Effects of Cigarette Smoke Extract on the Proliferation and Secretion of Hydrogen Peroxide in Human Lung Fibroblasts Induced by Transforming Growth Factor-β1_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

LIU Yin ,  ZHANG Deping.

Department of Respiratory Medicine, The Affiliated Drum Tower Hospital, Nanjing University Medical College.Nanjing, Jiangsu, 210008, China;

Corresponding author：

ZHANG Deping., Email: dpzhang207@yahoo.com.cn

Keywords：

Cigarette smoke extract; Pulmonary fribrosis; Lung fibroblast; Transforming growth factor-β1; Hydrogen peroxide; Oxidative stress; α-Smooth muscle actin

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To observe the effects of cigarette smoke extract ( CSE) on the proliferation and secretion of hydrogen peroxide ( H2O2 ) in human lung fibroblasts ( HLFs) induced by transforming growth factor-β1 ( TGF-β1 ) . Methods Cultured HLFs were divided into a normal group and a model group induced by TGF-β1 ( 5 ng/mL) , then intervened with CSE at different concentrations ( 0% , 2. 5% , 5% ,10% , respectively) . Brdu ELISA assay was used to detect cell proliferation. H2O2 release from cultured cells was assayed using a fluorimetric method. Cellular localization of H2O2 and expression of α-SMA were performed using a fluorescent-labeling strategy. Results TGF-β1 stimulated group showed positive expression of α-SMA, implying TGF-β1 had induced fibroblasts to differentiate into myofibroblasts. In TGF-β1 stimulated group, 2. 5% and 5% CSE promoted cell proliferation ( P lt; 0. 01 or 0. 05) , while 10% CSE inhibited cell proliferation ( P lt; 0. 01) . In the normal group, both low and high concentration of CSE inhibited cell proliferation ( P lt; 0. 01 or P lt; 0. 05) , and the inhibition effect was dose-dependent. HLF induced by TGF-β1 generated low constitutive levels of extracellular H2O2 that was markedly enhanced by CSE treatment ( P lt; 0. 01) . Pretreatment with DPI, an inhibitor of NADPH oxidase, abolished secretion of H2O2 . Cellular localization of H2O2 by a fluorescent-labeling strategy demonstrated that extracellular secretion of H2O2 is specific to the myofibroblast. Conclusions Low concentration of CSE can promote myofibroblast proliferation, and markedly increase extracellular secretion of H2O2 . CSE possibly take part in the development and progress of idiopathic pulmonary fibrosis by increasing oxidative stress.

Citation： LIU Yin,ZHANG Deping.. Effects of Cigarette Smoke Extract on the Proliferation and Secretion of Hydrogen Peroxide in Human Lung Fibroblasts Induced by Transforming Growth Factor-β1. Chinese Journal of Respiratory and Critical Care Medicine, 2009, 09(4): 360-364. doi: Copy

