AMPK参与调节大鼠脂肪肝相关性肝癌前病变的形成

doi:10.3969/j.issn.1006-7795.2016.02.019

首都医科大学学报 ›› 2016, Vol. 37 ›› Issue (2): 208-213.doi: 10.3969/j.issn.1006-7795.2016.02.019

AMPK参与调节大鼠脂肪肝相关性肝癌前病变的形成

王芸姣, 韩文祺, 李若菲, 杜尊赎, 王学江, 江瑛

首都医科大学基础医学院生理学与病理生理学系病理生理学教研室, 北京 100069

收稿日期:2016-01-12 出版日期:2016-04-21 发布日期:2016-04-14
通讯作者: 江瑛 E-mail:jiangy@ccmu.edu.cn
基金资助:
国家自然科学基金(31171032)

Adenosine monophosphate-activated protein kinase involved in early hepatocarcinogenesis associated with nonalcoholic steatohepatitis in rats

Wang Yunjiao, Han Wenqi, Li Ruofei, Du Zunshu, Wang Xuejiang, Jiang Ying

Division of Pathophysiology, Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China

Received:2016-01-12 Online:2016-04-21 Published:2016-04-14
Supported by:
This study was supported by National Natural Science Foundation of China(31171032).

摘要/Abstract

摘要： 目的探讨单磷酸腺苷活化蛋白激酶(adenosine monophosphate-activated protein kinase,AMPK)在低剂量二乙基亚硝胺(diethylnitrosamine, DEN)合并高脂饮食诱导的大鼠肝癌前病变发生中的作用及其机制。方法体内实验采用腹腔注射DEN(30 mg/kg)合并高脂饮食饲喂大鼠16周诱导肝癌前病变模型,通过HE染色、Western blotting、Real-time PCR、免疫组织化学等方法观察谷胱甘肽S转移酶(glutathione S-transferase-π,GST-π)、固醇调节元件结合蛋白1c(sterol regulatory element binding protein-1c, SREBP-1c)、脂肪酸合成酶(fatty acid synthase,FAS)、乙酰辅酶A羧化酶(acetyl-CoA carboxylase,ACC)、硬脂酰辅酶A去饱和酶1(stearoyl-CoA desaturase 1,SCD1)及AMPK、p-AMPK的表达变化;体外实验观察AMPK对棕榈酸(palmitic acid, PA)诱导的大鼠H4IIE细胞脂质代谢的影响。结果与单纯DEN处理组比较,DEN+高脂组大鼠肝细胞脂肪变性、气球样变、伴有炎性细胞浸润及小灶性坏死;GST-π表达水平增高;三酰甘油(triglyceride,TG)及SREBP-1c、FAS、ACC、SCD1表达水平上升;p-AMPK水平下降。AMPK通过抑制SREBP-1c的表达水平降低棕榈酸诱导的H4IIE细胞内脂质合成。结论 AMPK可能通过抑制SREBP-1c的表达水平参与大鼠肝癌前病变的形成。

关键词: 二乙基亚硝胺, 非酒精性脂肪性肝炎, 肝癌前病变, 单磷酸腺苷活化蛋白激酶AMPK, 固醇调节元件结合蛋白1c(SREBP-1c)

