脉血康对荷MHCC97H人肝癌裸鼠皮下移植瘤VEGF表达的影响

The Effect of Hirudin on the Expression of the Subcutaneous Implanted Tumor VEGF from Hepatocelluar Carcinoma MHCC97H in Athymic Mice

目的:研究脉血康(水蛭素)对荷MHCC97H人肝癌裸鼠皮下移植瘤VEGF表达的影响。方法:将60只接种高转移MHCC97H人肝癌裸鼠随机分成6组,每组10只。(1)阳性对照组:隔日腹腔注射2mg/kg顺铂注射液;(2)模型对照组:每日灌胃给予蒸馏水,0.2mL/10g体重;(3)低剂量脉血康组:脉血康每日口服灌胃0.5g/kg;(4)中剂量脉血康组:脉血康每日口服灌胃1.0g/kg;(5)高剂量脉血康组:脉血康每日口服灌胃2.0g/kg;(6)联合给药组:脉血康每日口服灌胃1.0g/kg,并隔日腹腔注射2mg/kg顺铂注射液。连续21天。用药后每3天各组裸鼠称重,并用游标卡尺测量每只裸鼠移植瘤的瘤结节最大直径a和最小径b,计算瘤体积,以相对肿瘤增殖率T/C%作为疗效评价指标。末次给药24h后处死动物,称重,拍照;分离瘤体,称重,甲醛固定。运用免疫组化技术检测瘤体VEGF的表达变化。结果与模型对照组比较,脉血康给药组、顺铂注射液组、联合给药组裸鼠皮下移植瘤的瘤重明显减少,有显著的统计学差异(P<0.001);联合用药组与不同剂量脉血康给药组及阳性对照组的裸鼠皮下移植瘤的瘤重比较有显著差异(P<0.05)。脉血康给药各组、联合给药组裸鼠皮下移植瘤的VEGF表达水平均低于模型对照组,差异有显著性意义(P<0.01);阳性对照组与模型对照组比较无统计学差异(P>0.05)。结论:脉血康对荷MHCC97H肝癌裸鼠皮下移植瘤的生长有明显的抑制作用,与顺铂联合应用作用更强。其作用机制与下调VEGF基因表达有关。

Objective ： To study the effect of hirndin on the expression of the subcutaneous implanted tumor VEGF from hepatocelluar carcinoma MHCC97H in athymic mice. Methods： Sixty athymic mice which bore high metastatic MHCC97 hepatocelluar carcinoma were randomized into six groups. Each group consists of ten mice. （ 1 ） Positive Control Group ： 2mg/kg cisplatin was injected into the mice＇ s abdominal cavity every alternate day; （2）Model Control Group：mice were lavaged daily with distilled water, O. 2mL/10g; （3）Low Dose Hirudin Group： mice were lavaged daily with hirudin 0. 5g/ kg; （4）Medium Dose Hirudin Group： mice were lavaged daily with hirudin 1.0g/kg; （5）High Dose Hirudin Group： mice were lavaged daily with hirudin 2.0g/kg; （6）Joint Group：mice were lavaged daily with hirudin 1.0g/kg and 2mg/ kg cisplatin was injected into their abdominal cavity every alternate day. The experiments lasted twenty - one days. After medication was applied, the athymic mice from each group were weighted every three days. In addition, the maximal diameter A and the minor diameter B of the tubercle of the implanted tumor in each athymic mouse were measured by verni- er caliper every three days. Therefore, the volume of the tumor and the relative rate of tumor reproduction （ T/C % ） were calculated and used as an indicator for the effectiveness of the treatment. Twenty - four hours after the last medication was applied, the tested animals were put down, weighted and photos were taken. Meanwhile, the tumor was separated from the body of each mouse. The separated tumor was weighed and was made rigid with methanol. The immunised technology was applied in order to measure the change of expression of the tumor VEGF. Results ： Compared with the Control Group, the weight of the subcutaneous implanted tumor in athymic mice of the Hirndin Group,the Cisplatin Injection group,theJoint Group had clearly reduced. The results show strong statistic differences （P 〈 0. 001 ）. The comparison of the weight of the subcutaneous implanted tumor in athymic mice among the Joint Group, various Hirudin groups and the Positive Control Group shows significant differences （P 〈 0. 05 ）. The VEGF expression level of the subcutaneous implanted tumor in athymic mice of various Hirudin groups and the Joint Group is lower than the Model Group. The results have significant meaning （P 〈0. 01 ）. There were no statistic differences between the comparison of the Positive Control Group and the Model Group （P 〉 0.05 ）. Conclusion： The hirudin has evidently inhibited the growth of the subcutaneous implanted tumor from hepatocelluar carcinoma MHCC97 in athymic mice. The application of the combination of hirudin and cisplatin is more effective. The mechanism is relevant with the expression of down - regulating genes VEGF.