目的 結(jié)論探討采取小劑量脂多糖( LPS) 間斷腹腔注射的方式制作內(nèi)毒素誘導(dǎo)的小鼠ALI和肺纖維化動(dòng)物模型, 了解該方法引起的肺急性炎性反應(yīng)以及纖維化的發(fā)展過程。方法健康C57BL/ 6 小鼠40 只隨機(jī)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(gè)LPS 處理組連續(xù)3 d 腹腔注射LPS( 5 mg /kg) , 分別于各觀察終點(diǎn)( 3 d、2 周和4 周) 取材; 對Con 組連續(xù)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(píng)分了解LPS 刺激后不同時(shí)期小鼠肺組織炎癥和纖維化發(fā)展的過程, 同時(shí)采用RT-PCR 檢測了解肺組織Ⅰ型前膠原和α平滑肌肌動(dòng)蛋白( α-SMA) mRNA 表達(dá)強(qiáng)度, 通過肺組織羥脯氨酸含量測定了解肺間質(zhì)膠原沉積和纖維化程度, 并觀察各組實(shí)驗(yàn)動(dòng)物的存活情況。結(jié)果 小鼠在接受LPS 腹腔注射后3 d 肺組織即出現(xiàn)急性炎性反應(yīng); 注射后2 周, 肺間質(zhì)出現(xiàn)膠原沉積, Ashcroft纖維化評(píng)分增高; 4 周形成明顯肺間質(zhì)纖維化, Ashcroft 纖維化評(píng)分進(jìn)一步升高。肺組織膠原及α-SMA的表達(dá)量自LPS 刺激后3 d 起增高, 4 周后達(dá)到高峰。到觀察終點(diǎn)所有動(dòng)物全部存活。結(jié)果 LPS 誘發(fā)的ALI 和炎性反應(yīng)在感染早期即啟動(dòng)了纖維化進(jìn)程。使用劑量為5 mg/kg的LPS 對于小鼠進(jìn)行連續(xù)3 d 腹腔注射可以成功制作內(nèi)毒素誘導(dǎo)的小鼠ALI 和肺纖維化的動(dòng)物模型, 制作成功率高,動(dòng)物死亡率低, 適合對小動(dòng)物進(jìn)行較長時(shí)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(nèi)毒素誘導(dǎo)的小鼠急性肺損傷肺纖維化動(dòng)物模型. 中國呼吸與危重監(jiān)護(hù)雜志, 2010, 9(1): 76-80. doi: 復(fù)制
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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(dǎo)大鼠急性肺損傷作用的研究.中國呼吸與危重監(jiān)護(hù)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(yī)大學(xué)學(xué)報(bào),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.