5-脂氧合酶激活蛋白选择性抑制剂MK886对酒精性肝病小鼠模型的影响

杨淑娟; 郭雅玲; 付珊; 何英利; 赵英仁; 田臻

doi:10.3969/j.issn.1001-5256.2019.08.028

5-脂氧合酶激活蛋白选择性抑制剂MK886对酒精性肝病小鼠模型的影响

DOI: 10.3969/j.issn.1001-5256.2019.08.028

西安交通大学第一附属医院感染科
西安交通大学附属西安市第八医院肝病科
西安交通大学肝病研究所

基金项目:

国家自然科学基金(81800548)；

详细信息

中图分类号: R575;R-332
计量
- 文章访问数: 1357
- HTML全文浏览量: 53
- PDF下载量: 240
- 被引次数: 0
出版历程
- 收稿日期: 2019-03-19
- 出版日期: 2019-08-20

Role of the specific inhibitor of 5-lipoxygenase-activating protein MK886 in inhibiting alcoholic liver disease in mice

Department of Infectious Diseases, The First Affiliated Hospital of Xi'an Jiaotong University

Research funding:

摘要

摘要:
目的旨在探究5-脂氧合酶激活蛋白（FLAP）的选择性抑制剂MK886对小鼠酒精性肝病（ALD）的影响。方法 48只雄性昆明种小鼠随机分为4组,ALD组、ALD/MK886组给予Tipoe-Nanji酒精液体饲料,6周后酒精灌胃1次形成慢加急性ALD模型,对照组给予不含酒精的对照饲料,正常组给予普通饲料。小鼠开始摄入酒精2 d后,ALD/MK886组小鼠腹腔开始注射MK886（0. 01 mg/10 g,1次/d）。ALD组及ALD/MK886组酒精灌胃后9 h处死小鼠。检测血清中AST、ALT、乳酸脱氢酶（LDH）、TG及肝组织中TG和丙二醛（MDA）水平,肝组织行HE染色并进行病理评分,Western Blot法测定肝组织/Thp-1细胞中FLAP和5-脂氧合酶（5-LO）的表达水平,流式细胞法检测Thp-1细胞的凋亡水平。多组间比较采用单因素方差分析,方差齐时用LSD-t检验进行两两比较,方差不齐时采用Tamhane’T2进行检验。结果 ALD组、ALD/MK886组在造模第1周体质量减轻,随后逐渐增加,至造模结束时ALD组小鼠体质量和肝指数明显低于正常组和对照组（P值均<0...
- 肝疾病,酒精性 /
- 脂氧合酶 /
- 枯否细胞 /
- 细胞凋亡 /
- 小鼠,近交C57BL
Abstract:
Objective To investigate the effect of MK886, a specific inhibitor of 5-lipoxygenase-activating protein (FLAP) , on alcoholic liver disease (ALD) in mice. Methods A total of 48 male Kunming mice were randomly divided into ALD group, ALD/MK886 group, control group, and normal group. The mice in the ALD group and the ALD/MK886 group were fed with Tipoe-Nanji alcohol liquid diet and were given alcohol by gavage 6 weeks later to establish a model of acute ALD. The mice in the control group were given control diet without alcohol, and those in the normal group were given normal diet. After 2 days of alcohol intake, the mice in the ALD/MK886 group were given intraperitoneally injected MK886 (0. 01 mg/10 g) once a day. The mice were sacrificed at 9 hours after the administration of alcohol by gavage. Serum levels of aspartate aminotransferase (AST) , alanine aminotransferase (ALT) , lactate dehydrogenase (LDH) , and triglyceride (TG) and the levels of TG and malondialdehyde (MDA) in liver tissue were measured. HE staining was performed for liver tissue and pathological score was determined. Western blot was used to measure the expression of FLAP and 5-lipoxygenase (5-LO) in liver tissue and Thp-1 cells, and flow cytometry was used to measure the apoptosis of Thp-1 cells. One-way ANOVA was used for comparison between groups; the least significant difference t-test was used for comparison of data with homogeneity of variance, and the Tamhane's T2 test was used for comparison of data with heterogeneity of variance. Results The ALD group and the ALD/MK886 group had a reduction in body weight within the first week of modeling, followed by a gradual increase; at the end of modeling, the ALD group had significantly lower bodyweight and liver index than the normal group and the control group (all P < 0. 05) , and the ALD/MK886 group had significantly higher body weight and liver index than the ALD group (both P < 0. 05) . Compared with the normal group and the control group, the ALD group had significant increases in the levels of AST, ALT, LDH, and TG in serum and the levels of TG and MDA in liver tissue (all P < 0. 05) , and compared with the ALD group, the ALD/MK886 group had significant reductions in the above indices (all P < 0. 05) . The ALD group had a significantly higher score of hepatic steatosis than the normal group and the control group (both P < 0. 05) , and compared with the ALD group, the ALD/MK886 group had a significant reduction in this score (P < 0. 05) . Compared with the normal group and the control group, the ALD group had significant increases in the expression of 5-LO and FLAP in the liver (all P < 0. 05) , and the ALD/MK886 group had significant reductions compared with the ALD group (P < 0. 05) . Lipopolysaccharide (LPS) upregulated the expression of FLAP and 5-LO in Thp-1 cells, and MK886 alleviated such effect of LPS (all P < 0. 05) ; MK886 promoted the apoptosis of Thp-1 cells, and LPS alleviated the effect of MK886 in promoting cell apoptosis (all P < 0. 01) . Conclusion MK886 can inhibit the expression of 5-LO, induce the apoptosis of Kupffer cells, and thus exert a therapeutic effect on ALD in mice.
- liver diseases, alcoholic /
- lipoxygenase /
- kupffer cell /
- apoptosis /
- mice, inbred C57BL

HTML全文

参考文献(21)

[1] OSNA NA, DONOHUE TM Jr, KHARBANDA KK. Alcoholic liver disease:Pathogenesis and current management[J]. Alcohol Res, 2017, 38 (2) :147-161.

