细胞自噬在D-氨基半乳糖/脂多糖诱导的小鼠急性肝损伤模型中的保护作用及机制

时红波; 时红林; 张向颖; 陈德喜; 段钟平; 任锋

doi:10.3969/j.issn.1001-5256.2017.02.027

细胞自噬在D-氨基半乳糖/脂多糖诱导的小鼠急性肝损伤模型中的保护作用及机制

DOI: 10.3969/j.issn.1001-5256.2017.02.027

首都医科大学附属北京佑安医院北京市肝病研究所

基金项目:

国家自然科学基金(81300349,81270532)；北京市自然科学基金(7162085,7144216)；北京市科技新星计划(Z131107000413016)；首都临床特色研究(Z161100000516113)；北京市医院管理局临床医学发展专项经费资助(XM201308)；国家临床重点专科建设项目(WJWYA-2014-002)；科研基地建设-重大传染病防治协同创新中心(115215)；北京卫生系统高技术人才培养项目资助(2014-3-090,2013-3-075)；北京市医院管理局登峰计划专项经费资助(DFL20151601)；

详细信息

中图分类号: R575;R-332
计量
- 文章访问数: 1717
- HTML全文浏览量: 82
- PDF下载量: 443
- 被引次数: 0
出版历程
- 收稿日期: 2016-10-17
- 出版日期: 2017-02-20

Protective effect of autophagy in mice with acute liver injury induced by D- galactosamine / lipopolysaccharide and related mechanisms

Beijing You An Hospital,Capital Medical University

Research funding:

摘要

摘要:
目的利用D-氨基半乳糖（D-Gal N）/脂多糖（LPS）诱导的小鼠急性肝损伤模型研究细胞自噬在急性肝损伤中的作用及机制。方法以C57BL/6小鼠为研究对象,腹腔注射D-Gal N/LPS建立小鼠急性肝损伤模型。动物实验分组:对照组、DGal N/LPS组、雷帕霉素+D-Gal N/LPS组、三甲基腺嘌呤（3-MA）+D-Gal N/LPS组、Atg7 siRNA+D-Gal N/LPS组。观察不同分组中小鼠的生存情况,观察肝组织病理变化评价肝损伤情况,全自动生化分析仪检测血清ALT、AST水平,实时荧光定量PCR检测肝组织中TNFα和IL-6基因表达,荧光显微镜观察肝组织中肝细胞凋亡情况。多样本组间比较采用One-way ANOVA分析,进一步两两比较,方差齐时采用LSD-t检验,方差不齐时采用Games-Howell法。结果与D-Gal N/LPS组相比,应用自噬激活剂雷帕霉素干预后,小鼠生存率明显升高（80%vs 40%）,肝脏出血、炎症和坏死明显减轻,肝细胞凋亡明显减少,血清ALT、AST水平明显降低[ALT:（427.4±195.5）U/L vs（977.7±247.3）U/...
- 自噬 /
- 肝功能衰竭,急性 /
- 己糖胺酶类 /
- 脂多糖类 /
- 小鼠,近交C57BL
Abstract:
Objective To investigate the role and mechanism of autophagy in acute liver injury induced by D- galactosamine / lipopolysaccharide( D- Gal N / LPS) in mice.Methods C57 BL /6 mice were used to establish a mouse model of acute liver injury using intraperitoneally injected D- Gal N / LPS.In this animal experiment,the mice were divided into control group,D- Gal N / LPS group,rapamycin + D-Gal N / LPS group,3- MA + D- Gal N / LPS group,and Atg7 siRNA + D- Gal N / LPS group.The survival of the mice was observed,and liver pathological changes were observed to analyze liver injury.An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase( ALT) and aspartate aminotransferase( AST),quantitative real- time PCR was performed to measure the mRNA expression of tumor necrosis factorα( TNFα) and interleukin- 6( IL- 6),and a fluorescence microscope was used to observe the apoptosis of hepatocytes.A One- way ANOVA was used for comparison between multiple groups;the least significant difference t- test was used for homogeneity of variance and the Games- Howell method was used for heterogeneity of variance.Results Compared with the D- Gal N / LPS group,the rapamycin + D- Gal N / LPS group had a significant increase in survival rate( 80% vs 40%),significantly reduced liver hemorrhage,inflammation,and necrosis and apoptosis of hepatocytes,and significant reductions in serum levels of ALT( 427.4 ± 195.5 U / L vs977.7 ± 247.3 U / L,P = 0.002) and AST( 378.2 ± 169.7 U / L vs 1100.0 ± 438.0 U / L,P = 0.004),as well as significant reductions in the mRNA expression of TNFα( 0.288 ± 0.010 vs 1.136 ± 0.267,P = 0.003) and IL- 6( 0.272 ± 0.061 vs 0.869 ± 0.317,P =0.010).Compared with the D- Gal N / LPS group,the 3- MA + D- Gal N / LPS group and Atg7 siRNA + D- Gal N / LPS group had significant reductions in survival rate( 0% /10% vs 40%),significantly aggravated liver hemorrhage,inflammation,and necrosis and apoptosis of hepatocytes,and significant increases in serum levels of ALT( 1836.0 ± 560.5 U / L and 1654.0 ± 627.6 U / L vs 977.7 ± 247.3 U / L,P =0.006 and 0.034) and AST( 1948.0 ± 645.5 U / L and 1804.0 ± 492.6 U / L vs 1100.0 ± 438.0 U / L,P = 0.029 and 0.033),as well as significant increases in the expression of TNFα in liver tissue( 2.026 ± 0.342 and 1.994 ± 0.286 vs 1.136 ± 0.267,P = 0.006 and0.005).Conclusion In the mouse model of acute liver injury induced by D- Gal N / LPS,autophagy has an important protective effect,and its regulating mechanism may be associated with the inhibitory effect on inflammatory factors and apoptosis of hepatocytes.
- autophagy /
- liver failure /
- acute /
- hexosaminidases /
- lipopolysaccharides

HTML全文

参考文献(22)

[1]KHOURY T,RMEILEH AA,YOSHA L,et al.Drug induced liver injury:review with a focus on genetic factors,tissue diagnosis,and treatment options[J].J Clin Transl Hepatol,2015,3(2):99-108.

