解毒三根汤调节肿瘤相关成纤维细胞抗结肠癌侵袭转移及其对α-SMA、Sirt1、Ki67的影响

Jiedu Sangen Decoction Inhibiting Migration and Invasion of Colon Cancer by Regulating the Cancer-associated Fibroblasts, α-SMA, Sirt1 and Ki67

[目的]建立小鼠结肠癌肝转移模型,探讨解毒三根汤通过调节肿瘤相关成纤维细胞(cancer-associated fibroblasts,CAF)抗结肠癌侵袭转移的治疗作用及其对α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、沉默信息调节因子1(silent information regulator 1,Sirt1)、细胞增殖核抗原(nuclcar-associated antigen,Ki67)的影响。[方法]收集对数生长期的结肠癌细胞株(CT-26)和小鼠成纤维细胞(L929),将CT-26细胞及CT-26、L929 1:1混合液分别进行BALB/c雄性小鼠脾被膜下注射,建立结肠癌肝转移模型,分别为模型组和CAF组。造模7d后,将小鼠随机分为空白组、中药组,中药组给予解毒三根汤24g·kg-1灌胃,空白组则予生理盐水20m L·kg-1灌胃,2次/d,连续14d。观察肝、肺脏肿瘤生长情况和转移灶数量,免疫组化检测肝、肺脏的α-SMA、Sirt1、Ki67的表达。[结果]CAF空白组肝脏转移灶数量高于模型空白组,差异有统计学意义(P<0.05);CAF中药组肝脏转移灶数量低于CAF空白组,差异有统计学意义(P<0.05)。肝脏免疫组化结果显示:CAF空白组的α-SMA、Ki67表达量高于模型空白组,差异有统计学意义(P<0.05);CAF中药组的α-SMA、Ki67表达量低于CAF空白组,差异有统计学意义(P<0.05)。CAF空白组的Sirt1表达量低于模型空白组,差异有统计学意义(P<0.05);CAF中药组的Sirt1表达量高于CAF空白组,差异有统计学意义(P<0.05)。肺脏免疫组化结果显示:CAF空白组的α-SMA、Ki67、Sirt1表达量高于模型空白组,差异有统计学意义(P<0.05);CAF中药组的α-SMA、Ki67、Sirt1表达量低于CAF空白组,差异有统计学意义(P<0.05)。[结论]CAF能显著提高肝脏转移灶的数量,解毒三根汤能调控CAF降低肝脏转移灶,降低肝、肺α-SMA、Ki67表达,升高肝脏内Sirt1表达,降低肺脏内Sirt1表达。

Objective]To establish the liver metastasis model of colon cancer in mice, explore the therapeutic effect of Jiedu Sangen Decoction on the invasion and metastasis of colon cancer and the effect of alpha-SMA, Sirt1 and Ki67. [Method] Collect logarithmic growth of CT-26 cells and L929 cells, CT-26 cells and mixture 1：1 respectively for BALB/c male intrasplenic injection, the establishment of liver metastasis model of colon carcinoma, divided into the model group and the cancer-associated fibroblasts group. BALB/c male mice were randomly divided into the blank group and the Chinese medicine group, 0.4mL·20g-1 intragastric administration, the concentration of Jiedu Sangen Decoction was 1.2g·mL-1, while the blank group was given physiological saline. Give the medicine two times a day, and give the medicine 14 days. The alpha-SMA, Sirt and Ki67 of liver and lung were detected by immunohistochemistry. [Result] The number of liver metastasis in the CAF blank group was significantly increased compared with that in the model blank group（P〈0.05）. The liver metastasis and liver quality were significantly decreased dcompared with the CAF blank group（P〈0.05）. Liver immunohistochemistry showed that the expression of alpha-SMA and Ki67 in the CAF blank group was significantly higher than that in the model blank group（P〈0.05）, and CAF TCM group was significantly lower in the CAF blank group（P〈0.05）. The expression of Sirt1 was significantly lower in CAF blank group than in model blank group（P〈0.05）, and significantly higher in the CAF TCM group than that in the CAF blank group（P〈0.05）. The CAF blank group＇s expression of alpha-SMA, Ki67 and Sirt1 in lung was significantly higher than that in the model blank group（P〈0.01）, and the CAF TCM group was significantly lower than that in the CAF blank group（P〈0.05）. [Conclusion] CAF can significantly improve the number of liver metastasis. Jiedu Sangen Decoction can reduce the expression of alpha-SMA, Ki67, and increase the expression of Sirt1 in the liver, and reduce the expression of Sirt1 in the lung.