自体移植脾组织VEGF、KDR表达与血管再生的实验研究

目的 研究自体移植脾组织血管再生及VEGF、KDR表达规律，阐明VEGF、KDR对移植脾组织血管再生的调控作用，为脾脏外科临床及实验研究提供理论依据。方法 健康Wistar大鼠70只，体重100—120g，随机分为7组，每组10只中又设脾切除自体脾移植组5只，假手术组5只，分别于术后7，14，30，60，90，120，180d进行：(1)自体移植脾组织病理学检测；(2)大鼠行主动脉插管灌注墨汁，光镜观测再生血管并采用图像分析测定其密度；(3)免疫组化抗VEGF、KDR抗体染色，图像分析定量，阐明其表达规律及与血管再生的关系。结果 (1)自体脾组织移植术后7d即有血管从大网膜向脾组织内伸展，移植脾组织内血管密度逐渐增大，至术后180d血管再生接近正常；(2)自体脾组织移植术后7d、14d，VEGF、KDR阳性染色细胞密度迅速升高，术后60d达高峰，以后逐渐降低，至术后180d VEGF、KDR阳性染色细胞密度趋向正常。结论 自体脾组织大网膜内移植术是简便有效的脾移植方法；移植脾组织新生血管由大网膜再生而来；术后移植脾组织内VEGF、KDR表达量升高，促进血管形成，血管再生完成后恢复正常水平。

An experimental study on regeneration of blood vessels and expression of VEGF and KDR in splenic auto-transplantation

Objective To study the regeneration process of blood vessels in splenic autotransplants and analyze the regulatory effects of VEGF and KDR on vascular regeneration in autotransplanted spleen. Methods Seventy healthy Wistar rats were equally divided into 7 groups. In each group, splenectomy and splenic autotransplantation were performed in 5 rats and the left 5 received only sham operation. The following determinations were conducted 7, 14, 30, 60, 90, 120 and 180 days later in each group: (1) Pathological changes of splenic autotransplants with light microscopy and transmission electron microscopy;(2) Density changes of renascent blood vessels after splenic autotransplantation with light microscopy and computerized imaged analysis qualification 24 h after the perfusion of Chinese ink into the aorta;(3) VEGF and KDR positive cells with immunohistochemistry. Meanwhile, computerized image analysis was used to investigate their density changes. Results (1) Seven days after the operation, renascent blood vessels were seen to extend from greater omentum into splenic autografts. The vascular density increased with time prolongation. One hundred and eighty days after the operation, the density almost became nornmal. (2) Density of VEGF and KDR positive cells rose quickly 7 days after the operation. The peak density was noticed 60 days after the operation and it gradually decreased to normal after the time phase. Conclusions Autotransplantation of spleen into greater omentum is a convenient and effective way for splenic transplantation. After autotransplantation, the blood vessels complete their regeneration. The renascent blood vessels come from greater omentum and extend into splenic autografts. In early stage after the operation, the expression of VEGF and KDR in autotransplanted spleen quickly increase and instently prompts the vascular regeneration. When the regeneration process of blood vessels finishes, the expression of VEGF and KDR in the splenic autotransplants decreases to normal.