小鼠急性肺损伤造模条件的探究

doi:10.16431/j.cnki.1671-7236.2017.08.008

摘要/Abstract

摘要：

为探究构建急性肺损伤模型的条件，本试验选用健康昆明小鼠，经腹腔注射递增剂量脂多糖（LPS）（0、2、4、6、8、10 mg/kg），观察不同条件下小鼠呼吸频率、死亡率、肺脏干湿重比值及组织结构变化，检测炎症因子IL-10、TNF-α、IL-1β及IFN-γ在不同程度肺损伤中的表达量变化。结果显示，4 mg/kg LPS组肺脏干湿重比值显著高于对照组（P<0.05）；6、8和10 m/kg LPS组肺脏干湿重比值极显著高于对照组（P<0.01）。病理切片分析显示，6 mg/kg LPS组小鼠肺脏出现充血，肺泡腔及肺间质出现渗出，肺泡隔增厚，出现少量中性粒细胞浸润；8、10 mg/kg LPS组小鼠肺脏充血明显，肺泡腔及肺间质出现渗出，肺泡明显隔增厚，出现中性粒细胞浸润；与对照组相比，6、8、10 mg/kg LPS组肺组织中胶原纤维大量增多，且多出现在肺气管周围，10 mg/kg LPS组现象最明显。炎症因子检测分析结果显示，与对照组相比，2、4、6、8、10 mg/kg LPS组炎症因子均极显著增加（P<0.01）。结合病理组织切片及相关检测指标表明，昆明小鼠腹腔注射8 mg/kg LPS并作用12 h，可成功构建急性肺损伤模型。

关键词: 急性肺损伤; 小鼠; 脂多糖; 肺组织

Abstract:

In order to explore the building conditions of acute lung injury, healthy Kunming mice were selected, by intraperitoneal injection of cascade dose of LPS (0, 2, 4, 6, 8, 10 mg/kg) to observe the mice respiratory frequency, mortality, lung W/D value and the changes of tissue structure in different conditions, and detect the expression changes of inflammation cytokine, IL-10,TNF-α, IL-1β and IFN-γ. The results showed that compared with control group, lung W/D value in 4 mg/kg LPS group significantly increased (P<0.05) and lung W/D values in 6, 8 and 10 mg/kg LPS groups extremely significantly increased (P<0.01). Pathological analysis showed that 6 mg/kg LPS group appeared lung congestion, alveolar luminal and pulmonary interstitial exudation, alveolar septal thickening, and a small amount of neutrophil infiltration. 8 and 10 mg/kg LPS groups appeared pulmonary congestion, alveolar and pulmonary interstitial exudation, alveolar septal thickening and neutrophil infiltration. Compared with control group, collagen fibers of the lung tissue in 6, 8, 10 mg/kg LPS group increased, and more appeared in the lung around the trachea, 10 mg/kg LPS group was the most obvious. The inflammatory factors in 2, 4, 6, 8 and 10 mg/kg LPS groups had extremely significantly increased compared with control group (P<0.01). Pathological biopsy and related indicators showed that the model of mice could be successfully constructed by intraperitoneal injection of 8 mg/kg LPS for 12 h.

Key words: acute lung injury; mouse; lipopolysaccharide (LPS); lung tissue

中图分类号:

S852.1

罗友, 吴欣瞳, 庞欣欣, 肖珂, 彭克美, 宋卉. 小鼠急性肺损伤造模条件的探究[J]. 《中国畜牧兽医》, 2017, 44(8): 2269-2276.

LUO You, WU Xin-tong, PANG Xin-xin, XIAO Ke, PENG Ke-mei, SONG Hui. Study on the Building Conditions of Acute Lung Injury in Mice[J]. , 2017, 44(8): 2269-2276.

