Vasuri F, Fittipaldi S, Abualhin M, Degiovanni A, Gargiulo M, Stella A, Pasquinelli G
Pathology Unit, Department of Experimental, Diagnostic and Specialty Medicine, S. Orsola-Malpighi Hospital, Bologna University, Italy; Vascular Surgery Unit, Department of Experimental, Diagnostic and Specialty Medicine, S. Orsola-Malpighi Hospital, Bologna University, Italy.
Vascular Surgery Unit, Department of Experimental, Diagnostic and Specialty Medicine, S. Orsola-Malpighi Hospital, Bologna University, Italy.
Eur J Vasc Endovasc Surg. 2014 Oct;48(4):430-7. doi: 10.1016/j.ejvs.2014.05.002. Epub 2014 Jun 16.
Hepatocyte growth factor (HGF), the c-Met receptor, and hypoxia-inducible factor (HIF) are crucial for regenerative processes including ischemic wound healing. The aims of the present study are (a) to analyze the tissue c-Met and HIF-1α expression in skin from patients with critical limb ischemia (CLI); (b) to compare the serum HGF levels of CLI and control subjects.
This is a prospective, controlled, single-center study. Thirty-seven patients were enrolled. A skin sample adjacent to the ischemic lesion was taken from 20 patients with CLI; skin samples were taken from the surgical wounds of 17 patients surgically treated for abdominal aortic aneurysm as healthy controls. Serum samples were taken in all cases. Samples were formalin fixed, paraffin embedded, and routinely processed. Tissue inflammation was histologically assessed. Immunohistochemistry was performed with antibodies against total c-Met receptor, activated Met (p-Met), and HIF-1α. RT-polymerase chain reaction was used to quantify HIF-1α mRNA. The enzyme-linked immunosorbent assay was performed to evaluate serum HGF levels.
With immunohistochemistry, while total c-Met was unchanged, different patterns of p-Met positivity were observed between CLI and control cases (p < .001). In particular, CLI skin showed a total negativity or membrane positivity for p-Met (19/20 cases), while control skin mainly showed cytoplasmic positivity in the epidermal basal layer (16/17 cases). HIF-1α was diffusely lost in CLI, but HIF-1α mRNA was threefold higher than in controls. Finally, mean serum HGF levels were 590.5 pg/mL and 2380.0 pg/mL in CLI and control groups respectively (p < .001).
In CLI patients a significant decrease in serum HGF levels, concomitant with a loss of skin HIF-1α stabilization and a lack of c-Met phosphorylation were seen, probably driving a decrease in wound-healing functions. The next hypothesis is that HGF application might reactivate the c-Met receptor, stabilizing the normal wound healing process.
肝细胞生长因子（HGF）、c-Met受体和缺氧诱导因子（HIF）对于包括缺血性伤口愈合在内的再生过程至关重要。本研究的目的是：（a）分析严重肢体缺血（CLI）患者皮肤组织中c-Met和HIF-1α的表达；（b）比较CLI患者和对照受试者的血清HGF水平。
这是一项前瞻性、对照、单中心研究。共纳入37例患者。从20例CLI患者缺血病变附近采集皮肤样本；从17例接受腹主动脉瘤手术治疗的患者手术伤口采集皮肤样本作为健康对照。所有病例均采集血清样本。样本用福尔马林固定、石蜡包埋并常规处理。对组织炎症进行组织学评估。用抗总c-Met受体、活化Met（p-Met）和HIF-1α的抗体进行免疫组织化学检测。采用逆转录聚合酶链反应定量HIF-1α mRNA。采用酶联免疫吸附试验评估血清HGF水平。
免疫组织化学检测显示，虽然总c-Met无变化，但CLI患者和对照病例之间观察到不同的p-Met阳性模式（p<0.001）。具体而言，CLI患者皮肤p-Met呈完全阴性或膜阳性（19/20例），而对照皮肤主要在表皮基底层呈细胞质阳性（16/17例）。HIF-1α在CLI患者中弥漫性缺失，但HIF-1α mRNA比对照高3倍。最后，CLI组和对照组的平均血清HGF水平分别为59
