Longping Branch Graduate School, Hunan University, Changsha 410082, China.
School of Public Health, Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing 100069, China.
Molecules. 2020 Feb 20;25(4):946. doi: 10.3390/molecules25040946.
5,6,7,8,3,4'-Hexamethoxyflavone, also called nobiletin (NOB), widely found in the citrus peel, is one of the main byproducts in citrus processing. NOB is considered safe, but its safety for women during pregnancy is unknown. Therefore, the effect of NOB on apoptosis in human choriocarcinoma trophoblast cells (BeWo cells) was evaluated. Cells were divided into four groups and cultured with different concentrations of NOB (0, 10, 33, and 100 μM) for 12, 24, 36, and 48 h respectively. Cell viability was detected by CCK-8 assay, cell morphology was detected by a Cell Imaging Multi-Mode Reader, and cell cycle and apoptosis were detected by flow cytometry. Cleaved PARP level, the expressions of B cell lymphoma 2 (BCL2) family proteins, and p53 pathway proteins were detected by Western blot. The results showed that after 48 h of cell culture, the cell viability was decreased significantly, but apoptosis was significantly increased. Compared to the cells without NOB treatment, the cells treated with NOB at 10 or 33 μΜ showed no significant differences in the number of suspended cells or late apoptosis rate, except the increase of cell viability. Treatment of NOB at the concentration of 100 μM improved cell viability, attenuated apoptosis, decreased suspended cells, and did not alter the G1 phase arrest, compared with the non-NOB-treated group after 48 h of culturing. The 100 μΜ NOB treatment increased the levels of BCL2 and BCLX, and decreased p53 accumulation in BeWo cells at 48 h, but had no effect on the expression of BAX, BAK, BAD, p21, and G1 phase arrest. These findings provide evidence that NOB (10, 33, and 100 μΜ) was safe for BeWo cells. NOB at the concentration of 100 μΜ could attenuate apoptosis in BeWo cells, which might be helpful to prevent pregnancy-related diseases caused by apoptosis.
5,6,7,8,3,4'-六甲氧基黄酮，也称为诺必灵（NOB），广泛存在于柑橘皮中，是柑橘加工的主要副产物之一。NOB 被认为是安全的，但它在孕妇中的安全性尚不清楚。因此，评估了 NOB 对人绒毛膜癌细胞（BeWo 细胞）凋亡的影响。细胞分为四组，分别用不同浓度的 NOB（0、10、33 和 100μM）培养 12、24、36 和 48 小时。通过 CCK-8 检测细胞活力，通过细胞成像多模式读取器检测细胞形态，通过流式细胞术检测细胞周期和凋亡。通过 Western blot 检测裂解 PARP 水平、BCL2 家族蛋白和 p53 通路蛋白的表达。结果表明，细胞培养 48 小时后，细胞活力明显下降，但凋亡明显增加。与未用 NOB 处理的细胞相比，用 10 或 33μM 的 NOB 处理的细胞悬浮细胞数量或晚期凋亡率无显著差异，除细胞活力增加外。与未用 NOB 处理的细胞相比，100μM 的 NOB 处理在 48 小时培养后可提高细胞活力，减轻凋亡，减少悬浮细胞，且不改变 G1 期阻滞，与未用 NOB 处理的细胞相比。100μM 的 NOB 处理增加了 BeWo 细胞中 BCL2 和 BCLX 的水平，并减少了 p53 的积累，但对 BAX、BAK、BAD、p21 的表达和 G1 期阻滞没有影响。这些发现为 10、33 和 100μM 的 NOB 对 BeWo 细胞安全提供了证据。100μM 的 NOB 可以减轻 BeWo 细胞的凋亡，这可能有助于预防由凋亡引起的妊娠相关疾病。
