間斷腹腔注射脂多糖制作內毒素誘導的小鼠急性肺損傷肺纖維化動物模型_《中國呼吸與危重監護雜志》

作者：

何征宇 , 朱也森 , 姜虹

上海交通大學醫學院附屬第九人民醫院麻醉科（上海 200011）通訊作者: 姜虹, E-mail: jianghonjy@ yahoo. com. cn;

關鍵詞：

急性肺損傷肺纖維化脂多糖小鼠動物模型

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目的結論探討采取小劑量脂多糖( LPS) 間斷腹腔注射的方式制作內毒素誘導的小鼠ALI和肺纖維化動物模型, 了解該方法引起的肺急性炎性反應以及纖維化的發展過程。方法健康C57BL/ 6 小鼠40 只隨機分為陰性對照組( Con 組) 、LPS-3 d 組、LPS-2 周組和LPS-4 周組, 每組10 只。對三個LPS 處理組連續3 d 腹腔注射LPS( 5 mg /kg) , 分別于各觀察終點( 3 d、2 周和4 周) 取材; 對Con 組連續3 d 腹腔注射等體積生理鹽水, 于注射后3 d 取材。通過HE 染色和Van-Gieson 膠原染色及Ashcroft 肺纖維化評分了解LPS 刺激后不同時期小鼠肺組織炎癥和纖維化發展的過程, 同時采用RT-PCR 檢測了解肺組織Ⅰ型前膠原和α平滑肌肌動蛋白( α-SMA) mRNA 表達強度, 通過肺組織羥脯氨酸含量測定了解肺間質膠原沉積和纖維化程度, 并觀察各組實驗動物的存活情況。結果小鼠在接受LPS 腹腔注射后3 d 肺組織即出現急性炎性反應; 注射后2 周, 肺間質出現膠原沉積, Ashcroft纖維化評分增高; 4 周形成明顯肺間質纖維化, Ashcroft 纖維化評分進一步升高。肺組織膠原及α-SMA的表達量自LPS 刺激后3 d 起增高, 4 周后達到高峰。到觀察終點所有動物全部存活。結果 LPS 誘發的ALI 和炎性反應在感染早期即啟動了纖維化進程。使用劑量為5 mg/kg的LPS 對于小鼠進行連續3 d 腹腔注射可以成功制作內毒素誘導的小鼠ALI 和肺纖維化的動物模型, 制作成功率高,動物死亡率低, 適合對小動物進行較長時間的觀察和研究。

引用本文： 何征宇,朱也森,姜虹. 間斷腹腔注射脂多糖制作內毒素誘導的小鼠急性肺損傷肺纖維化動物模型. 中國呼吸與危重監護雜志, 2010, 9(1): 76-80. doi: 復制

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11.	Marshall RP,Bellingan G,Webb S,et al.Fibroproliferation occurs early in the acute respiratory distress syndrome and impacts on outcome.Am J Respir Crit Care Med,2000,162:1783-1788.
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1. 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.
2. Feng Y,Yang Q,Xu J,et al.Effects of HMGB1 on PMN apoptosis during LPS-induced acute lung injury.Exp Mol Pathol,2008,85:214-222.
3. 劉虹,安友仲,李芳芳,等.烏司他丁對脂多糖誘導大鼠急性肺損傷作用的研究.中國呼吸與危重監護雜志,2008,7:290-293.
4. 李琦,錢桂生,張青,等.遞增劑量脂多糖致傷對大鼠SIRS-肺損傷的影響.第三軍醫大學學報,2001,23:1264-1267.
5. Zhang XY,Shimura S,Masuda T,et al.Antisense oligonucleotides to NF-kappaB improve survival in bleomycin-induced pneumopathy of the mouse.Am J Respir Crit Care Med,2000,162:1561-1568.
6. Meijerink J,Mandigers C,van de Locht L,et al.A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR.J Mol Diagn,2001,3:55-61.
7. Olman MA,White KE,Ware LB,,et al.Pulmonary edema fluid from patients with early lung injury stimulates fibroblast proliferation through IL-1 beta-induced IL-6 expression.J Immunol ,2004,172:2668-2677.
8. Seki E,De Minicis S,Osterreicher CH,et al.TLR4 enhances TGF-beta signaling and hepatic fibrosis.Nat Med,2007,13:1324-1332.
9. Vancheri C,Crimi N,Conte E,,et al.Human lung fibroblasts inhibit tumor necrosis factor-alpha production by LPS-activated monocytes.Am J Respir Cell Mol Biol,1996,15:460-466.
10. He Z,Zhu Y,Jiang H.Toll-like receptor 4 mediates lipopolysaccharide-induced collagen secretion by phosphoinositide3-kinase-Akt pathway in fibroblasts during acute lung injury.J Recept Signal Transduct Res,2009,29:119-125.
11. Marshall RP,Bellingan G,Webb S,et al.Fibroproliferation occurs early in the acute respiratory distress syndrome and impacts on outcome.Am J Respir Crit Care Med,2000,162:1783-1788.
12. Armstrong L,Thickett DR,Mansell JP,et al.Changes in collagen turnover in early acute respiratory distress syndrome.Am J Respir Crit Care Med,1999,160:1910-1915.

《中國呼吸與危重監護雜志》

間斷腹腔注射脂多糖制作內毒素誘導的小鼠急性肺損傷肺纖維化動物模型

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《中國呼吸與危重監護雜志》

間斷腹腔注射脂多糖制作內毒素誘導的小鼠急性肺損傷肺纖維化動物模型

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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