外源性VEGF对自体移植脾组织血管再生的影响
VEGF,,],自体移植脾组织；血管再生；VEGF；KDR；小鼠,1材料和方法,2结果,3讨论,[参考文献]
[摘 要] 目的 探讨应用外源性VEGF促进自体脾组织移植后血管再生与结构重建的影响，为VEGF用于自体移植脾组织的实验研究和临床应用提供理论参考依据。方法 采用216只昆明种小白鼠随机分为脾切除自体脾组织移植组和假手术组，再将脾移植组分为实验组和对照组。实验组小鼠分别于术后7、14、30、60 d于尾静脉内注射VEGF，注射后7、15、30、60、90 d取脾组织标本。进行组织学观察、血管面密度测定、免疫组化染色方法KDR的检测、KDR阳性细胞密度测定、组织原位杂交技术KDR mRNA检测。结果术后7、15、30 d注射VEGF组 注射VEGF后较对照组的血管数量明显增多；术后60 d注射VEGF组注射VEGF后与对照组各时相点的组织学特征没有明显差别；术后7、15、30 d注射VEGF组KDR面积密度值高于对照组，术后60 d注射VEGF组 KDR面密度值与对照组比较无显著差别；术后7、15、30 d注射 KDR mRNA表达阳性细胞密度高于对照组，术后60 d注射VEGF组 KDR mRNA表达阳性细胞密度与对照组比较没有明显差异。结论 外源性VEGF能够促进移植脾组织内血管生长，自体脾组织移植术后7~15 d开始静脉应用VEGF是促进血管再生的较理想时间；外源性VEGF可能是通过与移植脾组织内所表达的KDR结合发挥作用。[关键词] 自体移植脾组织；血管再生；VEGF；KDR；小鼠
Effects of exogenous VEGF on vascular regeneration in autotransplanted splenic tissues of mice
GUO Guangjin, JIANG Dengjin, JIANG Heng, ZUO Yanfang
(Department of Surgical Anatomy and Surgery, College of Medicine, Third Military Medical University, Chongqing 400038, China )
Abstract: Objective To investigate the effects, the feasibility and the mechanism of exogenous VEGF on vascular regeneration and reconstruction of the splenic tissues during the process of splenic autotransplantation into the greater omentum. Methods A total of 216 healthy Kunming mice were randomized into splenic tissue autotransplantation+VEGF group (named as VEGF group), splenic tissue autotransplantation group (control group) and sham operation group. The mice in VEGF group were injected with exogenous VEGF into the caudal veins on day 7, 15, 30, 60 respectively after operation. Three mice of 3 groups were killed and the splenic tissue was taken as specimen on day 7, 14, 30, 60, 90 after injection of exogenous VEGF to conduct the following investigations. The morphologic changes in autotransplanted splenic tissue were observed under light microscope and electron microscope. After intubation was conducted through left ventricle and right atria was opened and lavaged with Chinese ink, the splenic tissue pieces were made and the area density of blood vessel was tested by using computer image system. The density of KDR positive cells was tested by immunohistochemical methods and computer image analysis. The expression of KDR mRNA were investigated and analyzed by in situ hybridization to study exogenous VEGF on KDR mRNA expression. Results On day 7, 15, 30 after injection with exogenous VEGF, the mice in VEGF group were found more renascent blood vessels, splenic corpuscles, central arteries and peripheral lymphatic sheathes in the autotransplanted splenic tissues than the control group, and eventually formed more integral structure of white pulp, marginal zone and red pulp with much less fibrous tissue, while on day 60 there was no significant difference between VEGF group and the control group. On day 7, 15, 30 after injection, the renascent blood vessels were more in number, the KDR area density and the KDR mRNA expression in splenic tissues were higher in VEGF group than the control group, while on day 60 there were no significant differences in the area density of renascent blood vessel, the KDR area density and the KDR mRNA expression between the two groups. Conclusion The exogenous VEGF can enhance the vascular regeneration and reconstruction of the autotransplanted splenic tissues into the greater omentum. The day 7 to day 15 after the autotransplantation of splenic tissues was the ideal period for the venous injection of exogenous VEGF that can make its effects through elevating the expression of KDR mRNA. ......
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