Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs/Key Laboratory of Agro-Products Processing Technology of Shandong Province/Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, 202 Gongye North Road, Jinan, PR China.
Department of Nephrology, Tai'an City Central Hospital, Tai'an, Shandong, PR China.
J Ethnopharmacol. 2024 Jun 12;327:117835. doi: 10.1016/j.jep.2024.117835. Epub 2024 Mar 14.
The root of Croton crassifolius has been used as a traditional Chinese medicine (TCM), called Radix Croton Crassifolius, and commonly known as "Ji Gu Xiang" in Chinese. Its medicinal value has been recorded in several medical books or handbooks, such as "Sheng Cao Yao Xing Bei Yao", "Ben Cao Qiu Yuan" and "Zhong Hua Ben Cao". It has been traditional employed for treating sore throat, stomach-ache, rheumatism and cancer.
At present, there are limited studies on the evaluation of low-polarity extracts of roots in C. crassifolius. Consequently, the aim of this study was to evaluate the antitumor effect of the low-polarity extract of C. crassifolius root.
Extracts were obtained by supercritical fluid extraction. The extracts were tested for antitumor effects in vitro on several cancer cell lines. A CCK-8 kit was used for further analysis of cell viability. A flow cytometer and propidium iodide staining were used to evaluate the cell cycle and apoptosis. Hoechst staining, JC-1 staining and the fluorescence probe DCFH-DA were used to evaluate apoptotic cells. Molecular mechanisms of action were analyzed by quantitative RT‒PCR and Western blotting. Immunohistochemistry was used for the evaluation of xenograft tumors in male BALB/c mice. Finally, molecular docking was employed to predict the bond between the desired bioactive compound and molecular targets.
Eleven diterpenoids were isolated from low-polarity C. crassifolius root extracts. Among the compounds, chettaphanin II showed the strongest activity (IC = 8.58 μM) against A549 cells. Evaluation of cell viability and the cell cycle showed that Chettaphanin II reduced A549 cell proliferation and induced G2/M-phase arrest. Chttaphanin II significantly induced apoptosis in A549 cells, which was related to the level of apoptosis-related proteins. The growth of tumor tissue was significantly inhibited by chettaphanin II in experiments performed on naked mice. The antitumor mechanism of chettaphanin II is that it can obstruct the mTOR/PI3K/Akt signaling pathway in A549 cells. Molecular docking established that chettaphanin II could bind to the active sites of Bcl-2 and Bax.
Taken together, the natural diterpenoid chettaphanin II was identified as the major antitumor active component, and its potential for developing anticancer therapies was demonstrated for the first time by antiproliferation evaluation in vitro and in vivo.
巴豆的根已被用作中药（TCM），称为巴豆根，中文通常称为“鸡骨香”。其药用价值已被多部医学书籍或手册收录，如《生草药性备要》、《本草求原》和《中华本草》。它传统上被用于治疗喉咙痛、胃痛、风湿和癌症。
目前，对巴豆根的低极性提取物的评价研究有限。因此，本研究的目的是评价巴豆根低极性提取物的抗肿瘤作用。
通过超临界流体萃取获得提取物。将提取物在体外测试对几种癌细胞系的抗肿瘤作用。CCK-8 试剂盒用于进一步分析细胞活力。通过流式细胞仪和碘化丙啶染色评估细胞周期和细胞凋亡。Hoechst 染色、JC-1 染色和荧光探针 DCFH-DA 用于评估凋亡细胞。通过定量 RT-PCR 和 Western blotting 分析作用机制。免疫组织化学用于评价雄性 BALB/c 小鼠异种移植肿瘤。最后，采用分子对接预测所需生物活性化合物与分子靶标的结合。
从巴豆根低极性提取物中分离出 11 种二萜。其中，chettaphanin II 对 A549 细胞的活性最强（IC=8.58 μM）。细胞活力和细胞周期评价表明，Chettaphanin II 可降低 A549 细胞增殖并诱导 G2/M 期阻滞。Chettaphanin II 可显著诱导 A549 细胞凋亡，这与凋亡相关蛋白水平有关。在裸鼠实验中，chettaphanin II 显著抑制肿瘤组织生长。Chettaphanin II 抗肿瘤机制是其可阻断 A549 细胞中的 mTOR/PI3K/Akt 信号通路。分子对接表明 chettaphanin II 可与 Bcl-2 和 Bax 的活性位点结合。
总之，天然二萜 chettaphanin II 被鉴定为主要的抗肿瘤活性成分，其通过体外和体内的增殖评估首次证明了其开发抗癌疗法的潜力。
