• 1 復旦大學附屬中山醫(yī)院老年科( 上海 200032);;
  • 2 復旦大學附屬中山醫(yī)院呼吸科( 上海 200032) ;;
  • 3 復旦大學附屬中山醫(yī)院中心實驗室( 上海200032) ;;
  • 4 復旦大學附屬中山醫(yī)院病理科( 上海200032)通訊作者: 朱蕾, E-mail: tfzhu@ 126. com;

目的  研究髓樣分化蛋白2( MD-2) 在脂多糖( LPS) 誘導的急性肺損傷( ALI) 大鼠肺組織中的表達及其作用。方法  20 只健康雄性SD 大鼠隨機分為LPS 組和對照組, 通過支氣管肺泡灌洗分離肺泡巨噬細胞, 采用RT-PCR、Western blot 和免疫組化方法分別檢測MD-2 mRNA 和蛋白的表達, ELISA 方法測定血清腫瘤壞死因子α( TNF-α) 水平。將MD-2 siRNA 轉(zhuǎn)染大鼠肺泡巨噬細胞系NR8383, 以終濃度1 μg/mL的LPS 刺激細胞, 采用RT-PCR 和Western blot 方法分別測定細胞MD-2mRNA 和蛋白的表達, ELISA 方法測定細胞培養(yǎng)上清液TNF-α水平。結(jié)果  LPS 組大鼠肺泡巨噬細胞和肺組織標本中MD-2 mRNA 和蛋白的表達均顯著高于對照組( P  lt;0. 01) , 血清TNF-α水平明顯升高( P  lt;0. 01) 。在LPS 刺激下, NR8383 細胞MD-2 mRNA 和蛋白的表達顯著增加( P  lt;0. 01) , 細胞培養(yǎng)上清液中TNF-α水平升高( P  lt;0. 01) 。經(jīng)MD-2 siRNA 處理后, NR8383 細胞在LPS 刺激下MD-2
mRNA 和蛋白的表達未見明顯增加( P  gt; 0. 05) , 細胞培養(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 大鼠肺組織中表達顯著上調(diào), 沉默肺泡巨噬細胞MD-2 基因可抑制LPS 刺激引起的細胞因子分泌增加, 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. 中國呼吸與危重監(jiān)護雜志, 2010, 9(6): 614-618. doi: 復制

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. 在實踐中提高對急性肺損傷和急性呼吸窘迫綜合征新概念的認識. 中國呼吸與危重監(jiān)護雜志, 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. 不同劑量脂多糖對大鼠急性肺損傷效應(yīng)的觀察. 第三軍醫(yī)大學學報, 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 基因表達的影響. 中國危重病急救醫(yī)學雜志, 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 型上皮細胞中的表達及作用. 重慶醫(yī)學, 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, 張進川, 趙亞力, 等. 地塞米松對急性肺損傷大鼠炎癥介質(zhì)/抗炎介質(zhì)表達的影響. 中國呼吸與危重監(jiān)護雜志, 2005 ,4: 65-68.
  1. 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. 2. 錢桂生. 在實踐中提高對急性肺損傷和急性呼吸窘迫綜合征新概念的認識. 中國呼吸與危重監(jiān)護雜志, 2002 , 1: 199-201 .
  3. 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. 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. 5. 李琦, 錢桂生, 張青, 等. 不同劑量脂多糖對大鼠急性肺損傷效應(yīng)的觀察. 第三軍醫(yī)大學學報, 2004, 26: 871-873 .
  6. 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. 7. 杜曉輝, 張永明, 李榮, 等. 性別差異對膿毒癥大鼠肺組織Toll樣受體4 及髓樣分化蛋白-2 基因表達的影響. 中國危重病急救醫(yī)學雜志, 2005, 17: 726-729.
  8. 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. 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. 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. 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. 12. 李永旺, 張德明, 錢桂生, 等.MD2 在內(nèi)毒素激活肺泡II 型上皮細胞中的表達及作用. 重慶醫(yī)學, 2007, 36: 1582-1584.
  13. 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. 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. 15. 李洪霞, 張進川, 趙亞力, 等. 地塞米松對急性肺損傷大鼠炎癥介質(zhì)/抗炎介質(zhì)表達的影響. 中國呼吸與危重監(jiān)護雜志, 2005 ,4: 65-68.