State Key Laboratory of Southwestern Chinese Medicine Resources, Pharmacy School, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
National Cancer Center, National Clinical Research Center for Cancer, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China.
J Ethnopharmacol. 2022 Jun 12;291:115104. doi: 10.1016/j.jep.2022.115104. Epub 2022 Feb 24.
Xiaojin Pills was first recorded during the Qing Dynasty and have a history of nearly 300 years. It is the first choice among Chinese patent medicines for the clinical treatment of diseases of the mammary glands in contemporary traditional Chinese medicine. It was also widely used in the treatment of breast cancer, lung cancer, thyroid cancer, and other malignant tumors. Its initial administration method was "taken orally after soaking with Chinese baijiu"; however, the method was changed to "taken orally with water" within the last 40 years. There is no scientific evidence for the difference in efficacy against breast cancer between the two methods of administration.
In vitro and in vivo experiments were carried out to confirm the therapeutic advantages of the liquor extract of Xiaojin Pills to improve the efficacy against breast cancer, and the mechanism was explained in terms of metabolomics and molecular biology.
In vitro, a cell counting kit-8 cell activity assay and flow cytometry were used to detect the activity and apoptosis of MCF-7 breast cancer cells. In vivo, pharmacodynamic evaluation was performed by constructing a heterotopic transplantation model of breast cancer in BALB/c-nu mice. TUNEL staining was used to observe the apoptosis of cells in tumor tissues. The expression of proteins associated with the phosphoinositide 3-kinase (PI3K)/Akt pathway in BALB/c-nu mice tissue was investigated by metabolomics analysis, immunohistochemistry and western blotting.
CCK-8 assay showed that the IC of XJP-L for the inhibition of the activity of MCF-7 cells was less than that of XJP-W at different times. Flow cytometry assay suggested that the apoptosis rate in the XJP-L group was higher than that in the normal control group (p < 0.01). Animal experiment results indicated that both XJP-W group and XJP-L group reduced the volume and quality of the tumor after administration, and the reduction was more significant in the XJP-L group (p < 0.01). Metabolomics analysis results demonstrated that there are about 26 different metabolites have been screened in the serum metabolites between the liquor and water extract, mainly involved in glycerophospholipid, glutamic acid, aspartic acid, nitrogen and pyrimidine metabolism. In addition, immunohistochemistry and WB results showed that compared with the model group, the protein expression of PTEN, AKT, BAX and in tumor tissues of XJP-L and XJP-W groups both exhibited an upward trend, while the expression of BCL-2, p-PI3K and p-AKT exhibited a downward trend, which was much more obvious in XJP-L group.
This study demonstrates that the liquor extract of Xiaojin Pills had a stronger anti-breast cancer effect than that of the water extract. The PI3K/Akt signaling pathway might play an important role in the mechanism of the liquor extract of Xiaojin Pills and thus improve the efficacy against breast cancer.
小金丸最早记载于清代，已有近 300 年的历史。在当代中医临床治疗乳腺疾病中，它是中药成药的首选。它也被广泛用于治疗乳腺癌、肺癌、甲状腺癌等恶性肿瘤。其最初的给药方法是“用中国白酒浸泡后口服”；然而，在过去的 40 年中，给药方法改为“用水口服”。两种给药方法对乳腺癌的疗效没有科学证据。
通过体外和体内实验，证实小金丸酒提物在提高乳腺癌疗效方面的治疗优势，并从代谢组学和分子生物学角度解释其机制。
体外采用细胞计数试剂盒-8 细胞活性测定和流式细胞术检测 MCF-7 乳腺癌细胞的活性和凋亡。体内，通过构建 BALB/c-nu 小鼠乳腺癌异位移植模型进行药效学评价。TUNEL 染色观察肿瘤组织中细胞的凋亡。通过代谢组学分析、免疫组化和 Western blot 研究 BALB/c-nu 小鼠组织中与磷酸肌醇 3-激酶（PI3K）/Akt 通路相关的蛋白表达。
CCK-8 检测结果表明，XJP-L 对 MCF-7 细胞活性的抑制 IC 在不同时间均小于 XJP-W。流式细胞术检测结果表明，XJP-L 组细胞凋亡率明显高于正常对照组（p<0.01）。动物实验结果表明，XJP-W 组和 XJP-L 组给药后均可减少肿瘤体积和质量，XJP-L 组减少更为显著（p<0.01）。代谢组学分析结果表明，在血清代谢物中，酒提物和水提物之间筛选出约 26 种不同的代谢物，主要涉及甘油磷脂、谷氨酸、天冬氨酸、氮和嘧啶代谢。此外，免疫组化和 WB 结果表明，与模型组相比，XJP-L 和 XJP-W 组肿瘤组织中 PTEN、AKT、BAX 蛋白表达呈上升趋势，BCL-2、p-PI3K、p-AKT 表达呈下降趋势，XJP-L 组更为明显。
本研究表明小金丸酒提物的抗乳腺癌作用强于水提物。PI3K/Akt 信号通路可能在小金丸酒提物的作用机制中发挥重要作用，从而提高乳腺癌的疗效。
