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目的通过检测血管内皮生长因子(VEGF)在大鼠肝再生过程中表达量的变化,探讨VEGF在肝再生中的作用。方法300只SD大鼠随机分为正常对照组、假手术组和实验组。实验组大鼠切除2/3肝,分别于术后不同时段取肝组织或原位灌注分离肝实质细胞。用免疫组织化学和Western blotting技术检测肝组织和肝实质细胞中VEGF的表达变化。结果正常肝组织的VEGF阳性细胞很少;肝切除(PH)后24 h阳性细胞数显著增加(P<0.01),主要分布于门管区周围;PH 72 h时阳性细胞数达到高峰,几乎遍及整个肝小叶;从PH 120h起VEGF阳性细胞数逐渐减少至正常。Western blotting检测结果表明,肝实质细胞的VEGF表达量于PH 0~12 h较少,PH 12 h后增多,PH 24~72 h的表达量约为PH 0~12 h的2倍,PH72 h后逐渐下降,至120 h恢复正常;肝组织VEGF的表达量于PH 0~12 h较少,PH 12 h后逐渐增多,PH 72 h时出现高峰,约为PH 0~12 h的4倍;肝组织VEGF的表达量大于肝实质细胞的表达量。结论VEGF可能是参与肝脏组织结构重建的重要的生长因子:肝实质细胞是肝中VEGF的重要来源之一。
Objective To explore the role of VEGF in liver regeneration by detecting the expression of vascular endothelial growth factor (VEGF) in rat liver regeneration. Methods 300 SD rats were randomly divided into normal control group, sham operation group and experimental group. Experimental group rats were excised 2/3 liver, liver tissue was taken at different time points after operation, or perfused to isolate hepatic parenchymal cells. Immunohistochemistry and Western blotting were used to detect the expression of VEGF in liver tissue and liver parenchyma cells. Results There were only a few VEGF positive cells in normal liver tissue. The numbers of positive cells at 24 h after hepatectomy (PH) were significantly increased (P <0.01), mainly distributed around portal area. The number of positive cells reached the peak at 72 h after PH, Hepatic lobules; the number of VEGF-positive cells gradually decreased to normal from PH 120h. The results of Western blotting showed that the expression of VEGF in hepatic parenchymal cells was less in PH0-12h, increased in PH12h, and the expression level in hepatocytes was about twice as high as PH0-12h after PH24-72h Gradually decreased and returned to normal after 120 h. The expression of VEGF in liver tissue was less in PH 0-12 h, increased gradually after 12 h PH, and peaked at 72 h, which was about 4 times of PH 0-12 h. Tissue VEGF expression is greater than the expression of hepatic parenchymal cells. Conclusion VEGF may be an important growth factor involved in the structural remodeling of liver tissue. Hepatic parenchymal cells are one of the important sources of VEGF in the liver.