[2]OH IS,PARK SH.Immune-mediated liver injury in hepatitis B virus infection[J].Immune Netw,2015,15(4):191-198.

[3] Liver Failure and Artificial Liver Group,Chinese Society of Infectious Diseases,Chinese Medical Association;Severe Liver Diseases and Artificial Liver Group,Chinese Society of Hepatology,Chinese Medical Association.Diagnostic and treatment guidelines for liver failure[J].Chin J Hepatol,2013,21(3):177-183.(in Chinese)中华医学会感染病学分会肝衰竭与人工肝学组,中华医学会肝病学分会重型肝病与人工肝学组.肝衰竭诊疗指南(2012年版)[J].中华肝脏病杂志,2013,21(3):177-183.

[4] WEI LL,ZHANG XY,ZHANG L,et al.The expression of autophagy in progression of acute liver failure induced by D-galactosamine/lipopolysaccharide in mice[J].Chin J Hepatol,24(5):120-125.(in Chinese)魏琳琳,张向颖,张莉,等.细胞自噬在D-氨基半乳糖/脂多糖诱导小鼠急性肝衰竭中的表达[J].中华肝脏病杂志,2016,24(5):120-125.

[5]WANG K,DAMJANOV I,WAN YJ.The protective role of pregnane Xreceptor in lipopolysaccharide/D-galactosamine-induced acute liver injury[J].Lab Invest,2010,90(2):257-265.

[6]BETH L,NOBORU M,HERBERT W.Autophagy in immunity and inflammation[J].Nature,2011,469(7330):323-335.

[7]CHATTERJEE C,SPARKS DL.Hepatic lipase release is inhibited by a purinergic induction of autophagy[J].Cell Physiol Biochem,2014,33(4):883-894.

[8]SCHNEIDER JL,CUERVO AM.Liver autophagy:much more than just taking out the trash[J].Nat Rev Gastroenterol Hepatol,2014,11(3):187-200.

[9]CUI YL,QI C,WANG CF,et al.Role of autophapy in antigen recognition,processing,and presentation in the body's immune defense[J].Chin J Immunol,2015,31(11):1565-1568.(in Chinese)崔玉琳,齐翀,王春凤,等.细胞自噬在机体免疫防御中识别、加工、递呈抗原的作用[J].中国免疫学杂志,2015,31(11):1565-1568.

[10] NOBORU M,BETH L,ANA MC,et al.Autophagy fights disease through cellular self-digestion[J].Nature,2008,451(7182):1069-1075.

[11]KOMATSU M,KUROKAWA H,WAGURI S,et al.The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1[J].Nature Cell Biol,2010,12(3):213-223.

[12]LEVINE B,KROEMER G.Autophagy in the pathogenesis of disease[J].Cell,2008,132(1):27-42.

[13]CHANG CP,LEI HY.Autophagy induction in T cell-independent acute hepatitis induced by concanavalin A in SCID/NOD mice[J].Int J Immunopathol Pharmacol,2008,21(4):817-826.

[14]DIAO W,JIN F,WANG B,et al.The protective role of myeloidderived suppressor cells in concanavalin A-induced hepatic injury[J].Protein Cell,2014,5(9):714-724.

[15]RYU KH,KIM SY,KIM YR,et al.Tonsil-derived mesenchymal stem cells alleviate concanavalin A-induced acute liver injury[J].Exp Cell Res,2014,326(1):143-154.

[16]NI HM,BOCKUS A,BOGGESS N,et al.Activation of autophagy protects against acetaminophen-induced hepatotoxicity[J].Hepatology,2012,55(1):222-232.

[17]NI HM,WILLIAMS JA,JAESCHKE H,et al.Zonated induction of autophagy and mitochondrial spheroids limits acetaminophen-induced necrosis in the liver[J].Redox Biol,2013,1:427-432.

[18]NI HM,BOGGESS N,McG ILL MR,et al.Liver-specific loss of Atg5 causes persistent activation of Nrf2 and protects against acetaminophen-induced liver injury[J].Toxicol Sci,2012,127(2):438-450.

[19]SUN HY,WANG XQ,SHI HB,et al.Ganshuang granules protect mouse liver from chronic injury induced by CCl4via autophagy[J].J Clin Hepatol,2015,31(7):1114-1119.(in Chinese)孙海青,王小琪,时红波,等.肝爽颗粒对CCl4诱导的慢性肝损伤小鼠模型和肝损伤细胞模型的保护作用[J].临床肝胆病杂志,2015,31(7):1114-1119.

[20]TOSHIMA T,SHIRABE K,FUKUHARA T,et al.Suppression of autophagy during liver regeneration impairs energy charge and hepatocyte senescence in Mice[J].Hepatology,2014,60(1):290-300.

[21]REN F,ZHANG L,ZHANG X,et al.Inhibition of glycogen synthase kinase 3βpromotes autophagy to protect mice from acute liver failure mediated by peroxisome proliferator-activated receptorα[J].Cell Death Dis,2016,7:e2151.

[22]JIAO M,REN F,ZHOU L,et al.Peroxisome proliferator-activated receptorαactivation attenuates the inflammatory response to protect the liver from acute failure by promoting the autophagy pathway[J].Cell Death Dis,2014,5:e1397.

施引文献

资源附件(0)

访问统计