Abstract: Objective To explore the role of adenosine monophosphate-activated protein kinase(AMPK) in the development of precancerosis induced by a high-fat diet and diethylnitrosamine and its molecular mechanism.Methods The low dose of diethylnitrosamine(DEN, 30mg/kg, ip) and high fat diet(16 weeks)induced liver precancerosis of rat model in vivo was established to observe the pathological changes of rat livers by HE staining.The expression levels of glutathione S-transferase-π(GST-π), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), stearoyl-CoA desaturase 1(SCD1), AMPK, p-AMPK were detected by Western blotting, Real-time PCR and immunohistochemistry. The rat cells H4IIE were disposed by palmitic acid(PA) to mimic lipid overload in hepatocarcinoma cells and evaluate the role of AMPK in modulating fat metabolism of hepatocarcinoma.Results We found lipid droplets overload evidently in hepatocytes of rats in DEN+HFD group compared with DEN group, some of which behaving ballooning change, necrosis and inflammatory; meanwhile, GST-π and triglyceride(TG) expression were both elevated significantly in the livers of rats treated with DEN+HFD group when compared with that of DEN group, indicating the precancerosis of rat model induced by NASH was successful.Similarly, the expression of SREBP-1c and its target genes FAS, ACC, SCD1 were all elevated statistically. Furthermore, p-AMPK level was decreased. The in vitro model indicated that TG, SREBP-1c, FAS, ACC, SCD1 were all increased in PA treated cells, while treating cells with AICAR, activator of AMPK, can reverse the effect of PA. Conclusion AMPK is involved in liver precancerosis induced by high-fat diet and diethylnitrosamine, and this process may happen through the inhibition of SREBP-1c.

Key words: diethylnitrosamine, non-alcoholic steatohepatitis, liver precancerosis, adenosine monophosphate-activated protein kinase, sterol regulatory element binding protein-1c

中图分类号:

R575.5

王芸姣, 韩文祺, 李若菲, 杜尊赎, 王学江, 江瑛. AMPK参与调节大鼠脂肪肝相关性肝癌前病变的形成[J]. 首都医科大学学报, 2016, 37(2): 208-213.

Wang Yunjiao, Han Wenqi, Li Ruofei, Du Zunshu, Wang Xuejiang, Jiang Ying. Adenosine monophosphate-activated protein kinase involved in early hepatocarcinogenesis associated with nonalcoholic steatohepatitis in rats[J]. Journal of Capital Medical University, 2016, 37(2): 208-213.

