The First Affiliated Hospital of Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, PR China; Shandong Qingdao No. 2 Health School, Qingdao, PR China.
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, PR China.
Biomed Pharmacother. 2021 Feb;134:111122. doi: 10.1016/j.biopha.2020.111122. Epub 2020 Dec 16.
Brucea javanica oil (BJO), one of the main products of Brucea javanica, has been widely used in treating different kinds of malignant tumors. Quassinoids are the major category of anticancer phytochemicals of B. javanica. However, current researches on the anti-cancer effect of BJO mainly focused on oleic acid and linoleic acid, the common major components of dietary edible oils, essential and characteristic components of B. javanica like quassinoids potentially involved remained unexplored. In the current investigation, we developed an efficient HPLC method to detect brusatol, a characteristic quassinoid, and comparatively scrutinized the anti-hepatocellular carcinoma (anti-HCC) effect of BJO, brusatol-free BJO (BF-BJO), and brusatol-enriched BJO (BE-BJO) against hepatoma 22 (H22) in mice. High-performance liquid chromatography (HPLC) was utilized to identify the components in BJO. BE-BJO was extracted with 95 % ethanol. The anti-tumor effect of BJO, BF-BJO and BE-BJO was comparatively investigated, and the potential underlying mechanism was explored in H22 ascites tumor-bearing mice. The results indicated that BJO and BE-BJO significantly prolonged the survival time of H22 ascites tumor-bearing mice, while BF-BJO exhibited no obvious effect. BJO and BE-BJO exhibited pronounced anti-HCC activity by suppressing the growth of implanted hepatoma H22 in mice, including ascending weight, abdominal circumference, ascites volume and cancer cell viability, with a relatively wide margin of safety. BJO and BE-BJO significantly induced H22 cell apoptosis by upregulating the miRNA-29b gene level and p53 expression. Furthermore, BJO and BE-BJO treatment substantially downregulated Bcl-2 and mitochondrial Cytochrome C protein expression, and upregulated expression levels of Bax, Bad, cytosol Cytochrome C, caspase-3 (cleaved), caspase‑9 (cleaved), PARP and PARP (cleaved) to induce H22 cells apoptosis. Brusatol was detected in BJO and found to be one of its major active anti-HCC components, rather than fatty acids including oleic acid and linoleic acid. The anti-HCC effect of BJO and BE-BJO was intimately associated with the activation of miRNA-29b, p53-associated apoptosis and mitochondrial-related pathways. Our study gained novel insight into the material basis of BJO in the treatment of HCC, and laid a foundation for a novel specific standard for the quality evaluation of BJO and its commercial products in terms of its anti-cancer application.
鸦胆子油（BJO）是鸦胆子的主要产品之一，已广泛用于治疗各种恶性肿瘤。三萜类化合物是鸦胆子抗癌植物化学物质的主要类别。然而，目前关于 BJO 的抗癌作用的研究主要集中在油酸和亚油酸上，而鸦胆子的必需和特征成分，如三萜类化合物，尚未得到充分研究。在本研究中，我们开发了一种高效液相色谱法（HPLC）来检测鸦胆子苦醇，这是一种特征性的三萜类化合物，并比较了 BJO、不含鸦胆子苦醇的 BJO（BF-BJO）和富含鸦胆子苦醇的 BJO（BE-BJO）对小鼠肝癌 22（H22）的抗肝癌（抗 HCC）作用。高效液相色谱法（HPLC）用于鉴定 BJO 中的成分。BE-BJO 用 95%乙醇提取。比较了 BJO、BF-BJO 和 BE-BJO 的抗肿瘤作用，并在 H22 腹水瘤荷瘤小鼠中探讨了其潜在的作用机制。结果表明，BJO 和 BE-BJO 显著延长了 H22 腹水瘤荷瘤小鼠的生存时间，而 BF-BJO 则无明显作用。BJO 和 BE-BJO 通过抑制植入的肝癌 H22 在小鼠中的生长，表现出明显的抗 HCC 活性，包括体重上升、腹围增大、腹水体积减少和癌细胞活力降低，且具有较宽的安全范围。BJO 和 BE-BJO 通过上调 miRNA-29b 基因水平和 p53 表达，显著诱导 H22 细胞凋亡。此外，BJO 和 BE-BJO 处理可显著下调 Bcl-2 和线粒体细胞色素 C 蛋白表达，上调 Bax、Bad、细胞质细胞色素 C、caspase-3（切割）、caspase-9（切割）、PARP 和 PARP（切割）表达，诱导 H22 细胞凋亡。在 BJO 中检测到鸦胆子苦醇，发现其是其主要的活性抗 HCC 成分之一，而不是油酸和亚油酸等脂肪酸。BJO 和 BE-BJO 的抗 HCC 作用与 miRNA-29b 的激活、p53 相关凋亡和线粒体相关途径密切相关。本研究为鸦胆子油治疗 HCC 的物质基础提供了新的见解，并为鸦胆子油及其商业产品的抗癌应用质量评价建立了一种新的特异性标准奠定了基础。
