Regeneration and structural changes of mouse cirrhotic liver after partial hepatectomy

doi:10.3969/j.issn.2095-4344.2013.53.016

Abstract

Abstract:

BACKGROUND: Liver possesses a powerful regenerative capacity. It is reported that the most patients suffering from hepatocelluler carcinoma are accompanied with liver cirrhosis. The liver weight can be gradually restored after tumor resection, which indicated a regenerative potential of cirrhosis liver.

OBJECTIVE:To try to use partial hepatectomy to induce proliferation of cells with regeneration capacity in cirrhotic liver of mice, and to observe the structural changes of regenerated liver.

METHODS: The liver cirrhosis model of mouse was produced by injecting CCl₄ liquid subcutaneously. The mice with liver cirrhosis were subjected to partial hepatectomy. The residual liver was dissected and weighted at 1, 3, 5, 7, 10, 14 and 21 days after partial hepatectomy. The structural changes of regenerating livers were compared after partial hepatectomy at various time points. Liver regeneration rate was evaluated. Immunohistochemical method was used to analyze desmin and alpha-smooth muscle actin expression in the hepatic satellite cells. The 5-bromo-2-deoxyuridine incorporation labeling method was used to exhibit and localize the regenerating cells.

RESULTS AND CONCLUSION:The liver regeneration rates of the mice with liver cirrhosis at the 1, 3, 5, 7, 14, 21 days after the partial hepatectomy were 6.58%, 22.03%, 21.39%, 29.05%, 45.22%, and 50.98% respectively. The results of hematoxylin-eosin staining as well as Masson staining showed the pathological early cirrhosis features of the liver after CCl₄ treatment for 8 weeks and the structure of residual livers improved gradually after partial hepatectomy. Immunohistochemical results demonstrated that the expression of desmin in regenerating liver was decreased at 1 day after partial hepatectomy, but showed an uptrend from 3 to 14 days and decreased at 21 days. The expression of alpha-smooth muscle actin decreased gradually at every time point after partialhepatectomy. The 5-bromo-2- deoxyuridine-positive nucleus localized in hepatocytes at 1-7 days and mainly localized in hepatic sinusoid endothelium and intracavity at 14-21 days after partial hepatectomy. The regenerative reaction of mouse cirrhotic liver appeared at 3 days after partial hepatectomy and the structure of normal regenerating liver will be improved and restored gradually.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: liver cirrhosis, partial hepatectomy, liver regeneration, desmin, actins

CLC Number:

R318

Zheng Xiao-jun, Luo Qian-xin, Chen Qian-qi, Xiong Xi-feng, Liu Yu-rong, Ma Ning-fang. Regeneration and structural changes of mouse cirrhotic liver after partial hepatectomy[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(53): 9196-9202.

Figures/Tables 6

References

[1] 范婷婷,谢渭芬.肝纤维化和肝硬化治疗进展[J].国际消化病杂志, 2010,30(6): 349-351.

[2] Schuppan D, Kim YO. Evolving therapies for liver fibrosis. J Clin Invest. 2013;123(5):1887-1901.

[3] 汤钊猷.肝癌研究进展[J].中国肿瘤,2001,10(1): 37-40.

[4] 吴孟超,陈汉,沈锋. 原发性肝癌的外科治疗一附5524例报告[J].中华外科杂志,2001,39(1):25-28.

[5] Nagasue N, Kohno H, Chang YC, et al. Liver resection for hepatocelluar carcinoma. Ann Surg. 1993; 217(4): 375-384.

[6] Moser MA,Kneteman NM,Minuk GY. Research towards safer resection of the cirrhotic liver. HBP Surg. 2000;11: 285-297.

[7] Bravo E, D'Amore E, Ciaffoni F, et al. Evaluation of the spontaneous reversibility of carbon tetrachloride-induced liver cirrhosis in rabbits. Lab Anim. 2012;46(2):122-128.

[8] Mederacke I.Liver fibrosis-mouse models and relevance in human liver diseases. Z Gastroenterol. 2013;51(1):55-62.

[9] Higgins GM, Anderson RM. Experimental pathology of the liver. I. Restoration of the liver of the white rat following partial surgical removal. Arch Pathol Lab Med. 1931;12:186-202.

[10] Michalopoulos GK. Liver regeneration.J Cell Physiol. 2007; 213(2):286-300.

[11] 陈栋,吴力群,曹景玉,等.大鼠肝切除术后肝损伤程度与肝再生状态的动态对比研究[J].中华肝胆外科杂志,2002,8(6):354-357.

