目的 研究髓樣分化蛋白2( MD-2) 在脂多糖( LPS) 誘導(dǎo)的急性肺損傷( ALI) 大鼠肺組織中的表達(dá)及其作用。方法 20 只健康雄性SD 大鼠隨機(jī)分為L(zhǎng)PS 組和對(duì)照組, 通過支氣管肺泡灌洗分離肺泡巨噬細(xì)胞, 采用RT-PCR、Western blot 和免疫組化方法分別檢測(cè)MD-2 mRNA 和蛋白的表達(dá), ELISA 方法測(cè)定血清腫瘤壞死因子α( TNF-α) 水平。將MD-2 siRNA 轉(zhuǎn)染大鼠肺泡巨噬細(xì)胞系NR8383, 以終濃度1 μg/mL的LPS 刺激細(xì)胞, 采用RT-PCR 和Western blot 方法分別測(cè)定細(xì)胞MD-2mRNA 和蛋白的表達(dá), ELISA 方法測(cè)定細(xì)胞培養(yǎng)上清液TNF-α水平。結(jié)果 LPS 組大鼠肺泡巨噬細(xì)胞和肺組織標(biāo)本中MD-2 mRNA 和蛋白的表達(dá)均顯著高于對(duì)照組( P lt;0. 01) , 血清TNF-α水平明顯升高( P lt;0. 01) 。在LPS 刺激下, NR8383 細(xì)胞MD-2 mRNA 和蛋白的表達(dá)顯著增加( P lt;0. 01) , 細(xì)胞培養(yǎng)上清液中TNF-α水平升高( P lt;0. 01) 。經(jīng)MD-2 siRNA 處理后, NR8383 細(xì)胞在LPS 刺激下MD-2
mRNA 和蛋白的表達(dá)未見明顯增加( P gt; 0. 05) , 細(xì)胞培養(yǎng)上清液中TNF-α水平未見明顯升高( P gt;0. 05) 。結(jié)論 MD-2 在LPS 所致ALI 大鼠肺組織中表達(dá)顯著上調(diào), 沉默肺泡巨噬細(xì)胞MD-2 基因可抑制LPS 刺激引起的細(xì)胞因子分泌增加, 提示MD-2 在LPS所致大鼠ALI 中起重要作用。
引用本文： 任衛(wèi)英 ,華峰,金建軍,曾海英,朱蕾. 髓樣分化蛋白2 在脂多糖所致大鼠急性肺損傷中的作用. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2010, 9(6): 614-618. doi: 復(fù)制
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| 4. | Imai Y, Kaba K, Neely GG, et al. Identification of oxidative stress and Toll-like receptor 4 signaling as a key pathway of acute lung injury. Cell, 2008, 133: 235-249 . |
| 5. | 李琦, 錢桂生, 張青, 等. 不同劑量脂多糖對(duì)大鼠急性肺損傷效應(yīng)的觀察. 第三軍醫(yī)大學(xué)學(xué)報(bào), 2004, 26: 871-873 . |
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| 7. | 杜曉輝, 張永明, 李榮, 等. 性別差異對(duì)膿毒癥大鼠肺組織Toll樣受體4 及髓樣分化蛋白-2 基因表達(dá)的影響. 中國(guó)危重病急救醫(yī)學(xué)雜志, 2005, 17: 726-729. |
| 8. | Everhart MB, Han W, Sherrill TP, et al. Duration and intensity of NF-κB activity determine the severity of endotoxin-induced acute lung injury. J Immunology, 2006, 176: 4995-5005. |
| 9. | Viriyakosol S, Tobias PS, Kirkland TN, et al. Mutational analysis of membrane and soluble forms of human MD-2. J Biol Chem, 2006 ,281: 11955-11964. |
| 10. | Jia HP, Kline JN, Penisten A, et al. Endotoxin responsiveness of human airway epithelia is limited by low expression of MD-2. Am J Physiol, 2004 , 287: L428-437. |
| 11. | Lauer S, Kunde YA, Apodaca TA, et al. Soluble MD2 increases TLR4 levels on the epithelial cell surface. Cell Immunology, 2009 ,255: 8 -16 . |
| 12. | 李永旺, 張德明, 錢桂生, 等.MD2 在內(nèi)毒素激活肺泡II 型上皮細(xì)胞中的表達(dá)及作用. 重慶醫(yī)學(xué), 2007, 36: 1582-1584. |
| 13. | Gropper MA, Wiener-Kronish J. The epithelium in acute lung injury/acute respiratory distress syndrome. Curr Opin Crit Care,2008, 14: 11 -15 . |
| 14. | Togbe D, Schnyder-Candrian S, Schnyder B, et al. Toll-like receptor and tumour necrosis factor dependent endotoxin-induced acute lung injury. Int J Exp Path, 2007, 88: 387-391 . |
| 15. | 李洪霞, 張進(jìn)川, 趙亞力, 等. 地塞米松對(duì)急性肺損傷大鼠炎癥介質(zhì)/抗炎介質(zhì)表達(dá)的影響. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2005 ,4: 65-68. |
- 1. Resman N, Vasl J, Oblak A, et al. Essential roles of hydrophobic residues in both MD-2 and toll-like receptor 4 in activation by endotoxin. J Biol Chem, 2009, 284 : 15052-15060 .
