探讨活化态肝星状细胞(aHSC)调控肝细胞癌（肝癌）微血管形成的机制。

组织标本来源于2013年1月至12月在中山大学附属第三医院肝胆外科行肝切除术的25例肝癌患者。所有患者均签署知情同意书，符合医学伦理学规定。其中男21例，女4例；年龄29～75岁，中位年龄54岁。分别取肝癌组织（肝癌组）、癌旁组织（癌旁组），同时选取21例正常肝组织作对照组。免疫组织化学方法（免疫组化法）检测平滑肌肌动蛋白（SMA）、血管生成素（Ang） -1、分化群（CD）34表达。蛋白质印迹法（Western blot）检测Ang-1表达水平。肝癌组织SMA高表达为aHSC的标志。3组比较采用单因素方差分析和LSD-t检验。蛋白表达的相关性采用Spearman相关分析。

肝癌组、癌旁组和对照组SMA平均光密度值分别为（4.56±0.64）×10⁴、（2.71±0.37）×10⁴、（2.25±0.48）×10⁴，肝癌组明显高于癌旁组和对照组（LSD-t=7.09，7.42；P<0.05）。3组Ang-1光密度值分别为（3.11±0.27）×10⁵、（2.28±0.20）×10⁵、（1.26±0.15）×10⁵，肝癌组明显高于癌旁组和对照组（LSD-t=3.00，3.14；P<0.05）。3组Ang-1表达相对灰度值分别4.33±1.17、1.62±0.33、1.60±0.38，肝癌组明显高于癌旁组和对照组（LSD-t=2.71，2.74；P<0.05）。3组CD34光密度值分别为（18.30±0.36）×10³、（5.75±1.17）×10³、（2.75±0.72）×10³，肝癌组明显高于癌旁组和对照组（LSD-t=3.21，3.36；P<0.05）。SMA与Ang-1、CD34，Ang-1与CD34两两之间均呈正相关（r= 0.442，0.449，0.582；P<0.05）。

肝癌组织中SMA、Ang-1、CD34呈高表达，且三者密切相关。肝癌组织中存在SMA标记的aHSC，aHSC可能通过分泌Ang-1促进肝癌微血管形成。

To investigate the regulatory mechanism of activated hepatic stellate cell (aHSC) in the microvascular generation of hepatocellular carcinoma (HCC).

Tissue samples of 25 patients with HCC who underwent liver resection in Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University from January to December 2013 were collected. The informed consents of all patients were obtained and the ethical committee approval was received. There were 21 males and 4 females with age ranging from 29 to 75 years old and the median age of 54 years old. The HCC tissues (HCC group) and peri-cancer tissues (peri-cancer group) were collected and normal liver tissues of 21 cases were collected as control group. The expressions of smooth muscle actin (SMA), angiogenin (Ang-1), and cluster of differentiation (CD) 34 were detected by immunohistochemistry. The expression level of Ang-1 was detected by Western blot. The high expression of SMA in HCC tissues was regarded as a marker of aHSC. The differences of 3 groups were compared using one-way analysis of variance and LSD-t test. Spearman correlation analysis was used to analyze the correlations between different proteins expression.

The mean of optical densities of SMA in HCC, peri-cancer, control group were (4.56±0.64)×10⁴, (2.71±0.37)×10⁴, (2.25±0.48)×10⁴ respectively, which was significantly higher in HCC group than those in peri-cancer group and control group (LSD-t=7.09, 7.42; P<0.05). The optical densities of Ang-1 in 3 groups were (3.11±0.27)×10⁵, (2.28±0.20)×10⁵, (1.26±0.15)×10⁵ respectively, which was significantly higher in HCC group than those in peri-cancer group and control group (LSD-t=3.00, 3.14; P<0.05). The relative gray values of Ang-1 expression in 3 groups were 4.33±1.17, 1.62±0.33, 1.60±0.38 respectively, which was significantly higher in HCC group than those in peri-cancer group and control group (LSD-t=2.71, 2.74; P<0.05). The optical intensities of CD34 in 3 groups were (18.3±0.36)×10³, (5.75±1.17)×10³, (2.75±0.72)×10³ respectively, which was significantly higher in HCC group than those in peri-cancer group and control group (LSD-t=3.21, 3.36; P<0.05). There were positive correlations between SMA, Ang-1, CD34 (r=0.442, 0.449, 0.582; P<0.05).

High expressions of SMA, Ang-1 and CD34 can be observed in HCC group and closely related with each other. SMA marked aHSC can be observed in the HCC tissues. The aHSC may promote the microvascular generation of HCC by secreting Ang-1.