[1]张利鹏,赵 焱,刘国娟,等.乌司他丁通过干预p38MAPK/ERK信号通路减轻脓毒症性肺损伤[J].中国药理学通报,2016,(09):1311-1316.[doi:10.3969/j.issn.1001-1978.2016.09.024]
 ZHANG Li-peng,ZHAO Yan,LIU Guo-juan,et al.Ulinastatin attenuates lipopolysaccharide-induced acute lung injury by intervening p38MAPK/ERK signaling pathway[J].Chinese Pharmacological Bulletin,2016,(09):1311-1316.[doi:10.3969/j.issn.1001-1978.2016.09.024]
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乌司他丁通过干预p38MAPK/ERK信号通路减轻脓毒症性肺损伤()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2016年09期
页码:
1311-1316
栏目:
论著
出版日期:
2016-09-15

文章信息/Info

Title:
Ulinastatin attenuates lipopolysaccharide-induced acute lung injury by intervening p38MAPK/ERK signaling pathway
文章编号:
1001-1978(2016)09-1311-06
作者:
张利鹏1赵 焱2刘国娟1杨大刚1周丽华1
1.内蒙古医科大学附属医院重症医学科,内蒙古 呼和浩特 010050;
2.内蒙古自治区人民医院神经内科,内蒙古 呼和浩特 010051
Author(s):
ZHANG Li-pengZHAO YanLIU Guo-juanYANG Da-gangZHOU Li-hua
1.ICU,the Affiliated Hospital of Innner Mongolia Medical University,Huhhot 010050,China;
2.Dept of Neurology,the People's Hospital of Innner Mongolia,Huhhot 010051,China
关键词:
乌司他丁 细菌脂多糖 急性呼吸窘迫综合征 p38丝裂原活化蛋白激酶 细胞外信号调节蛋白激酶 减轻
Keywords:
ulinastatin lipopolysaccharide acute respiratory distress syndrome p38MAPK ERK attenuate
分类号:
R-332;R332.35;R329.25;R345.57;R563.8;R631;R977.3
DOI:
10.3969/j.issn.1001-1978.2016.09.024
文献标志码:
A
摘要:
目的 探讨乌司他丁(ulinastatin,UTI)对脓毒症性急性呼吸窘迫综合征(acute respiratory distress syndrome, ARDS)大鼠的肺保护作用及可能机制。方法 56只Wistar大鼠按随机数字表法分为对照组、模型(LPS 6、12、24 h)组、乌司他丁(UTI 6、12、24 h)组,每组8只。采用腹腔注射细菌脂多糖(lipopolysaccharide,LPS,10 mg·kg-1)的方法制备大鼠ARDS模型,乌司他丁组在左侧腹腔注射LPS后1 h后,在右侧腹腔注射乌司他丁20 000 U·kg-1,对照组、注射同等剂量的生理盐水,各组大鼠在不同时间点留取肺组织及血浆标本。计算肺组织湿/干质量(W/D)比值; 光镜下观察肺组织病理改变; 用ELISA法检测各组血浆中肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)、白细胞介素-18(interleukin-18, IL-18)、肺表面活性物质A((surfactant protein A,SPA)含量,检测各组肺匀浆中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide Dismutase,SOD)和一氧化氮(nitric oxide, NO)含量; 免疫组化法检测p38丝裂原活化蛋白激酶(mitogen activated protein kinase, p38MAPK)、细胞外信号调节蛋白激酶(extracellular-signal regulated protein kinase,ERK)蛋白表达情况; Western blot法检测肺组织中磷酸化p38MAPK(p-p38MAPK)及磷酸化ERK(p-ERK)蛋白表达的变化。结果 光镜下观察对照组肺组织结构完整,肺泡腔清晰; LPS组肺泡壁增厚,渗出明显,肺组织出现损伤性变化,以12h组变化最明显; UTI组各组病理改变较LPS组明显减轻。与对照组比较,LPS组W/D比值、血浆TNF-α、IL-18含量,肺匀浆MDA、NO含量均明显升高(P<0.05),而肺匀浆SOD含量明显降低; 与LPS组比较,UTI各组肺组织W/D比值、血浆TNF-α、IL-18含量,肺匀浆MDA含量均明显降低,而肺匀浆SOD含量明显增高,差异有统计学意义; 免疫组化结果显示:与对照组比较,LPS组p38MAPK、ERK蛋白在胞质和胞核表达均明显增加,而UTI组较LPS组表达均明显减少; Western blot检测结果显示:与对照组比较,LPS组肺组织p-p38MAPK/ERK蛋白表达均明显增高,而UTI干预组蛋白表达均明显受抑制。结论 乌司他丁通过干预p38MAPK/ERK信号通路、抗氧化作用而减轻炎症反应,发挥对LPS致脓毒性大鼠ARDS的保护作用。
Abstract:
