Mechanical Stretch Activates Alveolar Macrophages to Produce Cytokines and Has Synergistic Effect with Lipopolysaccharide in Inducing MIP-2 Release_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

DING Ning ¹ , XIAO Hui ² ,  XU Lixin ¹ , SHE Shouzhang. ¹

Department of Anesthesiology, The First Municipal People’s Hospital of Guangzhou. Guangzhou,Guangdong, 510180, China;

Corresponding author：

XU Lixin, Email: gzxulixin@163.com

Keywords：

Mechanical stretch; Alveolar macrophage; Cytokine; Macrophage inflammatory protein-2; Ventilator-induced lung injury

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Objective To observe the effects of mechanical stretch on cytokines release from alveolar macrophages( AMs) and the expression of macrophage inflammatory protein-2( MIP-2) induced by lipopolysaccharide( LPS) . Methods AMs were divided into the following groups: ①AMs were subjected to 20% elongation by Flexercell 4000T cell stress system for 24 hours and the supernatant was collected to detect the levels of TNF-α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IFN-γ, macrophage inflammatory protein-1α( MIP-1α) , MIP-2, monocyte chemoattractant protein-1( MCP-1) , granulocyte /macrophage colony stimulating factors( GM-CSF) , interferon inducible protein-10( IP-10) , regulated on activation in normal T-cell expressed and secreted( Rantes) and keratinocyte chemoattractant( KC) , by using LiquiChip system. ② AMs were subjected to 5% , 10% , 15% and 20% elongation for 24 hours and the supernatant was collected to detect the levels of MIP-2. ③AMs were subjected to 20% elongation and MIP-2 in supernatant was detected 1, 3,6, 12, and 24 hours later. ④ AMs were subjected to 20% elongation and/ or LPS at a concentration of 10 ng/mL, and MIP-2 in supernatant was detected 24 hours later. Unstretched AMs were used as control in all kind of test. Results ①The levels of IL-1β, IL-6,MIP-2, MCP-1, IFN-γand IP-10 secreted by stretched AMs were 8. 7, 4. 3, 38. 6, 4. 8, 14. 2 and 5. 0 times those of the control group( all P lt; 0. 001) . ② The levels of MIP-2 secreted by AMs subjected to 10% , 15% and 20% elongation were ( 480. 5 ±93. 1) pg /mL,( 806. 3 ±225. 9) pg/mL and ( 1335. 7 ±18. 5) pg/mL respectively, all significantly higher than those oft he control group [ ( 34. 6 ±11. 4) pg/mL, all P lt;0. 001] . ③ Three hours after the stimulation of stretch the level of MIP-2 began to increase gradually. And 6, 12, and 24 hours after the stimulation the levels of MIP-2 secreted by the AMs were ( 819. 4 ±147. 5) pg/mL, ( 1287. 6 ±380 ±3 ) pg/mL and ( 1455. 9 ±436. 7) pg/mLrespectively, all significantly higher than those of the control group[ ( 33. 4 ±10. 2) pg/mL, all P lt; 0. 001] . ④When the AMs were stimulated individually by LPS( 10 ng /mL) or mechanical stretch ( 20% ) , the levels of MIP-2 increased to ( 1026. 3 ±339. 5 ) pg/mL and ( 1335. 7 ±318. 5 ) pg/mL respectively( both P lt; 0. 001) . When the AMs were costimulated by LPS and mechanical stretch, the level of MIP-2 increased to ( 2275. 3 ±492. 1) pg/mL, implicating a synergistic effect between mechanical stretch and LPS ( F = 121. 983, P lt; 0. 001) . Conclusions Mechanical stretch activates AMs to produce multiple inflammatory cytokines and induce AMs to secret MIP-2 in a strength- and time-dependent manner.Mechanical stretch also has synergistic effect with LPS in inducing MIP-2 release, which might play an important role in the development of ventilator-induced lung injury.

