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目的研究ＶＥＧＦ 基因在正常和放射复合伤口愈合中的表达及意义，探讨其在血管再生中的作用。方法将４ ８ 只Ｗｉｓｔａ ｒ 二级大鼠背部皮肤造成圆形伤口后，以２５Ｇｙ ６０ Ｃｏ γ射线局部照射，于伤后２ 、５ 、１０ 、１５ 、２ １ 和２８ｄ 活杀取材，采用免疫组织化学、原位杂交和原位ＰＣ Ｒ 等方法，研究ＶＥＧＦ 基因的表达及意义。结果单纯创伤组于伤后２ｄ，新生血管内皮细胞浆内ＶＥＧＦ 的表达呈强阳性；５ ｄ 时毛细血管数明显增多，其内皮细胞胞浆内ＶＥＧＦ 表达呈阳性；而于１０ｄ后，ＶＥＧＦ 的表达逐渐减少。创伤＋照射组则于伤后５ｄ，ＶＥＧＦ 的表达在血管内皮细胞胞浆内呈弱阳性，１０ ｄ 时阳性，１５ ｄ 后则呈弱阳性。原位杂交显示，ＶＥＧＦｍＲＮＡ 于单纯创伤组伤后２ ～５ ｄ 及创伤＋照射组伤后５ ～１０ｄ，血管内皮细胞浆内阳性。原位反转录ＰＣ Ｒ 则显示，ＶＥＧＦｍＲＮ Ａ 于单纯创伤组伤后２ ～１５ｄ 及创伤＋照射组伤后５ ～２８ｄ，呈阳性反应。结论内源性ＶＥＧＦ 基因的表达参与创伤愈合中血管再生过程。辐射使血管内皮细胞中ＶＥＧＦ 的表达减少。原位反转录ＰＣ Ｒ 方法较原位杂交能客观地反映内源性ＶＥＧＦ 基因的表达情况。
Objective To study the expression and significance of VEGF gene in normal and radiation wound healing and to explore the role of VEGF gene in revascularization. Methods Forty-eight Wistar rats were divided into two groups. The wounds on the back of the rats were made into circular wounds. The wounds were irradiated with 25Gy 60 Co γ ray and killed at 2, 5, 10, 15, 21 and 28 days after injury. Chemistry, in situ hybridization and in situ PC R methods to study the expression of VEGF gene and its significance. Results The expression of VEGF was significantly positive in neovascular endothelial cells at 2 days after injury in pure trauma group. The number of capillary vessels was significantly increased at 5 days, and the expression of VEGF was positive in the cytoplasm of endothelial cells. At 10 days, the expression of VEGF gradually decreases. At 5 days after wounding, the expression of VEGF was weakly positive in the cytoplasm of vascular endothelial cells and positive in 10 days after injury and weakly positive after 15 days. In situ hybridization showed that VEGF mRNA was positive in the plasma of endothelial cells 2 to 5 days after wound healing and 5 to 10 days after wounding + irradiation. In situ reverse transcription of PC R showed that VEGFmRN A was positive in 2 ~ 15 days after trauma group and 5 ~ 28 days after trauma + irradiation group. Conclusion The expression of endogenous VEGF gene is involved in the process of blood vessel regeneration during wound healing. Radiation reduces the expression of VEGF in vascular endothelial cells. In situ reverse transcription PC R method compared with in situ hybridization can objectively reflect the expression of endogenous VEGF gene.