Analysis of the role of angiogenesis and non-parenchymal cells during hepatic regeneration.

我们用HGF拮抗剂（NK4）和VEGF受体抑制剂（sFlt-1）研究了抗血管生成对肝再生的影响。表达NK4的腺病毒载体（Ad.NK4）或表达sFlt-1的腺病毒载体（Ad.sFlt-1）分别感染小鼠肝脏。70%的患者在感染后48小时进行肝切除术。以表达LacZ的腺病毒载体（Ad.LacZ）作为对照。Nk4或sFlt-1的表达在感染后48小时达到最高水平（Nk4: 300ng/ml, sflt:350ng/ml），并持续2周。在部分肝切除术后12、24、36、48、72和120小时切除残肝，评估肝再生情况。结果表明，Ad组肝残重的增加明显延迟。NK4或Ad。sFlt-1感染小鼠。有丝分裂细胞和BrdU指数显示Ad肝再生明显延迟。NK4或Ad。肝切除术后48或36小时sFlt-1感染小鼠。CD31免疫组化染色显示Ad窦内皮细胞生长受到抑制。NK4感染小鼠。Ang-1、Ang-2、Tie-1和Tie-2被认为在血管成熟过程中起着至关重要的作用。Real time PCR结果显示，Ang-1、Ang-2、Tie-1、Tie-2 mRNA的表达受到抑制，差异有统计学意义。提示肝切除术后肝再生与血管生成密切相关。

We investigated the influence of anti-angiogenesis for hepatic regeneration using HGF antagonist (NK4) and suppressor of VEGF receptor (sFlt-1). NK4 expressing adenoviral vector (Ad.NK4) or sFlt-1 expressing adenoviral vector (Ad.sFlt-1) were infected mice liver. Seventy percent hepatectomy was performed 48 hours after infection. LacZ expressing adenoviral vector (Ad.LacZ) was used as control. Expression of Nk4 or sFlt-1 reached maximum level 48 hours after infection (NK4: 300ng/ml, sFlt-1:350ng/ml) and sustained for 2 weeks. Remnant liver was removed at 12, 24, 36, 48, 72 and 120 hrs after partial hepatectomy, and hepatic regeneration was assessed. As a result, increase of remnant hepatic weight was significantly delayed in Ad.NK4 or Ad.sFlt-1 infected mice. Results of mitotic cells and BrdU index showed significant delay of hepatic regeneration in Ad.NK4 or Ad.sFlt-1 infected mice at 48 or 36 hours after hepatectomy. Immnuhistochemical staining of CD31 showed suppression of sinusoidal endothelial cells growth in Ad.NK4 infected mice. Ang-1, Ang-2, Tie-1, and Tie-2 are considered to be crucial in the process of maturation of vessels. Real time PCR showed suppression of mRNA of Ang-1, Ang-2, Tie-1, and Tie-2 with significant difference. These results suggested that hepatic regeneration after hepatectomy closely correlates with angiogenesis.