目的 觀察香煙煙霧提取物( CSE) 對轉(zhuǎn)化生長因子β1 ( TGF-β1 ) 刺激的人肺成纖維細(xì)胞( HLF) 增殖及分泌過氧化氫( H2O2 ) 的影響, 探討CSE 可能參與肺纖維化發(fā)病的機(jī)制。方法 將體外分離培養(yǎng)的HLF 細(xì)胞分為對照組和TGF-β1 ( 5 ng/mL) 刺激組, 用不同濃度的CSE( 0% 、2. 5% 、5% 、10% ) 進(jìn)行干預(yù)。應(yīng)用Brdu ELISA 法檢測細(xì)胞增殖; 熒光分光光度法測定細(xì)胞分泌的H2O2 ; 免疫熒光標(biāo)記分泌的H2O2 和細(xì)胞α-平滑肌肌動蛋白( α-SMA) 表達(dá)。結(jié)果 TGF-β1 刺激組細(xì)胞經(jīng)免疫熒光標(biāo)記顯示α-SMA 陽性表達(dá), 說明HLF 在TGF-β1 刺激下轉(zhuǎn)化為肌成纖維細(xì)胞( MF) 。TGF-β1刺激組中, 與0% 濃度CSE 比較, 2. 5%、5% 濃度的CSE 干預(yù)可以顯著促進(jìn)細(xì)胞增殖( P lt; 0. 01 或0. 05) , 10% 濃度的CSE 干預(yù)抑制細(xì)胞增殖( P lt;0. 01) 。對照組中, 與0% 濃度CSE 比較, 2. 5%、5% 、10% 濃度的CSE 均使細(xì)胞增殖顯著減弱( P lt;0. 01 或P lt;0. 05) , 并呈劑量依賴性。TGF-β1 刺激HLF細(xì)胞能產(chǎn)生一定量的H2O2, CSE 干預(yù)后分泌產(chǎn)生的H2O2 明顯增加( P lt; 0. 01) , 經(jīng)NADPH 氧化酶抑制劑二苯基碘( DPI) 預(yù)處理后檢測不到H2O2。免疫熒光標(biāo)記顯示分泌的H2O2 來源于α-SMA 陽性表達(dá)的MF 細(xì)胞。結(jié)論 低濃度的CSE 能夠促進(jìn)TGF-β1 刺激HLF 細(xì)胞轉(zhuǎn)化的MF細(xì)胞增殖, 并顯著增加MF 細(xì)胞向細(xì)胞外分泌H2O2 , 提示CSE 可能通過氧化應(yīng)激參與肺纖維化的發(fā)生、發(fā)展。
引用本文： 劉寅,張德平. 香煙煙霧提取物對轉(zhuǎn)化生長因子β1 刺激的人肺成纖維細(xì)胞增殖及分泌過氧化氫的影響. 中國呼吸與危重監(jiān)護(hù)雜志, 2009, 09(4): 360-364. doi: 復(fù)制
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| 1. | 周玉民, 王辰, 姚婉貞, 等. 職業(yè)接觸粉塵和煙霧對慢性阻塞性肺疾病及呼吸道癥狀的影響. 中國呼吸與危重監(jiān)護(hù)雜志, 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. | 張德平, 莊誼, 殷凱生. 肌成纖維細(xì)胞凋亡與肺纖維化. 國際呼吸雜志, 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, 姚婉貞, 等. 職業(yè)接觸粉塵和煙霧對慢性阻塞性肺疾病及呼吸道癥狀的影響. 中國呼吸與危重監(jiān)護(hù)雜志, 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. 張德平, 莊誼, 殷凱生. 肌成纖維細(xì)胞凋亡與肺纖維化. 國際呼吸雜志, 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.