摘要
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。
Aim To observe the expression of VEGF in n ormal and radi-ation skin wound healing and investigate the significance in the capilla ry regeneration.Methods Full thickness dorsum skin of 48Wistar rats were excised at diameter 1.5cm to form a wound.After wound,these rats in radiation group were irradia tedly local immediately with a dose o f25Gy 60Co γrays.The specimens were obtained 2,5,10,15,21and 28days after injury and the expre ssion of VEGF was investigatedby means of immunohistochemical sta ining,in situ hybridization(ISH)and in situ RT-PCR(IS RT-PCR).Results In normal group,the ex-pression of VEGF protein was strongly positive in the plasma of en-dothelial cells of new-formed capillaries on 2days,and VEGF still pos-itive in these cells on 5-10days and p ositive degree decreased 15days after wound;Whereas in radiation pl us wound group,The expression of VEGF protein was slightly positive i n the endothelial cells on 5days,positive on 10days and slightly positive after 15days.ISH demonstrat-ed that the expression of VEGF mRNA wa s positive in the plasma of endothelial cells on 2-5days in norm al wound group and on 5-10days in radiation plus wound group,where as IS RT-PCR demonstrated that the expression of VEGF mRNA was positive in the endothelial cells on2-15days in normal wound group and on5-28days in radiation plus wound group.Conclusion The endogenous VEGF gene expression play an important role during capillary r egeneration.Radiation may de-creased the expression of VEGF gene.The IS RT-PCR may more objec-tively reflect the expression of VEG F.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第5期410-412,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
全军"九五"重点招标课题基金资助
No.96Z008
