Vascular endothelial growth factor and angiopoietins regulate sinusoidal regeneration and remodeling after partial hepatectomy in rats
Vascular endothelial growth factor and angiopoietins regulate sinusoidal regeneration and remodeling after partial hepatectomy in rats
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DOI：
10.3748/wjg.v11.i46.7254
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发表时间：
2005-12-14
影响因子：
4.3
通讯作者：
Miyazaki, Masaru
中科院分区：
文献类型：
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作者：
Shimizu, Hiroaki;Mitsuhashi, Noboru;Miyazaki, Masaru
目的：为了研究肝部分切除术后肝窦再生的调控机制，方法：我们研究了血管生成素（Ang）-1，Ang-2，Tie-2和血管内皮生长因子（VEGF）在再生肝组织中的表达，通过定量逆转录聚合酶链反应（RT-PCR）使用LightCycler（Roche Diagnostics）和免疫组化染色后70%肝切除大鼠。下一步，我们通过原位胶原酶灌注和逆流淘洗从再生肝组织中分离肝细胞（肝细胞、肝窦内皮细胞（SEC）、枯否细胞和肝星状细胞（HSC）），以确定肝切除术后这些血管生成因子的潜在细胞来源。增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）染色和末端脱氧核苷酸转移酶缺口末端标记（terminal deoxynucleotidyl transferase d-uridine triphosphate nick end labeling，TUNEL）法检测SECs的增殖和凋亡。Ang-1 mRNA在肝切除前即有表达，但表达缓慢增加，96 h达高峰。Ang-2 mRNA在肝切除前几乎不表达，而在120和144 h时表达明显增加。在分离的细胞中，VEGF mRNA的表达被发现主要在肝细胞部分。同时，Ang-1和Ang-2的mRNA在SEC和HSC组分中被发现，但在后者中更突出。SECs的PCNA标记指数缓慢增加，72 h达高峰，而凋亡的SECs则在120 ~ 144 h出现。结论：Ang-Tie系统与VEGF共同参与调节肝切除术后肝窦再生过程中SEC增殖与凋亡的平衡。然而，VEGF系统在窦状隙再生的早期阶段比Angiopoietin/Tie系统起更重要的作用。(C)2005年WJG出版社和Elsevier Inc. All rights reserved.
AIM: To study the regulatory mechanisms of sinusoidal regeneration after partial hepatectomy.METHODS: We investigated the expression of angiopoietin (Ang)-1, Ang-2, Tie-2, and vascular endothelial growth factor (VEGF) in regenerating liver tissue by quantitative reverse-transcription polymerase chain reaction (RT-PCR) using a LightCycler (Roche Diagnostics) and also immunohistochemical staining after 70% hepatectomy in rats. In the next step, we isolated liver cells (hepatocytes, sinusoidal endothelial cell (SEC), Kupffer cell, and hepatic stellate cells (HSC)) from regenerating liver tissue by in situ collagenase perfusion and counterflow elutriation, to determine potential cellular sources of these angiogenic factors after hepatectomy. Proliferation and apoptosis of SECs were also evaluated by proliferating cell nuclear antigen (PCNA) staining and the terminal deoxynucleotidyl transferase d-uridine triphosphate nick end labeling (TUNEL) assay, respectively.RESULTS: VEGF mRNA expression increased with a peak at 72 h after hepatectomy, decreasing thereafter. The expression of Ang-1 mRNA was present at detectable levels before hepatectomy and increased slowly with a peak at 96 h. Meanwhile, Ang-2 mRNA was hardly detected before hepatectomy, but was remarkably induced at 120 and 144 h. In isolated cells, VEGF mRNA expression was found mainly in the hepatocyte fraction. Meanwhile, mRNA for Ang-1 and Ang-2 was found in the SEC and HSC fractions, but was more prominent in the latter. The PCNA labeling index of SECs increased slowly, reaching a peak at 72 h, whereas apoptotic SECs were detected between 120 h and 144 h.CONCLUSION: Ang-Tie system, together with VEGF, plays a critical role in regulating balance between SEC proliferation and apoptosis during sinusoidal regeneration after hepatectomy. However, the VEGF system plays a more important role in the early phase of sinusoidal regeneration than angiopoietin/Tie system. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.