参考文献

[1] 隰建成, 周宝桐. 脓毒症过程中的炎症介质与细胞凋亡关系的研究进展[J]. 中国危重病急救医学, 2000, 12(6):378-380.
[2] Wang J, Liu Y T, Xiao L, et al. Anti-inflammatory effects of apigenin in lipopolysaccharide-induced inflammatory in acute lung injury by suppressing COX-2 and NF-κB pathway[J]. Inflammation, 2014, 37(6):2085-2090.
[3] Idell S. Coagulation, fibrinolysis, and fibrin deposition in acute lung injury[J].Crit Care Med, 2003, 31(4 Suppl):S213-S220.
[4] Gunther A, Mosavi P, Heinemann S, et al. Alveolar fibrin formation caused by enhanced pneumonia. Comparison and depressed fibrinolytic capacities in severe with the acute respiratory distress syndrome[J]. Am J Respir Crit Care Med, 2000, 161(2 Pt 1):454-462.
[5] Zhong W, Cui Y, Yu Q, et al. Modulation of LPS-stimulated pulmonary inflammation by borneol in murine acute lung injury model[J]. Inflammation, 2014, 37(4):1148-1157.
[6] Bhandary Y P, Shetty S K, Marudamuthu A S, et al. Role of p53-fibrinolytic system cross-talk in the regulation of quartz-induced lung injury[J]. Toxicology and Applied Pharmacology, 2015, 283(2):92-98.
[7] 牛星. 牛肺炎的防治[J]. 中国畜牧兽医文摘, 2016,3:171.
[8] 方青, 高荣, 高英杰, 等. 急性肺损伤动物模型的研究现状[J]. 中国畜牧兽医, 2010,37(5):50-54.
[9] Fei A M,Calalang-Colucci M C. Acute lung injury/acute respiratory distress syndrome in sepsis and septic shock[J].Crit Care, 2000,16(2):289-317.
[10] Downey G P,Dong Q,Kruger J, et al. Regulation of neutrophil activation in acute lung injury[J].Chest,1999,116(1 Suppl):46S-54S.
[11] Standiford T J, Ward P A. Therapeutic targeting of acute lung injury and acute respiratory distress syndrome[J]. Translational Research, 2016, 167(1):183-191.
[12] Bao S, Zou Y, Wang B, et al. Ginsenoside Rg1 improves lipopolysaccharide-induced acute lung injury by inhibiting inflammatory responses and modulating infiltration of M2 macrophages[J]. International Immunopharmacology, 2015, 28(1):429-434.
[13] Walters D M, Cho H Y, Kleeberger S R. Oxidative stress and antioxidants in the pathogenesis of pulmonary fibrosis:A potential role for nrf2[J]. Antioxidants and Redox Signalling, 2008, 10(2):321-332.
[14] 梁华平,姚咏明,王正国.创伤后脓毒症的治疗现状小容乐观——预警研究值得重视[J].第三军医大学学报,2011, 33(21):2223-2226.
[15] 孟庆雪, 刘玉芬, 刘静, 等. 肺表面活性蛋白研究进展[J]. 中国畜牧兽医, 2014, 41(3):178-181.
[16] Wynn T A. Integrating mechanisms of pulmonary fibrosis[J]. Journal of Experimental Medicine, 2011, 208(7):1339-1350.
[17] 邱海波, 陈德昌. 急性肺损伤的炎症反应机制与药物治疗探讨[J]. 中国危重病急救医学, 1999, 11(11):678-680.
[18] Bone R C, Grodzin C J, Balk R A. Sepsis:A new hypothesis for pathogenesis of the disease process[J]. Chest, 1997, 112(1):235-243.
[19] 仇佳明. 积雪草粗提取物对脂多糖诱导小鼠急性肺损伤的治疗作用[D]. 长春:吉林大学, 2015.
[20] Taga K, Tosato G. IL-10 inhibits human T cell proliferation and IL-2 production[J]. The Journal of Immunology, 1992, 148(4):1143-1148.
[21] de Waal Malefyt R, Figdor C G, Huijbens R, et al. Effects of IL-13 on phenotype, cytokine production, and cytotoxic function of human monocytes. Comparison with IL-4 and modulation by IFN-gamma or IL-10[J]. The Journal of Immunology, 1993, 151(11):6370-6381.
[22] Keane M P, Donnelly S C, Belperio J A, et al. Imbalance in the expression of CXC chemokines correlates with bronchoalveolar lavage fluid angiogenic activity and procollagen levels in acute respiratory distress syndrome[J]. The Journal of Immunology, 2002, 169(11):6515-6521.
[23] Wolthuis E K, Vlaar A P J, Choi G, et al. Recombinant human soluble tumor necrosis factor-alpha receptor fusion protein partly attenuates ventilator-induced lung injury[J]. Shock, 2009, 31(3):262-266.
[24] Cross L J M, Matthay M A. Biomarkers in acute lung injury:Insights into the pathogenesis of acute lung injury[J]. Critical Care Clinics, 2011, 27(2):355-377.
[25] Matute-Bello G, Winn R K, Jonas M, et al. Fas(CD95)induces alveolar epithelial cell apoptosis in vivo:Implications for acute pulmonary inflammation[J].Am J Pathol,2001,158:153-161.
[26] 夏道奎,刘云,胡旭.体外循环手术后血浆PAF PAF、IL-8、IFN-γ的变化及其意义[J]. 中国胸心血管外科临床杂志,2014,21(2):212-215.