[12] Jones LD, Nielsen MK, Britton RA. Genetic variation in liver mass, body mass, and liver: body mass in mice. J Anim Sci. 1992;70: 2999-3006.

[13] Tan X, Behari J, Cieply B, et al. Conditional deletion of beta-catenin reveals its role in liver growth and regeneration. Gastroenterology. 2006; 131: 1561-1572.

[14] Inderbitzin D, Gass M, Beldi G, et al. Magnetic resonance imaging provides accurate and precise volume determination of the regenerating mouse liver. J Gastrointest Surg. 2004; 8: 806-811.

[15] Kumar S, Zou Y, Bao Q, et al. Proteomic analysis of immediate-early response plasma proteins after 70% and 90% partial hepatectomy. Hepatol Res. 2012. doi: 10.1111/hepr.12030. [Epub ahead of print]

[16] Hayashi H, Nagaki M, Imose M, et al. Normal liver regeneration and liver cell apoptosis after partial hepatectomy in tumor necrosis factor-alpha-deficient mice. Liver Int. 2005; 25:162-170.

[17] Fujita J, Marino MW, Wada H, et al. Effect of TNF gene depletion on liver regeneration after partial hepatectomy in mice. Surgery. 2001;129: 48-54.

[18] 方芳,徐丽萍,董沛杰,等.不同性别小鼠肝大部切除术后肝再生能力的比较[J].中国组织工程研究与临床康复,2010,14(53): 9909-9912.

[19] Selzner M, Clavian PA. Failure of regeneration of the steatotic rat liver: disruption at two different levels in the regeneration pathway. Hepatology. 2000;31: 35-42.

[20] Martins PN, Theruvath TP, Neuhaus P. Rodent models of partial hepatectomies. Liver Int.2008;28:3-11.

[21] Safadi R,Friedman SL.Hepatic fibrosis-role of hepatic stellate cell activation.Med Gen Med. 2002;4(3):27-32.

[22] Benyon RC,Iredale JP. Is liver fibrosis reversible? Gut. 2000; 46(4):443-446.

[23] Reeves HL,Friedman SL. Activation of hepatic stellate cells- a key issue in liver fibrosis. Front Biosci. 2002;7:808-826.

[24] 熊喜峰,何伟业,谢湘梅,等.β-catenin在四氯化碳诱导的小鼠肝纤维化过程中的定位表达及意义[J].解剖学报,2012,43(1): 97-102.

[25] Hohme S, Hengstler JG, Brulport M. Mathematical modelling of liver regeneration after intoxication with CCl(4). Chem Biol Int. 2007;168(1):74-93.

[26] Hora C, Romanque P, Dufour JF. Effect of sorafenib on murine liver regeneration. Hepatology. 2011;53(2):577-56.

[27] Safadi R,Friedman SL. Hepatic fibrosis-role of hepatic stellate cell activation. Med Gen Med. 2002;4(3):27-32.

[28] Kocabayoglu P, Friedman SL.Cellular basis of hepatic fibrosis and its role in inflammation and cancer. Front Biosci (Schol Ed). 2013;5:217-230.

[29] Berg T, DeLanghe S, Al Alam D, et al. β-catenin regulates mesenchymal progenitor cell differentiation during hepatogenesis. J Surg Res. 2010;164(2):276-285.

[30] Asahina K, Tsai SY, Li P, et al. Mesenchymal origin of hepatic stellate cells, submesothelial cells, and perivascular mesenchymal cells during mouse liver development. Hepatology. 2009;49(3):998-1011.

[31] Van Rossen E, Vander Borght S, van Grunsven LA, et al. Vinculin and cellular retinol-binding protein-1 are markers for quiescent and activated hepatic stellate cells in formalin-fixed paraffin embedded human liver. Histochem Cell Biol. 2009; 131(3):313-325.

[32] Zhao L, Burt AD. The diffuse stellate cell system.J Mol Histol. 2007;38(1):53-64.

[33] Kalinichenko VV, Bhattacharyya D, Zhou Y, et al. Foxfl+/-mice exhibit defective stellate cell activation and abnormal liver regeneration following CCl4 injury. Hepatology 2003;37: 107-117.

[34] Thomas RJ, Bhandari R, Barrett DA, et al. The effect of three dimensional co-culture of hepatocytes and hepatic stellate cells on key hepatocyte functions in vitro. Cells Tissues Organs. 2005;181:67-79.

[35] Gratzner HG. Monoclonal antibody to 5-bromo and 5-iododeoxyuridine:a new reagent for detection of DNA replication. Science. 1982;218:474-479.