应用肿瘤双植入模型技术研究扶正消癥方的抑瘤作用及对IL-12,MMP-9表达的影响

Effects of Fuzheng Xiaozheng Formula on Expression of IL-12 and MMP-9 in Mice with Transplanted Liver Cancer Investigated by Double-grafted Tumor System

目的:研究扶正消癥方(含药血清)对肝癌H22荷瘤小鼠的抑制肿瘤血管生成及抗肿瘤转移的作用。方法:昆明种小鼠制备正常小鼠含药血清、正常小鼠对照血清、荷瘤小鼠含药血清、荷瘤小鼠对照血清,应用肿瘤双植入模型技术建立小鼠H22肝癌移植瘤模型,除空白组外,把已制备好的各组血清分别于第4,5,6天向右侧肿瘤内注射,观察双侧肿瘤的变化并分别计算出抑瘤率,用酶联免疫吸附(ELISA)法测定血清中白细胞介素-12(IL-12)的含量,用免疫组化法结合图像分析技术检测肿瘤组织中基质蛋白金属酶-9(MMP-9)的含量。结果:扶正消癥方含药血清可使荷瘤小鼠双侧肿瘤明显缩小,与对照血清组比较(P<0.01);正常动物血清与模型动物病理血清对肿瘤作用的差异无统计学意义。扶正消癥方可提高IL-12的水平并降低肿瘤组织中MMP-9的表达,与对照血清组比较(P<0.01)。结论:扶正消癥方具有抑制肿瘤血管生成和抗肿瘤转移的作用,调节IL-12的水平并抑制MMP-9的表达可能是其机制之一。

Objective： To study the effects of Fuzheng Xiaozheng Formula（pharmaceutic serum） in mice with transplanted liver cancer（H22） on inhibiting angiogenesis and metastasis.Method： mice were randomly divided into 4 groups,to prepare 4 kinds of serums,including pharmaceutic serum of normal mice,control serum of normal mice,pharmaceutic serum of tumor-bearing mice,control serum of tumor-bearing mice.Model with transplanted liver cancer（H22） in mice was made by double-grafted tumor system.We injected the serum to each group on the 4,5,6 day except the normal group,and observed the tumors changes of each side and calculated the inhibitory of tumor（IR）.Concentration of IL-12 was determined by enzyme linked immunosorbent assay（ELISA）,immune histochemistry and graphical analysis were used to detect the expression of matrix metalloproteinase-9（MMP-9）of tumor tissues.Result： The serum of Fuzheng Xiaozheng Formula had positive effects in inhibiting the growth of the double flank tumor,it showed differences from the control serum group（P 0.01）.The serum of tumor-bearing mice as no differences from the serum of normal mice.The group of Fuzheng Xiaozheng Formula had positive effects in raising the expression of IL-12 and inhibiting the expression of MMP-9 in tissue.It showed differences from the control serum group（P 0.01）.Conclusion： One of mechanisms responsible for Fuzheng Xiaozheng Formula＇s effect on angiogenesis and metastasis of tumor might be related to regulating the expression of IL-12 and inhibiting the expression of MMP-9.