人参皂苷Rg3对小鼠大肠癌细胞的抗肿瘤活性研究

Anticancer Effect of Ginsenoside Rg3 on Mice Colorectal Cancer

目的从体外实验到体内实验2个层面观察人参皂苷Rg3对大肠癌细胞的抗肿瘤效果。方法体外培养小鼠大肠癌细胞(CT26细胞),采用四甲基偶氮唑蓝比色法分析人参皂苷Rg3对CT26细胞生长的影响。皮下接种建立BALB/c小鼠大肠癌模型。待肿瘤可扪及(直径约3 mm)时,将荷瘤鼠随机分为生理盐水组及人参皂苷Rg3浓度5、10、20 mg/kg组。每周3次测量小鼠肿瘤长、宽及小鼠体质量,绘制肿瘤体积增长曲线和体质量变化曲线;记录小鼠生存时间,绘制生存曲线。观测小鼠精神状况、饮食以及药物毒副作用等情况。另取20只荷瘤鼠随机分为4组(分组及处理同上),给药第10天处死小鼠,取肿瘤组织行苏木精-伊红(HE)染色观察肿瘤组织坏死情况并测定肿瘤坏死率。结果人参皂苷Rg3对肿瘤细胞的生长抑制作用随着浓度的升高而逐渐升高,48 h的细胞生长抑制率<24 h的生长抑制率。随着人参皂苷Rg3的浓度升高,小鼠体质量下降越慢;小鼠精神状况、活动、饮食状况等生活质量指标在人参皂苷Rg3组均有明显改善;在整个治疗过程中并未发现人参皂苷Rg3对实验小鼠存在明显毒副作用。对实验小鼠肿瘤组织进行HE染色发现,在人参皂苷Rg3浓度10、20 mg/kg组,肿瘤组织内可见多个小片状坏死,而生理盐水组肿瘤组织无明显组织坏死。生理盐水组、Rg3浓度10 mg/kg组、Rg3浓度20 mg/kg组的肿瘤坏死率分别为20%、60%、80%。结论单药人参皂苷Rg3对BALB/c小鼠大肠癌具有很好的抗肿瘤效果,随着剂量的提高,抗肿瘤效果越明显,是一种剂量依赖型的抗肿瘤药物。

Objective To observe the anticancer efficacy of ginsenoside Rg3 on colorectal cancer in vitro and in vivo. Methods Mice colorectal cell line（CT26） was incubated in 96-well plates（3×103-4×103 per well） with various concentrations of ginsenoside Rg3（0, 5, 10, 15, 20 μg/m L） for 24 hours and 48 hours. 3-（4, 5-dimethyl-2-thiazolyl）-2, 5-dipheny 1-2-H-tetrazolium bromide assay was used to detect the inhibitory rate of cells. Xenograft models were established by subcutaneous implantation of CT26 cells into BABL/c mice. EacThmouse was injected with 1×107 cells suspended in serum-free medium. Xenograft mice were randomized into four groups： physiological saline group, ginsenoside Rg3 5 mg/kg group, ginsenoside Rg3 10 mg/kg group, and ginsenoside Rg3 20 mg/kg group. Ginsenoside Rg3 was administrated to mice by intragastric gavage. All animals were observed for activity, body weight, tumor size, survival time, mental state and adverse ef ect of ginsenoside Rg3. Hematoxylin-eosin stain was used for comparing necrosis rate among groups. Results The inhibitory rates of cells were increasing following the elevating concentrations of ginsenoside Rg3. The anti-proliferation ef ect of ginsenoside Rg3 for 48 hours was weaker than the anti-proliferation ef ect for 24 hours. The decrease of mice body weight was slower than physiological saline group after administration of ginsenoside Rg3, and the number of mice with worse physiological state, lack of activity and loss of appetite in physiological saline group were more than that in ginsenoside Rg3 groups. However, these results among four groups were not significantly dif erent（P 〉 0.05）. There were no obvious adverse ef ects of ginsenoside Rg3 found during the whole study. The necrosis rate of physiological saline group, Rg3 10 mg/kg group and Rg3 20 mg/kg group was 20%, 60% and 80% respectively. Conclusions Ginsenoside Rg3, as a single agent, still has anticancer activity. The anticancer efficacy is increasing following the elevating concentrations of ginsenoside Rg3. Ginsenoside Rg3 is a dose dependent agent.