- 2. 錢桂生. 在實(shí)踐中提高對(duì)急性肺損傷和急性呼吸窘迫綜合征新概念的認(rèn)識(shí). 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2002 , 1: 199-201 .
- 3. Baumgarten G, Knuefermann P, Wrigge H, et al. Role of Toll-like receptor 4 for the pathogenesis of acute lung injury in Gram-negative sepsis. Eur J Anaesthesiol, 2006 , 23: 1041-1048.
- 4. Imai Y, Kaba K, Neely GG, et al. Identification of oxidative stress and Toll-like receptor 4 signaling as a key pathway of acute lung injury. Cell, 2008, 133: 235-249 .
- 5. 李琦, 錢桂生, 張青, 等. 不同劑量脂多糖對(duì)大鼠急性肺損傷效應(yīng)的觀察. 第三軍醫(yī)大學(xué)學(xué)報(bào), 2004, 26: 871-873 .
- 6. Oshikawa K, Sugiyama Y. Gene expression of Toll-like receptors and associated molecules induced by inflammatory stimuli in the primary alveolar macrophage. Bio Bio Res Com, 2003, 305 : 649-655.
- 7. 杜曉輝, 張永明, 李榮, 等. 性別差異對(duì)膿毒癥大鼠肺組織Toll樣受體4 及髓樣分化蛋白-2 基因表達(dá)的影響. 中國(guó)危重病急救醫(yī)學(xué)雜志, 2005, 17: 726-729.
- 8. Everhart MB, Han W, Sherrill TP, et al. Duration and intensity of NF-κB activity determine the severity of endotoxin-induced acute lung injury. J Immunology, 2006, 176: 4995-5005.
- 9. Viriyakosol S, Tobias PS, Kirkland TN, et al. Mutational analysis of membrane and soluble forms of human MD-2. J Biol Chem, 2006 ,281: 11955-11964.
- 10. Jia HP, Kline JN, Penisten A, et al. Endotoxin responsiveness of human airway epithelia is limited by low expression of MD-2. Am J Physiol, 2004 , 287: L428-437.
- 11. Lauer S, Kunde YA, Apodaca TA, et al. Soluble MD2 increases TLR4 levels on the epithelial cell surface. Cell Immunology, 2009 ,255: 8 -16 .
- 12. 李永旺, 張德明, 錢桂生, 等.MD2 在內(nèi)毒素激活肺泡II 型上皮細(xì)胞中的表達(dá)及作用. 重慶醫(yī)學(xué), 2007, 36: 1582-1584.
- 13. Gropper MA, Wiener-Kronish J. The epithelium in acute lung injury/acute respiratory distress syndrome. Curr Opin Crit Care,2008, 14: 11 -15 .
- 14. Togbe D, Schnyder-Candrian S, Schnyder B, et al. Toll-like receptor and tumour necrosis factor dependent endotoxin-induced acute lung injury. Int J Exp Path, 2007, 88: 387-391 .
- 15. 李洪霞, 張進(jìn)川, 趙亞力, 等. 地塞米松對(duì)急性肺損傷大鼠炎癥介質(zhì)/抗炎介質(zhì)表達(dá)的影響. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2005 ,4: 65-68.