Aim To investigate whether ulinastatin has a beneficial effect on lipopolysaccharide(LPS)induced acute respiratory distress syndrome(ARDS)in rats,and to explore the possible underlying mechanisms.Methods Fifty-six Wistar rats were randomly assigned into control group, model group(LPS 6,12,24 h groups), ulinastatin group(UTI 6,12,24 h groups), with 8 in each group. ARDS rat model was reproduced by intraperitoneal injection of LPS(10 mg·kg-1), The rats in UTI groups were injected ulinastatin(20 000 u·kg-1), The rats in the control group received an equal volume of normal saline at the same time, rats in each group were sacrificed at 6,12,24 hours after LPS challenge. Plasma and lung tissue samples were collected, Histopathological evaluation, lung wet/dry(W/D)ratio, Tumor necrosis factor-a(TNF-α), Interleukin-18(IL-18),surfactant protein A(SPA), malondialdehyde(MDA),nitric oxide(NO)and superoxide dismutase(SOD)were analyzed. Immunohistochemical method was performed to detect the protein expression of p38MAPK and ERK. Western blot method was used to detect lung phosphorylated p38MAPK(p-p38MAPK)and pERK protein expression changes.Result In the control groups, lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group, ARDS characters such as extensive thickening of the alveolar wall, significant infiltration of inflammatory cells,demolished structure of pulmonary alveoli,and hemorrhage were found. In the all UTI treatment groups,these pathological changes in lung were markedly alleviated compared with those of LPS-induced ARDS group. Compared with control groups, lung W/D ratio,tumor necrosis factor-a(TNF-α),interleukin-18(IL-18)and surfactant protein A(SPA)in plasma,and lung MDA,NO levels in lung homogenates in the LPS group were increased significantly, while the lung SOD levels in the LPS group were decreased. Compared with the LPS group,lung W/D ratio,TNF-aIL-18 and(SPAin plasma,and lung MDA levels in lung homogenates in the UTI groups were decreased significantly, while the lung SOD levels in the UTI groups were increased. Immunohistochemistry showed that positive expressions of p38MAPK and ERK in cytoplasm and nucleus in the ulinastatin treatment groups were significantly lower than those in the model group. Western blot showed that compared with the control group, the p-p38MAPK and pERK protein expression in LPS group were significantly increased, and the ulinastatin could inhibit the protein expressions compared with model group.Conclusion Ulinastatin can significantly ameliorate the lung injury induced by LPS in rats via the intervention of p38MAPK and ERK signaling pathway and reducing inflammation and antioxidant effect.

