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作 者:彭瑞云[1] 高亚兵[1] 熊呈琦[1] 秦全红[1] 王德文[1]
机构地区:[1]军事医学科学院放射医学研究所八室,北京100850
出 处:《细胞与分子免疫学杂志》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
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.
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