Citation： DING Ning ,XIAO Hui,XU Lixin,SHE Shouzhang.. Mechanical Stretch Activates Alveolar Macrophages to Produce Cytokines and Has Synergistic Effect with Lipopolysaccharide in Inducing MIP-2 Release. Chinese Journal of Respiratory and Critical Care Medicine, 2009, 09(2): 166-170. doi: Copy

1.	董春山, 徐志新, 徐瑞好, 等. 机械通气诱导大鼠肺损伤病理及炎性细胞因子的变化. 中国呼吸与危重监护杂志, 2008, 7: 135-139.
2.	秦雪冰, 俞森洋. 呼吸机所致肺损伤家兔核因子-κB 表达的意义. 中国呼吸与危重监护杂志, 2006, 5: 22-24.
3.	丁宁, 肖慧, 高巨, 等. P38 及上游激酶MKK6 对机械牵张应变诱导肺泡巨噬细胞HMGB1 的调控. 生理学报, 2009, 61: 49-55.
4.	Belperio JA, Keane MP, Lynch JP 3rd, et al. The role of cytokines during the pathogenesis of ventilator-associated and ventilatorinduced lung injury. Semin Respir Crit Care Med,2006, 27: 350-364.
5.	Frank J,Wray C, McAuley D, et al. Alveolar macrophages contribute to alveolar epithelial barrier dysfunction in ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol, 2006 , 291: L1191 -L1198.
6.	Oeckler RA, Hubmayr RD. Ventilator-associated lung injury: a search for better therapeutic targets. Eur Respir J, 2007, 30: 1216-1226.
7.	丁宁, 许立新, 郑彬, 等. 体外培养大鼠肺泡巨噬细胞周期性牵张模型的建立及其炎性介质的变化. 中华生物医学工程杂志,2008, 14: 18-21.
8.	Johannisson A, Jonasson R, Dernfalk J, et al. Simultaneous detection of porcine proinflammatory cytokines using multiplex flow cytometry by the xMAP technology. Cytometry A, 2006 , 69: 391 -395.
9.	Pugin J, Dunn I, Jolliet P, et al. Activation of human macrophages by mechanical ventilation in vitro. Am J Physiol, 1998, 275: L1040-L1050.
10.	Eyal FG, Hamm CR, Parker JC. Reduction in alveolar macrophages attenuates acute ventilator induced lung injury in rats. Intensive Care Med, 2007, 33: 1212-1218.
11.	Ning QM,Wang XR. Response of alveolar type II epithelial cells to mechanical stretch and lipopolysaccharide. Respiration, 2007, 74 :579-585.

1. 董春山, 徐志新, 徐瑞好, 等. 机械通气诱导大鼠肺损伤病理及炎性细胞因子的变化. 中国呼吸与危重监护杂志, 2008, 7: 135-139.
2. 秦雪冰, 俞森洋. 呼吸机所致肺损伤家兔核因子-κB 表达的意义. 中国呼吸与危重监护杂志, 2006, 5: 22-24.
3. 丁宁, 肖慧, 高巨, 等. P38 及上游激酶MKK6 对机械牵张应变诱导肺泡巨噬细胞HMGB1 的调控. 生理学报, 2009, 61: 49-55.
4. Belperio JA, Keane MP, Lynch JP 3rd, et al. The role of cytokines during the pathogenesis of ventilator-associated and ventilatorinduced lung injury. Semin Respir Crit Care Med,2006, 27: 350-364.
5. Frank J,Wray C, McAuley D, et al. Alveolar macrophages contribute to alveolar epithelial barrier dysfunction in ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol, 2006 , 291: L1191 -L1198.
6. Oeckler RA, Hubmayr RD. Ventilator-associated lung injury: a search for better therapeutic targets. Eur Respir J, 2007, 30: 1216-1226.
7. 丁宁, 许立新, 郑彬, 等. 体外培养大鼠肺泡巨噬细胞周期性牵张模型的建立及其炎性介质的变化. 中华生物医学工程杂志,2008, 14: 18-21.
8. Johannisson A, Jonasson R, Dernfalk J, et al. Simultaneous detection of porcine proinflammatory cytokines using multiplex flow cytometry by the xMAP technology. Cytometry A, 2006 , 69: 391 -395.
9. Pugin J, Dunn I, Jolliet P, et al. Activation of human macrophages by mechanical ventilation in vitro. Am J Physiol, 1998, 275: L1040-L1050.
10. Eyal FG, Hamm CR, Parker JC. Reduction in alveolar macrophages attenuates acute ventilator induced lung injury in rats. Intensive Care Med, 2007, 33: 1212-1218.
11. Ning QM,Wang XR. Response of alveolar type II epithelial cells to mechanical stretch and lipopolysaccharide. Respiration, 2007, 74 :579-585.

Chinese Journal of Respiratory and Critical Care Medicine

Mechanical Stretch Activates Alveolar Macrophages to Produce Cytokines and Has Synergistic Effect with Lipopolysaccharide in Inducing MIP-2 Release

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