Department of Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia.
Department of Pharmaceutical Biology, Faculty of Pharmacy, Widya Mandala Catholic University, Surabaya, Indonesia.
Asian Pac J Cancer Prev. 2021 Jun 1;22(6):1967-1973. doi: 10.31557/APJCP.2021.22.6.1967.
Nature has provided us with many pharmaceutical resources so far. Breast cancer shows an increasing trend in the world for the last decade and becomes one of five leading causes of death. Among the plants, Melia azedarach L. has been used widely in traditional medicine for many ailments including breast cancer. Following our previous findings that the ethyl acetate fraction was the most active cytotoxic fraction against T47D cells, we aimed to isolate the cytotoxic compounds and further elucidate their apoptotic mechanisms.
The compounds were isolated through a series of chromatography with cytotoxicity evaluations. Identification of the isolated compounds was achieved by intensive spectroscopic analysis such as NMR, MS, and IR spectra. Cytotoxicity was evaluated by MTT method using doxorubicin as a reference compound. The expression of apoptosis-related factors was quantified by flow cytometry and immunocytochemistry.
Two isomers of pregnane steroids with molecular weight 330.2087 (C21H30O3) were isolated from the EtOAc extract. Spectroscopic analysis revealed the structures as 17-ethylene-3,4-dihydroxy-14-methyl-18-norandrostene-16-one (1) and 17-ethylene-3,4-dihydroxy-5-pregnene-16-one (2), respectively. These compounds showed moderate cytotoxicity (IC50 172.9 and 62.2 µg/mL, respectively) comparable to doxorubicin (IC50 3.08 µg/mL). The execution of apoptosis may be related to the increase of the ratio of BAX/bcl-2 of the cells. Conclusion: The EtOAc fraction of Melia azedarach L. leaves and the isolated 5-pregnene-16-one steroids are promising reagents for breast cancer treatment by introducing apoptosis to tumor cells. However, further researches are required to highlight its safety and usage in vivo.
.
大自然迄今为止为我们提供了许多药物资源。在过去的十年中，乳腺癌在全球呈上升趋势，成为导致死亡的五个主要原因之一。在植物中，苦楝已被广泛用于传统医学治疗多种疾病，包括乳腺癌。根据我们之前的研究结果，乙酸乙酯部分是对 T47D 细胞最具细胞毒性的部分，我们旨在分离具有细胞毒性的化合物，并进一步阐明其凋亡机制。
通过一系列具有细胞毒性评估的色谱法分离化合物。通过 NMR、MS 和 IR 光谱等综合光谱分析鉴定分离出的化合物。通过 MTT 法评价细胞毒性，以阿霉素为参考化合物。通过流式细胞术和免疫细胞化学定量检测凋亡相关因子的表达。
从 EtOAc 提取物中分离出两种具有分子量 330.2087（C21H30O3）的孕甾烷类异构体。光谱分析表明结构分别为 17-乙烯基-3,4-二羟基-14-甲基-18-降孕烯-16-酮（1）和 17-乙烯基-3,4-二羟基-5-孕烯-16-酮（2）。这些化合物表现出中等的细胞毒性（IC50 分别为 172.9 和 62.2 µg/mL，与阿霉素（IC50 为 3.08 µg/mL）相当。细胞凋亡的执行可能与细胞中 BAX/bcl-2 比值的增加有关。
苦楝叶的乙酸乙酯部分和分离出的 5-孕烯-16-酮甾体是通过诱导肿瘤细胞凋亡治疗乳腺癌的有前途的试剂。然而，需要进一步的研究来强调其安全性和体内应用。
