疏肝健脾解毒方对乳腺癌癌前病变防治作用机制研究

Preventive and Therapeutic Mechanism of Shugan Jianpi Jiedu Decoction on Precancerous Lesions of Breast Cancer

关键词:  疏肝健脾解毒方  乳腺癌癌前病变  雌激素受体  PI3K/Akt/mTOR信号通路  免疫应答

Key words:  Shugan Jianpi Jiedu decoction  precancerous lesions of breast cancer  estrogen receptor  PI3K/Akt/mTOR signaling pathway  escape of immunity

摘要:

目的 通过动物实验研究疏肝健脾解毒方对乳腺癌癌前病变的作用机制。方法 取SD大鼠，分为6组(每组10只)：空白组、乳腺癌癌前病变模型组、他莫昔芬组和疏肝健脾解毒方低、中、高剂量组。采用二甲基苯蒽造模法对乳腺癌癌前病变大鼠模型造模。采用HE染色观察各组乳腺组织病理变化，流式细胞仪检测CD4⁺、CD8⁺含量；ELISA检测IL-2、IL-4、IL-6、IL-10、IL-12、E2、P含量。Western blotting检测ER、PI3K、p-Akt及mTOR的蛋白表达。结果 HE染色显示大鼠乳腺组织发生改变，提示造模成功。与空白组比较，模型组外周血中CD4⁺含量降低，CD8⁺含量升高，差异有统计学意义(P<0.01)；与模型组比较，疏肝健脾解毒方低、中、高剂量组和他莫昔芬组CD4⁺含量增加，CD8⁺含量下降，差异均有统计学意义(P<0.01)。与空白组比较，模型组IL-2、IL-4、IL-10浓度显著降低(P<0.01)，IL-12、IL-6显著升高(P<0.01)；与模型组比较，疏肝健脾解毒方低、中、高剂量组和他莫昔芬组IL-2、IL-4、IL-10浓度均显著升高(P<0.01)，IL-12、IL-6显著下降(P<0.05或P<0.01)。与空白组比较，模型组E2和P含量显著升高(P<0.01)；与模型组比较，疏肝健脾解毒方低、中、高剂量组和他莫昔芬组E2和P含量均显著降低(P<0.01)。Western blotting检测结果表明，与空白组比较，模型组ER、PI3K、p-Akt及mTOR的表达显著升高(P<0.01)；与模型组比较，疏肝健脾解毒方低、中、高剂量组和他莫昔芬组ER、PI3K、p-Akt及mTOR的表达均显著下降(P<0.01)。结论 疏肝健脾解毒方可能是通过使ER的表达下降，从而抑制PI3K/Akt/mTOR信号通路的表达，同时影响机体免疫应答来逆转大鼠乳腺癌变进程。

Abstract:

OBJECTIVE To study the efficacy and mechanism of Shugan Jianpi Jiedu decoction in the treatment of the precancerous lesions of breast cancer through animal experiment. METHODS SD rats were taken and divided into 6 groups(10 rats in each group), namely blank group, breast precancerous lesion model group, tamoxifen group, Shugan Jianpi Jiedu decoction groups with low dose, middle dose, and high dose. DMBA modeling method was used to carry out modeling for breast precancerous lesion. HE staining was used to observe the pathological changes of breast tissue. CD4⁺, CD8⁺ were detected by flow cytometry. ELISA was used to detect IL-2, IL-4, IL-6, IL-10, IL-12, E2, P. The protein expression of ER, PI3K, p-Akt and mTOR was detected by Western blotting. RESULTS HE staining showed changes in rat mammary tissue, indicating successful modeling. Compared with the blank group, the content of CD4⁺ decreased and the content of CD8⁺ increased in the model group(P<0.01); compared with model group, the content of CD4⁺ increased and the content of CD8⁺ decreased in low, middle, high dose groups of Shugan Jianpi Jiedu decoction and tamoxifen group(P<0.01). The levels of IL-2, IL-4 and IL-10 in the model group were significantly lower than those in the blank group(P<0.01), while IL-12 and IL-6 were significantly increased(P<0.01). Compared with the model group, the concentrations of IL-2, IL-4 and IL-10 in the low, middle and high dose groups of Shugan Jianpi Jiedu decoction and the tamoxifen group were significantly increased(P<0.01), while IL-12 and IL-6 decreased significantly(P<0.05 or P<0.01). Compared with the blank group, the contents of E2 and P in the model group increased significantly(P<0.05 or P<0.01), the contents of E2 and P in the low, middle and high dose groups of Shugan Jianpi Jiedu decoction and the tamoxifen group were significantly lower than those in the model group(P<0.01). The Western blotting results showed that compared with the blank group, the expression of ER, PI3K, p-Akt and mTOR in the model group was significantly increased(P<0.01). Compared with the model group, the expression of ER, PI3K, p-Akt and mTOR in the low, medium and high dose groups of Shugan Jianpi Jiedu decoction and the tamoxifen group were significantly decreased(P<0.01). CONCLUSION Shugan Jianpi Jiedu decoction may inhibit the expression of ER, thus inhibiting the expression of PI3K/Akt/mTOR signaling pathway. Meanwhile, it can affect the immune response and reverse the precancerous lesions of breast cancer.