人血管内皮生长因子-C基因治疗淋巴水肿的实验研究

Experimental study of gene therapy with human vascular endothelial growth factor-c in lymphedema

目的观察人血管内皮生长因子-C（VEGF-C）基因治疗阻塞性淋巴水肿的疗效。方法成年新西兰大白兔制成右后肢淋巴水肿模型;SD大鼠制成尾巴淋巴水肿模型。每种动物随机分2组,治疗组皮内注射人VEGF-C基因,对照组皮内注射生理盐水。测量治疗前后水肿部位体积变化;第3周取材,测VEGF-CmRNA及蛋白的表达水平;用淋巴管内皮细胞透明质酸受体-1（LYVE-1）抗体免疫组化染色观察淋巴管增生情况。结果基因治疗后兔右后肢体积明显减少,其减少值分别为：治疗组（24.40±1.08）ml,对照组（5.80±1.92）ml,差异有统计学意义（P=0.0001）;治疗组VEGF-CmRNA及蛋白明显表达,而对照组则不明显;治疗组淋巴管数量显著多于对照组（P=0.0004）,且管腔较粗。结论用VEGF-C基因治疗可诱导淋巴管生成,减轻或消除淋巴水肿。

Objective To study the efficacy of gene therapy with human vascular endothelial growth factor-c（VEGF-C） on obstructive lymphedema. Methods Two animal models of lymphedema were created： one in the right hind limb of adult New Zealand white rabbits and the other in SD mouse tail. Each model was randomly divided into two groups to receive intradermal injection of either VEGF-C gene（experimental group）, or saline（control group）. In rabbit model, the volume change of affected limb was measured. In mouse model, biopsy was performed after 3 weeks treatment to detect the expression of VEGF-C mRNA and proteins. The lymphangenesis was evaluated by immunohistochemical examination with lymphatic endothelium hyaluronan receptor antibody. Results The volume of the affect rabbit limb decreased by （24.40±1.08） ml in experimental group, compared with （5.80±1.92） ml in control group（P=0.0001）. The expression of VEGF-C mRNA and protein increased markedly in experiment group, but not in controls. More lymphatic vessels with large caliber were seen in experiment group （P=0.0004）.Conclusions VEGF-C gene therapy may alleviate or treat lymphedema by inducing lymangiogenesis.