参考文献/References:

[1] 乔 良,刘 志.按柏林新标准分析急诊脓毒症患者发生急性呼吸窘迫综合征的危险因素[J].中华危重病急救医学,2015,27(7):558-62. [1] Qiao L,Liu Z.Analysis of the risk factors of acute respiratory distress syndrome of Berlin new definition in patients with sepsis in emergency department[J]. Chinese Critical Care Medicine,2015; 27:558-62.
[2] Jonsson-Berling B M,Ohlsson K,Rosengren M.Radio immunological quantitation of the urinary trypsin inhibitor in normal blood and urine[J].Biol Chem HoPPe Seyler,1989,370(11):1157-61.
[3] 孟 莹,余常辉,李 婷,等.Toll样受体4在烟熏和脂多糖联合烟熏所致肺损伤大鼠中的表达及意义[J].中华医学杂志,2013,93(28):2230-4.
[3] Meng Y,Yu C H,Li T, et al.Expression and significance of Toll-like receptor-4 in rats lung established by passive smoking or associated with intratracheal instillation of lipopolysaccharide[J]. Chin Med J,2013,93:2230-4.
[4] Inoue K,Takano H,Smada A,et al.Urinary trypsin inhibitor protects against systemic inflammation induecd by lipopolysae charide[J].Mol Pharmacol,2005,67(3):673-80.
[5] Inoue K, Takano H.Urinary trypsin inhibitor as a therapeutic option for endotoxin-related inflammatory disorders[J].Expert Opin Investig Drugs,2010,19(4):513-20.
[6] Yu J B,Yao S L.Protective effects of hemin pretreatment combined with ulinastent on septic shock in rats[J].Chin Med J(Engl),2008,121(1):49-55.
[7] 陈 怿,童华生,张兴钦,等.乌司他丁减轻重症中暑大鼠肺的炎症和氧化损伤[J].中华急诊医学杂志,2014,23(8):866-70.
[7] Chen Y,Tong H S,Zhang X Q,et al. The study of ulinastatin to reduce pulmonary inflammation and oxidative injury in rats with severe heatstroke[J].Chinese Journal of Emergency Medicine,2014,23(8):866-70.
[8] Wang S Y, Li Z J, Wang X L,et al. Effect of ulinastatin on HMGB1 expression in rats with acute lung injury induced by sepsis[J]. Genet Mol Res,2015,14(2):4344-53.
[9] 朱桂军,孟志强,刘丽霞,等.DATS通过p38MAPK通路抑制LPS诱导的小鼠MH-S细胞TNF-α及IL-1β表达[J].中国药理学通报,2012,28(9):1303-7.
[9] Zhu G J,Meng Z Q,Liu L X,et al. Inhibition of LPS-induced TNF-α and IL-1β expression by DATS through p38 MAPK pathway in MH-S cell[J]. Chin Pharmacol Bull, 2012,28(9):1303-7.
[10] Eidt M V, Nunes F B, Pedrazza L, et al.Biochemical and inflammatory aspects in patients with severe sepsis and septic shock: The predictive role of IL-18 in mortality[J].Clin Chim Acta,2016,453:100-6
[11] Linder A, Russell J A. An exciting candidate therapy for sepsis:ulinastatin, a urinary protease inhibitor[J].Intensive Care Med,2014,40(8):1164-7.
[12] Rice T W,Bemard G R.Acute lung injury and the acute respiratory distress syndrome:challenges in clinieal trial design[J].Clin Chest Med,2006,27(4):733-54.
[13] 倪志宇,丛 斌,董 玫,等.八肽胆囊收缩素通过p38MAPK通路抑制LPS诱导的RAW264.7细胞IL-1β表达[J].中国药理学通报,2011,27(7):971-5.
[13] Ni Z Y,Cong B,Dong M,et al. CCK-8 inhibits LPS-induced IL-1β production in RAW264.7 cells by p38 MAPK pathway[J]. Chin Pharmacol Bull,2011,27(7):971-5.
[15] Di Paola Rl, Crisafulli C, Mazzon E, et al. Effect of PD98059,a selective MAPK3/MAPK1 inhibitor,on acute lung injury in mice[J].Int J Immunopathol Pharmacol,2009,22(4):937-50.
[16] Sawada K, Ariki S, Kojima T, et al. Pulmonary collectins protect macrophages against poreforming activity of Legionella pneumophila and suppress its intracellular growth[J]. J Biol Chem,2010,285(11):8434-8.

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备注/Memo

备注/Memo:
基金项目:内蒙古自治区自然基金项目(No 2013MS1128); 内蒙古自治区卫生与计划生育委员会基金(No 201302077)
作者简介:张利鹏(1980-),男,博士生,研究方向:脓毒症及ARDS的基础与临床,E-mail:zlp_boy2008@163.com
周丽华(1970-),女,博士,研究方向:脓毒症心肌损伤的信号通路研究,通讯作者,E-mail:rocboy2008@aliyun.com
更新日期/Last Update: 2016-09-15