| Objective: To investigate the effects of exogenous VEGF on vascular regeneration and reconstruction of the splenic tissues during the process of splenic autotransplantation into the greater omentum, and explore the feasibility and mechanisms of exogenous VEGF in vascular regeneration and reconstruction of the splenic tissues in splenic autotransplantation into the greater omentum, and offer theoretical reference rationale of exogenous VEGF for experimental study and clinical application of splenic autotransplantation .Methods: Healthy Kunming mice were randomized into the experimental groups with splenectomy and splenic autotransplantation and the control groups with sham-operation, then the experimental groups with splenectomy and splenic autotransplantation were randomly divided into two groups: (1) the experimental observation groups with exogenous VEGF;(2) the control observation groups without exogenous VEGF. The mice in experimental observation groups were injected with exogenous VEGF into there caudal veins on day 7,15,30,60 respectively after operation and three mice of them were killed and their splenic tissus were taken as specimen on day 7,14,30,60,90 after injected with exogenous VEGF; On the correspondent day above, three mice were killed and their splenic tissus were taken as specimen in the control observation groups without exogenous VEGF and the control groups with sham-operation respectively . The following investigations were conducted in each group: (1) Observing the morphologic changes in autotransplanted splenic tissues: specimen slice made according to routine were observed under light microscope and electron microscope.(2) Testing the area density changes of renascent blood vessels in splenic tissues: after intubatton was conducted through left ventricle and right atria was opened and lavaged ,the splenic tissues slice made by 30 minutes' the perfusion of Chinese ink into aorta under the same pressure was analyzed and the area density was tested by using computer image system . (3) KDR test: the density changes of KDR positive cells were tested by combining immunohistochemical(IHC) methods and computer image analysis qualification.(4) The test of KDR mRNA: expressional changes of KDR mRNA were investigated and analyzed by the in situ hybrdization (ISH) methods to study exogenous VEGF on expressional changes of KDR mRNA.Results: 1,The morphological observation: (1) on day 7,14,30 after injection with exogenous VEGF, the mice in the experimental observation groups were found much more renascent blood vessels,splenic corpuscles,central arteries and peripheral lymphatic sheathes in the autotransplanted splenic tissues than those found in the control observation groups without exogenous VEGF , and eventually formed more integra structure of white pulp,marginal zone and red pulp with much less fibrous tissue. (2) There was no significantly difference in histological characters between groups with exogenous VEGF on day 60 after injection and the control observation groups without exogenous VEGF. 2,Testing the area density changes of renascent blood vessels in splenic tissues: (1) On day 7,15,30 after injection the ink showed that renascent blood vessels are much more in number higher in area density test in splenic tissues on the VEGF group than those on the control group. (2) On day 60 after operation it showes no significant difference in the number of renascent blood vessels between the VEGF group and the control group. 3,The test of KDR: (1)On day 7,15,30, the area density of the VEGF group was higher than the control group. (2) On day 60 after operation, there was no significant difference between the two groups in the KDR area density. 4,KDR mRNA test: (1)Expressional changes of KDR mRNA investigated in autotransplantated splenic tissues on day 7,14,30 of the experimental observation groups with exogenous VEGF showed greater than those in the control observation groups without exogenous VEGF. (2)Expressional changes of KDR mRNA investigated in autotransplantated...
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