散结抗瘤方体内和体外抑瘤作用的实验研究

In vivo and in vitro anti-tumor effect of Sanjie Kangliu Decoction

目的探讨散结抗瘤方体内和体外抗肿瘤的可能机制。方法体内实验:对荷瘤小鼠(SRS-82瘤株)用散结抗瘤方煎剂灌胃,计算抑瘤率,电镜下观察肿瘤细胞形态,并提取肿瘤细胞DNA做琼脂糖凝胶电泳。体外实验:传代培养 SRS-82细胞,用散结抗瘤方含药血清干预细胞后,检测活细胞数、细胞凋亡率、Bcl-2蛋白MMP-2mRNA的表达强度。结果与空白对照组比较,散结抗瘤方组瘤重显著减轻,电镜下可见肿瘤细胞出现凋亡特征性改变;细胞活力明显降低, 细胞凋亡率明显升高,细胞Bcl-2蛋白的表达强度降低。结论散结抗瘤方在体内有明显的抑瘤作用。在体外对肿瘤细胞也有明显的抑制增殖作用。该方抑瘤作用可能与其诱导肿瘤细胞凋亡密切相关,机制可能与下调抑制凋亡基因Bcl-2 的表达有关。散结抗瘤方对肿瘤细胞MMP-2的表达没有影响。

Objective To study the anti-tumor effect ofSanjie Kangliu （swelling-dispersing） Decoction in vivo and in vitro. Methods The mice bearing tumors （established by subcutaneous inoculation of SRS-82 cells） were fed with Sanjie Kangliu Decoction. The tumor was isolated and weighed to calculate the tumor inhibition rate, followed by morphological observation of the tumor cells under electron microscope. The DNA of the tumor cells was then extracted for agarose gel electrophoresis. In the in vitro experiment, cultured SRS-82 cells were treated with the serum of rats fed with Sanjie Kangliu Decoction, and the cell viability and apoptosis was examined by MTT assay and flow cytometry, respectively. Immunocytochemistry was employed to test the expression intensity of Bcl-2 protein of the rat serum-treated cells, and in situ hybridization performed to detect matrix metalloproteinase （MMP）-2 mRNA expression. Results In the tumor-bearing mouse models, Sanjie Kangliu Decoction administration resulted in significant reduction of the tumor mass and apoptosis of the tumor cells as shown by typical apoptotic changes of the cell morphology and presence of DNA ladder in agarose gel electrophoresis. In comparison with the mice without the decoction treatment, the treated mice showed significantly lower tumor cell viability, higher cell apoptosis rate, and weaker Bcl-2 expression. The expression of MMP-2 mRNA showed no distinct difference between the groups. Conclusion Sanjie Kangliu Decoction has obvious anti-tumor effect both in vivo and in vitro possibly through the mechanism of inducing tumor cell apoptosis by reducing Bcl-2 expression without affecting the expression of MMP-2 mRNA.