Luzhou City Hospital of Traditional Chinese Medicine, Luzhou, China.
Luzhou City Hospital of Traditional Chinese Medicine, Luzhou, China..
Life Sci. 2022 Jan 1;288:118993. doi: 10.1016/j.lfs.2020.118993. Epub 2021 Feb 2.
To study the proapoptotic effect of ligustilide on osteoblastoma (OS) and the relative related molecular mechanism.
An MTT was used to examine the proliferation of OS cells, and Flow cytometry was used to analyze apoptosis and the cell cycle. Western blotting was used to detect the signaling pathway of apoptosis, and immunohistochemical (IH) staining was used to detect the apoptosis status of OS cells. A TLR4 inhibitor was used to study the effect of ligustilide on OS.
Ligustilide inhibited OS cell proliferation but had no inhibitory effect on normal bone marrow cells. Flow cytometry results showed that ligustilide induced apoptosis in OS cells, and the cell cycle was arrested at the M/G2 phase. Western blot results showed that ERK, P53, P21, Caspase 9, Caspase 8 and Caspase 3 were all activated; cytochrome C and Bax increased; and Bcl-2 decreased when OS was treated with ligustilide. When an ERK or Caspase inhibitor was added to the culture medium, the apoptosis of OS cells decreased to some degree. When OS cells were pretreated with CLI-095, which is a TLR4 inhibitor, the percentage of apoptotic cells and cell cycle arrest were both reversed. IH results also showed that ligustilide induced apoptosis in OS cells, and the effect was blocked by the TLR4 inhibitor.
Ligustilide selectively inhibited the proliferation of OS cells by inducing apoptosis, which possibly included endogenous and exogenous apoptosis through TLR4.
研究蛇床子素对成骨细胞瘤（OS）的促凋亡作用及其相关分子机制。
采用 MTT 法检测 OS 细胞的增殖，流式细胞术分析细胞凋亡和细胞周期，Western blot 检测凋亡相关信号通路，免疫组化（IH）染色检测 OS 细胞的凋亡状态，采用 TLR4 抑制剂研究蛇床子素对 OS 的作用。
蛇床子素抑制 OS 细胞增殖，但对正常骨髓细胞无抑制作用。流式细胞术结果显示，蛇床子素诱导 OS 细胞凋亡，细胞周期阻滞在 M/G2 期。Western blot 结果显示，蛇床子素处理 OS 后 ERK、P53、P21、Caspase 9、Caspase 8 和 Caspase 3 均被激活，细胞色素 C 和 Bax 增加，Bcl-2 减少。当向培养基中加入 ERK 或 Caspase 抑制剂时，OS 细胞的凋亡程度在一定程度上降低。当 OS 细胞预先用 TLR4 抑制剂 CLI-095 处理时，凋亡细胞的百分比和细胞周期阻滞均被逆转。IH 结果也表明蛇床子素诱导 OS 细胞凋亡，该作用被 TLR4 抑制剂阻断。
蛇床子素通过诱导凋亡选择性抑制 OS 细胞的增殖，其可能通过 TLR4 途径包括内源性和外源性凋亡。
