent manner. Chronic cigarette smoke exposure led to overexpression of IFN-γ and IP-10 in BALF, upregulation of IP-10 mRNA and increased infiltration of CXCR3^＋ cells into lung parenchyma. Administration of PTX decreased the level of IFN-y from （6.26±1.38） ng/ml to （4.43±0.66） ng/ml by low dose PTX or to （1.74±0.28） ng/ml by high dose PTX. IP-10 was reduced from （10.35±1.49） ng/ml to （8.19±0.79） ng/ml by low dose PTX or to （7.51±0.60） ng/ml by high dose PTX. The expression of IP-10 mRNA was also down-regulated （P 〈0.05）. But only with a high dose of PTX was the ratio of CXCR3^＋ cells decreased; 15.2±7.3 vs. 10.4±1.8 （P 〈0.05）. Conclusion PTX attenuates cigarette smoke-induced overexpression of chemokine receptor CXCR3 and its ligand IP-10, which is relevant to its inhibitory effect on pulmonary emphysema.