参考文献

[1] Rinella M E. Nonalcoholic fatty liver disease:a systematic review[J]. JAMA, 2015,313(22):2263-2273.
[2] Farrell G C, Larter C Z. Nonalcoholic fatty liver disease:from steatosis to cirrhosis[J]. Hepatology, 2006,43(2 Suppl 1):S99-S112.
[3] Saran U, Humar B, Kolly P, et al. Hepatocellular carcinoma and lifestyles[J]. J Hepatol, 2016,64(1):203-214.
[4] Guzman G, Brunt E M, Petrovic L M, et al. Does nonalcoholic fatty liver disease predispose patients to hepatocellular carcinoma in the absence of cirrhosis?[J]. Arch Pathol Lab Med, 2008,132(11):1761-1766.
[5] Park E J, Lee J H, Yu G Y, et al. Dietary and genetic obesity promote liver inflammation and tumorigenesis by enhancing IL-6 and TNF expression[J]. Cell, 2010,140(2):197-208.
[6] Wang Y, Ausman L M, Greenberg A S, et al. Nonalcoholic steatohepatitis induced by a high-fat diet promotes diethylnitrosamine initiated early hepatocarcinogenesis in rats[J]. Int J Cancer,2009,124(3):540-546.
[7] Park K G, Min A K, Koh E H, et al. Alpha-lipoic acid decreases hepatic lipogenesis through adenosine monophosphate-activated protein kinase (AMPK)-dependent and AMPK-independent pathways[J]. Hepatology, 2008,48(5):1477-1486.
[8] Zheng L, Yang W, Wu F, et al. Prognostic significance of AMPK activation and therapeutic effects of metformin in hepatocellular carcinoma[J]. Clin Cancer Res, 2013,19(19):5372-5380.
[9] Sato K, Kitahara A, Satoh K, et al. The placental form of glutathione S-transferase as a new marker protein for preneoplasia in rat chemical hepatocarcinogenesis[J]. Gan, 1984,75(3):199-202.
[10] Dyson J, Jaques B, Chattopadyhay D, et al. Hepatocellular cancer:the impact of obesity, type 2 diabetes and a multidisciplinary team[J]. J Hepatol, 2014,60(1):110-117.
[11] Caldwell S H, Crespo D M, Kang H S, et al. Obesity and hepatocellular carcinoma[J]. Gastroenterology, 2004,127(5 Suppl 1):S97-103.
[12] Oh S W, Yoon Y S, Shin S A. Effects of excess weight on cancer incidences depending on cancer sites and histologic findings among men:Korea National Health Insurance Corporation Study[J]. J Clin Oncol, 2005,23(21):4742-4754.
[13] 中华医学会肝病学分会脂肪肝和酒精性肝病学组.非酒精性脂肪性肝病诊疗指南(2010年修订版)[J]. 胃肠病学和肝病学杂志,2010,19(6):483-487.
[14] Adams L A, Lymp J F, St Sauver J, et al. The natural history of nonalcoholic fatty liver disease:a population-based cohort study[J]. Gastroenterology, 2005,129(1):113-121.
[15] Szlosarek P W, Balkwill F R. Tumour necrosis factor alpha:a potential target for the therapy of solid tumours[J]. Lancet Oncol, 2003,4(9):565-573.
[16] Novikova D S, Garabadzhiu A V, Melino G, et al. AMP-activated protein kinase:structure, function, and role in pathological processes[J]. Biochemistry (Mosc), 2015,80(2):127-144.
[17] Dasgupta B, Chhipa R R. Evolving Lessons on the Complex Role of AMPK in Normal Physiology and Cancer[J]. Trends Pharmacol Sci,2016,37(3):192-206.
[18] Li N S, Zou J R, Lin H, et al. LKB1/AMPK inhibits TGF-β1 production and the TGF-β signaling pathway in breast cancer cells[J]. Tumour Biol, 2015.[Epub ahead of print]
[19] Wu T, Wang M C, Jing L, et al. Autophagy facilitates lung adenocarcinoma resistance to cisplatin treatment by activation of AMPK/mTOR signaling pathway[J]. Drug Des Devel Ther, 2015,9:6421-6431.
[20] Cheng J, Huang T, Li Y, et al. AMP-activated protein kinase suppresses the in vitro and in vivo proliferation of hepatocellular carcinoma[J]. PLoS One, 2014,9(4):e93256.
[21] Jeon T I, Osborne T F. SREBPs:metabolic integrators in physiology and metabolism[J]. Trends Endocrinol Metab, 2012,23(2):65-72.
[22] Li C, Yang W, Zhang J, et al. SREBP-1 has a prognostic role and contributes to invasion and metastasis in human hepatocellular carcinoma[J]. Int J Mol Sci, 2014,15(5):7124-7138.

AMPK参与调节大鼠脂肪肝相关性肝癌前病变的形成

Adenosine monophosphate-activated protein kinase involved in early hepatocarcinogenesis associated with nonalcoholic steatohepatitis in rats

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 4

编辑推荐

Metrics

本文评价

[1]	覃岭;王宇童. 过表达apoA-Ⅰ对BEL-7402细胞内脂质堆积的影响[J]. 首都医科大学学报, 2012, 33(2): 198-204.
[2]	武彦宁;蔡照华_;孙海梅;尚宏伟;郝刚;张立新;孙琳;张华;丁惠国. 非酒精性脂肪性肝炎大鼠肝脏内源性H₂S合成减少[J]. 首都医科大学学报, 2010, 31(3): 287-292.
[3]	崔焱;张莉;贾继东. 己酮可可碱治疗非酒精性脂肪性肝炎的实验研究[J]. 首都医科大学学报, 2009, 30(3): 317-320.
[4]	任燕龙;孙海梅;曾晓蓓;尚宏伟;张华;安威. 己酮可可碱对非酒精性脂肪性肝炎大鼠肝组织MMP-9、TGFβ1表达的影响[J]. 首都医科大学学报, 2008, 29(4): 463-466.