黄芪-莪术配伍对结肠癌原位移植瘤小鼠模型抗结肠癌生长转移的干预效应研究

Study on intervention effect of Astragali Radix-Curcumae Rhizoma on growth and metastasis of colon cancer in orthotopic transplantation mice model of colon cancer

黄芪-莪术药对是临床治疗肿瘤的常用组合,主要用于消化道相关炎症及肿瘤的治疗,临床所用配比也多不固定。为研究该药对在不同配比下抗肿瘤效果是否存在差异,该研究建立结肠癌原位移植瘤小鼠模型,结合主成分分析和聚类分析探讨药对不同配比对结肠癌小鼠肿瘤生长、转移的作用,优选黄芪-莪术抗结肠癌效应最适配比。给药15 d后测量肿瘤小鼠除瘤体质量、肿瘤体积、肝转移灶数目,HE染色法观察肿瘤组织和肝脏组织病理学改变,Western blot法检测荷瘤小鼠肿瘤组织中肿瘤生长相关指标:细胞核相关抗原Ki-67(Ki67)、HMG盒转录因子1(HMG-box transcription factor 1, HBP1)、甲胎蛋白(alpha-fetoprotein, AFP)和肝脏中肿瘤转移相关指标:β-连环蛋白(β-catenin)、上皮型钙黏附素(E-cadherin)、波形蛋白(vimentin)、蛋白质53(protein53, p53)的蛋白表达水平,随后采用主成分分析和聚类分析法优选黄芪-莪术抗结肠癌效应最适配比。实验结果表明,不同配比黄芪-莪术不同程度抑制了肿瘤的生长和肝转移,黄芪-莪术1∶1配比组对肿瘤生长抑制作用最优,2∶1配比组抑制肿瘤肝转移和改善肿瘤小鼠体质量效果更优。黄芪-莪术配伍显著上调结肠癌小鼠肿瘤组织中HBP1蛋白表达,显著下调肿瘤组织中Ki67和AFP蛋白表达;同时黄芪-莪术配伍显著上调结肠癌小鼠肝脏组织中E-cadherin蛋白表达,显著降低β-catenin, vimentin和p53蛋白表达水平。主成分分析结果显示黄芪-莪术配伍组距假手术组较近的前3组依次为2∶1,1∶1,3∶2组,其中2∶1组中心距离距假手术组最短,提示黄芪-莪术2∶1组对小鼠结肠癌干预作用最优,与临床常用配比相一致。聚类结果将11个组别按黄芪-莪术配伍组、假手术组、模型组聚为3类,与理论实际相符合。该研究结果为临床更有效应用黄芪-莪术治疗结肠癌提供了依据,为经典药对的开发提供新的思路。

Astragali Radix-Curcumae Rhizoma is a classic drug pair mainly used for the treatment of digestive tract-related inflammation and tumors, but the ratio is not fixed in clinical practice. In order to study whether the anti-tumor effect of the drug pair is diffe-rent under different ratios, orthotopic transplantation model of colon cancer was established in mice. Then the principal component analysis(PCA) and cluster analysis(CA) were used to explore the effect of different ratios of the drug pair on the tumor growth and metastasis, and select the optimal ratio of Astragali Radix-Curcumae Rhizoma for anti-colon cancer effect. After administration for 15 days, the body weight of colon cancer mice with the tumor removed, the tumor volume and the number of liver metastases were mea-sured;the pathological changes of tumor tissue and liver tissue were observed by HE staining. At the same time, Western blot method was used to detect the protein expression level of tumor growth-related indicators in tumor tissue(Ki67, HBP1, AFP) and tumor metastasis-related indicators in liver tissue(β-catenin, E-cadherin, vimentin, p53) of the tumor-bearing mice. Subsequently, PCA and CA were used to select the optimal ratio of Astragali Radix-Curcumae Rhizoma for anti-colon cancer effect. The experimental results showed that different ratios of Astragali Radix-Curcumae Rhizoma inhibited tumor growth and metastasis to varying degrees. The ratio at 1∶1 of Astragali Radix-Curcumae Rhizoma had the best inhibitory effect on tumor growth, and the 2∶1 ratio group had the best effect on inhibiting liver metastasis and improving weighed loss. Astragali Radix-Curcumae Rhizoma significantly up-regulated the protein expression of HBP1 in tumor tissue of colon cancer mice, and significantly down-regulated the protein expression of Ki67 and AFP in tumor tissue;meanwhile, Astragali Radix-Curcumae Rhizoma significantly up-regulated the protein expression of E-cadherin in liver tissue of colon cancer mice, and significantly reduced the protein expression of β-catenin, vimentin and p53 in liver tissue. PCA results showed that the first three groups in the Astragali Radix-Curcumae Rhizoma compatibility group that were closer to the sham operation group were in the order of 2∶1, 1∶1 and 3∶2, among which the center distance of the 2∶1 group was the shortest from the sham operation group, indicating that the ratio 2∶1 of Astragali Radix-Curcumae Rhizoma had the best intervention effect on colon cancer in mice, consistent with the commonly used clinical proportion. CA results showed that 11 groups of colon cancer mice were classified into 3 categories: Astragali Radix-Curcumae Rhizoma compatibility group, sham operation group and model group, which was consistent with the theory. The results of this study provide a basis for more effective clinical application of Astragali Radix-Curcumae Rhizoma in the treatment of colon cancer, and provide new ideas for the development of classic drug pairs.