虎杖苷对THP-1细胞增殖及凋亡的影响及其作用机制研究

Effect of polydatin on the proliferation and apoptosis of THP-1 cells and the mechanism

摘要:

目的 探讨虎杖苷对急性单核细胞白血病细胞株THP-1增殖及凋亡的影响及可能的作用机制。 方法 以不同梯度浓度的虎杖苷处理THP-1细胞24 h、48 h，CCK-8法检测细胞增殖活力，并计算半数抑制浓度。取对数生长期的THP-1细胞，分为虎杖苷处理组（处理浓度为半数抑制浓度）和空白对照组（细胞中未施加虎杖苷溶液处理），培养48 h后，采用流式细胞术检测细胞凋亡及细胞周期；Western blot法检测PI3K、AKT、p-AKT、mTOR、p-mTOR、p70 S6K、p-p70 S6K蛋白的表达。 结果 虎杖苷可有效抑制THP-1细胞的增殖，48 h的半数抑制浓度为1 800 μmol/L。经1 800 μmol/L的虎杖苷溶液作用48 h后，THP-1细胞的凋亡率较空白对照组显著增加（P<0.05）；细胞周期出现G₀/G₁期至S期的阻滞，表现为G₀/G₁期的细胞比例较空白对照组明显上升，S期的细胞比例较空白对照组显著下降（P<0.05）；PI3K、AKT、p-AKT、mTOR、p-mTOR、p70 S6K、p-p70 S6K蛋白表达较空白对照组显著降低（P<0.05）。 结论 虎杖苷可有效抑制THP-1细胞的增殖，阻滞细胞周期并诱导细胞凋亡，其作用机制可能与PI3K/AKT/mTOR信号通路的抑制表达有关。

Abstract:

Objective To explore the effect of polydatin on the proliferation and apoptosis of acute monocytic leukemia cell line THP-1 and the possible mechanism. Methods After THP-1 cells were treated with polydatin at gradient concentrations for 24 hours and 48 hours, their proliferation was determined by CCK-8 assay, and half maximal inhibitory concentration (IC50) was calculated. Logarithmically growing THP-1 cells were divided into two groups, a polydatin treatment group (treated with IC50 of polydatin) and a blank control group (treated without polydatin solution), and incubated for 48 hours. Cell apoptosis and cell cycle were measured by flow cytometry. The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins were measured by Western blotting. Results After treatment with polydatin, the proliferation of THP-1 cells was strongly inhibited, and the IC50 at 48 hours was 1 800 μmol/L. After treatment with 1 800 μmol/L polydatin solution for 48 hours, the apoptosis rate of THP-1 cells increased significantly compared with the blank control group (P<0.05). The cell cycle was arrested in the G0/G1 and S phases, with a significantly increased proportion of cells in the G0/G1 phase and a significantly decreased proportion of cells in the S phase, as compared with the blank control group (P<0.05). The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins decreased significantly compared with the blank control group (P<0.05). Conclusions Polydatin can effectively inhibit the proliferation, block the cell cycle, and induce the apoptosis of THP-1 cells, which may be related to inhibition of the PI3K/AKT/mTOR signaling pathway.