[2] VASSALLO GA, TARLI C, RANDO MM, et a. Liver transplantation in patients with alcoholic liver disease:A retrospective study[J]. Alcohol Alcohol, 2018, 53 (2) :151-156.

[3] National Workshop on Fatty Liver and Alcoholic Liver Disease, Chinese Society of Hepatology, Chinese Medical Association;Fatty Liver Expert Committee, Chinese Medical Doctor Association. Guidelines of prevention and treatment for alcoholic liver disease:A 2018 update[J]. J Clin Hepatol, 2018, 34 (5) :939-946. (in Chinese) 中华医学会肝病学分会脂肪肝和酒精性肝病学组, 中国医师协会脂肪性肝病专家委员会.酒精性肝病防治指南 (2018年更新版) [J].临床肝胆病杂志, 2018, 34 (5) :939-946.

[4] STICKEL F, DATZ C, HAMPE J, et al. Pathophysiology and management of alcoholic liver disease:Update 2016[J]. Gut Liver, 2017, 11 (2) :173-188.

[5] KOLIOS G, VALATAS V, KOUROUMALIS E. Role of Kupffer cells in the pathogenesis of liver disease[J]. World J Gastroenterol, 2006, 12 (46) :7413-7420.

[6] BALA S, CSAK T, KODYS K, et al. Alcohol-induced miR-155 and HDAC11 inhibit negative regulators of the TLR4 pathway and lead to increased LPS responsiveness of Kupffer cells in alcoholic liver disease[J]. J Leukoc Biol, 2017, 102 (2) :487-498.

[7] PARK JK, SHAO M, KIM MY, et al. An endoplasmic reticulum protein, Nogo-B, facilitates alcoholic liver disease through regulation of kupffer cell polarization[J]. Hepatology, 2017, 65 (5) :1720-1734.

[8] SORGI CA, ZARINI S, MARTIN SA, et al. Dormant 5-lipoxygenase in inflammatory macrophages is triggered by exogenous arachidonic acid[J]. Sci Rep, 2017, 7 (1) :10981.

[9] BRUNO F, ERRICO S, PACE S, et al. Structural insight into the optimization of ethyl 5-hydroxybenzo[g]indol-3-carboxylates and their bioisosteric analogues as 5-LO/m-PGES-1 dual inhibitors able to suppress inflammation[J]. Eur J Med Chem, 2018, 155:946-960.

[10] TITOS E, CLRIA J, PLANAGUMA, et al. Inhibition of 5-lipoxygenase-activating protein abrogates experimental liver injury:Role of Kupffer cells[J]. J Leukoc Biol, 2005, 78 (4) :871-878.

[11] LEO LM, ALMEIDA-CORRA S, CANETTI CA, et al. Agedependent relevance of endogenous 5-lipoxygenase derivatives in anxiety-like behavior in mice[J]. PLo S One, 2014, 9 (1) :e85009.

[12] SUMMA KC, VOIGT RM, FORSYTH CB, et al. Disruption of the circadian clock in mice increases intestinal permeability and promotes alcohol-induced hepatic pathology and inflammation[J]. PLo S One, 2013, 8 (6) :e67102.

[13] DESMET VJ, KNODELL RG, ISHAK KG, et al. Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis[Hepatology 1981; 1:431-435][J]. J Hepatol, 2003, 38 (4) :382-386.

[14] MASARONE M, ROSATO V, DALLIO M, et al. Epidemiology and natural history of alcoholic liver disease[J]. Rev Recent Clin Trials, 2016, 11 (3) :167-174.

[15] LOUVET A, MATHURIN P. Alcoholic liver disease:Mechanisms of injury and targeted treatment[J]. Nat Rev Gastroenterol Hepatol, 2015, 12 (4) :231-242.

[16] SZABO G, PETRASEK J. Gut-liver axis and sterile signals in the development of alcoholic liver disease[J]. Alcohol Alcohol, 2017, 52 (4) :414-424.

[17] LIN XX, CHEN D, ZHAO Q, et al. Genetic reasearch on the incidence of alcoholic liver disease[J]. Chin J Clin Pharmacol Ther, 2017, 22 (11) :1309-1314. (in Chinese) 林秀贤, 陈丹, 赵青, 等.影响酒精性肝病发病的遗传学研究进展[J].中国临床药理学与治疗学, 2017, 22 (11) :1309-1314.

[18] NADARAJAN K, BALARAM P, KHOO BY. MK886 inhibits the pioglitazone-induced anti-invasion of MDA-MB-231 cells is associated with PPARα/γ, FGF4 and 5LOX[J]. Cytotechnology, 2016, 68 (5) :1771-1787.

[19] DAGLAR GO, KAMA NA, ATLI M, et al. Effect of 5-lipoxygenase inhibition on Kupffer cell clearance capacity in obstructive jaundiced rats[J]. J Surg Res, 2001, 96 (2) :158-162.

[20] ZHOU H, YU M, ZHAO J, et al. IRAKM-Mincle axis links cell death to inflammation:Pathophysiological implications for chronic alcoholic liver disease[J]. Hepatology, 2016, 64 (6) :1978-1993.

[21] KOOP DR, KLOPFENSTEIN B, IIMURO Y, et al. Gadolinium chloride blocks alcohol-dependent liver toxicity in rats treated chronically with intragastric alcohol despite the induction of CYP2E1[J]. Mol Pharmacol, 1997, 51 (6) :944-950.

施引文献

资源附件(0)

访问统计