1.	周玉民, 王辰, 姚婉贞, 等. 职业接触粉尘和烟雾对慢性阻塞性肺疾病及呼吸道症状的影响. 中国呼吸与危重监护杂志, 2009,8: 6-11.
2.	Ryu JH, Colby TV, Hartman TE, et al. Smoking-related interstitial lung diseases: a concise review. Eur Respir J, 2001, 17: 122-132.
3.	Carnevali S, Petruzzelli S, Longoni B, et al. Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts. Am J Physiol Lung Cell Mol Physiol, 2003, 284: L955-L963.
4.	Thannickal VJ, Fanburg BL. Activation of an H2O2 -generating NADH oxidase in human lung fibroblasts by transforming growth factor beta 1. J Biol Chem, 1995, 270: 30334-30338.
5.	Selman M, Pardo A. Idiopathic pulmonary fibrosis: an epithelia /fibroblastic cross-talk disorder. Respir Res, 2002, 3 : 3.
6.	Nagai S, Hoshino Y, Hayashi M, et al. Smoking-related interstitial lung diseases. Curr Opin Pulm Med, 2000, 6: 415-419.
7.	Kinnula VJ, Fattman CL, Tan RJ, et al. Oxidative stress in pulmonary fibrosis: a possible role for redox modulatory therapy. Am J Respir Crit Med, 2005, 172: 417-422.
8.	Kinnula VL. Redox imbalance and lung fibrosis. Antioxid Redox Signal, 2008, 10: 249-252.
9.	Waghray M, Cui Z, Horowitz JC, et al. Hydrogen peroxide is a diffusible paracrine signal for the induction of epithelial cell death by activated myofibroblasts. FASEB J, 2005, 19: 854-856.
10.	Phan SH. The myofibroblast in pulmonary fibrosis. Chest, 2002 ,122: 286S-289S.
11.	张德平, 庄谊, 殷凯生. 肌成纤维细胞凋亡与肺纤维化. 国际呼吸杂志, 2007, 27: 298-301.
12.	Ishii T, Matsuse T, Igarashi H, et al. Tobacco smoke reduces viability in human lung fibroblasts: protective effect of glutathione S-transferase P1 . Am J Physiol Lung Cell Mol Physiol, 2001 , 280 :L1189-L1195.
13.	Thannickal VJ, Fanburg BL. Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol, 2000, 279: L1005-L1028.

1. 周玉民, 王辰, 姚婉贞, 等. 职业接触粉尘和烟雾对慢性阻塞性肺疾病及呼吸道症状的影响. 中国呼吸与危重监护杂志, 2009,8: 6-11.
2. Ryu JH, Colby TV, Hartman TE, et al. Smoking-related interstitial lung diseases: a concise review. Eur Respir J, 2001, 17: 122-132.
3. Carnevali S, Petruzzelli S, Longoni B, et al. Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts. Am J Physiol Lung Cell Mol Physiol, 2003, 284: L955-L963.
4. Thannickal VJ, Fanburg BL. Activation of an H2O2 -generating NADH oxidase in human lung fibroblasts by transforming growth factor beta 1. J Biol Chem, 1995, 270: 30334-30338.
5. Selman M, Pardo A. Idiopathic pulmonary fibrosis: an epithelia /fibroblastic cross-talk disorder. Respir Res, 2002, 3 : 3.
6. Nagai S, Hoshino Y, Hayashi M, et al. Smoking-related interstitial lung diseases. Curr Opin Pulm Med, 2000, 6: 415-419.
7. Kinnula VJ, Fattman CL, Tan RJ, et al. Oxidative stress in pulmonary fibrosis: a possible role for redox modulatory therapy. Am J Respir Crit Med, 2005, 172: 417-422.
8. Kinnula VL. Redox imbalance and lung fibrosis. Antioxid Redox Signal, 2008, 10: 249-252.
9. Waghray M, Cui Z, Horowitz JC, et al. Hydrogen peroxide is a diffusible paracrine signal for the induction of epithelial cell death by activated myofibroblasts. FASEB J, 2005, 19: 854-856.
10. Phan SH. The myofibroblast in pulmonary fibrosis. Chest, 2002 ,122: 286S-289S.
11. 张德平, 庄谊, 殷凯生. 肌成纤维细胞凋亡与肺纤维化. 国际呼吸杂志, 2007, 27: 298-301.
12. Ishii T, Matsuse T, Igarashi H, et al. Tobacco smoke reduces viability in human lung fibroblasts: protective effect of glutathione S-transferase P1 . Am J Physiol Lung Cell Mol Physiol, 2001 , 280 :L1189-L1195.
13. Thannickal VJ, Fanburg BL. Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol, 2000, 279: L1005-L1028.

Next Article
From Trunk to Leaves: Exploration and Practice of Conformity in Eight Years’Medical Education Courses

Chinese Journal of Respiratory and Critical Care Medicine

Effects of Cigarette Smoke Extract on the Proliferation and Secretion of Hydrogen Peroxide in Human Lung Fibroblasts Induced by Transforming Growth Factor-β1

Abstract Full text Figures/Tables Video References Cited by

Next Article

Format

Content