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目的探讨板蓝根活性部位对Hep-2、Hela、U937、Jukart细胞的细胞凋亡和小鼠单核巨噬细胞白血病细胞RAW264.7表达细胞因子IL-10、TNF-α的影响。方法 0.5 g/L板蓝根活性部位作用于Hep-2、Hela、U937、Jukart 24 h,经PI染色,流式细胞术研究其对细胞凋亡的影响;100TCID50的单纯疱疹病毒(HSV-1)作用于U937,0.5 g/L药物处理24 h,流式细胞术研究其对细胞凋亡的影响。ELISA检测1 g/L、0.5 g/L、0.25 g/L药物对LPS处理小鼠单核巨噬细胞白血病细胞RAW264.7后IL-10、TNF-α表达的影响。结果①0.5 g/L板蓝根活性部位作用24 h对Hep-2、Hela、Jukart、U937细胞凋亡无明显影响,但HSV-1感染U937后药物抗病毒组较病毒组的细胞凋亡率由9.72%下降至6.71%,差异有统计学意义(P<0.01);药物作用于Hela细胞时引起其G1期阻滞(从68.76%延长至74.77%),S期缩短(从31.22%降至21.62%),与正常Hela细胞组比较,差异有统计学意义(P<0.05)。②LPS刺激RAW264.7后,与细胞对照组比较,LPS模型组细胞凋亡率升高,差异有统计学意义(P<0.01)。LPS刺激1 g/L、0.5 g/L、0.25 g/L药物组细胞凋亡率分别为19.35%、28.14%、7.52%,明显低于LPS模型组(70.42%),差异有统计学意义(P<0.01)。与细胞对照组比较,1 g/L药物对照组细胞凋亡率上升(由5.81%升至15.03%),差异有统计学意义(P<0.01),而0.5 g/L、0.25 g/L药物对照组无明显促进细胞凋亡的效应。与LPS刺激0.5 g/L、0.25 g/L药物组比较,0.5 g/L、0.25 g/L药物对照组的细胞凋亡率明显下降,差异有统计学意义(P<0.01);与细胞对照组比较,1 g/L药物对照组RAW264.7G1、S期缩短,G2期延长,差异有统计学意义(P<0.01);0.5 g/L药物对照组细胞G1期延长,S期缩短,差异有统计学意义(P<0.01)。③LPS刺激RAW264.7后,与细胞对照组比较,LPS模型组IL-10、TNF-α明显升高,差异有统计学意义(P<0.05);0.5 g/L、0.25 g/L药物对照组IL-10降低,1 g/L药物对照组TNF-α升高,差异有统计学意义(P<0.05)。与LPS模型组比较,LPS 1 g/L、0.5 g/L、0.25 g/L IL-10刺激药物组降低,差异有统计学意义(P<0.05)。结论板蓝根活性部位抑制HSV-1和LPS引起U937、RAW264.7的细胞凋亡,并24 h时下调LPS作用后RAW264.7的IL-10表达。
Objective To investigate the effects of the active site of Banlangen on the apoptosis of Hep-2, Hela, U937 and Jukart cells and the expression of cytokines IL-10 and TNF-α of RAW264.7 cells in mouse monocyte-macrophage leukemia cells. Methods Hep-2, Hela, U937 and Jukart were treated with 0.5 g / L of Radix Isatidis for 24 h. The effect of HSV-1 on the apoptosis was investigated by PI staining and flow cytometry. U937, 0.5 g / L drug treatment 24 h, flow cytometry to study the impact of apoptosis. The effect of 1 g / L, 0.5 g / L and 0.25 g / L of drugs on the expression of IL-10 and TNF-α in RAW264.7 cells induced by LPS in mouse mononuclear macrophage leukemia cells was detected by ELISA. Results ① The active site of 0.5 g / L Banlangen had no significant effect on the apoptosis of Hep-2, Hela, Jukart and U937 cells after 24 h treatment, but the apoptosis rate of Hep-2, Hela, Jukart and U937 cells after HSV- (P <0.01). The G1 phase arrest (from 68.76% to 74.77%) and S phase shortening (from 31.22% to 21.62 %), Compared with the normal Hela cell group, the difference was statistically significant (P <0.05). (2) After LPS stimulated RAW264.7 cells, compared with the cell control group, the apoptosis rate of LPS model group increased, the difference was statistically significant (P <0.01). The apoptotic rate of LPS-treated group was 19.35%, 28.14% and 7.52%, respectively, which was significantly lower than that of LPS group (70.42%) at 1 g / L, 0.5 g / L and 0.25 g / P <0.01). Compared with the cell control group, the apoptosis rate of 1 g / L drug control group increased from 5.81% to 15.03%, the difference was statistically significant (P <0.01), while 0.5 g / L, 0.25 g / L drug The control group did not significantly promote apoptosis. Compared with the drug group of 0.5 g / L and 0.25 g / L LPS, the apoptotic rates of 0.5 g / L, 0.25 g / L drug control group were significantly decreased, the difference was statistically significant (P <0.01); compared with the cell control Compared with the control group, the cell cycle of RAW264.7G1 and S in 1 g / L group was shortened and the G2 phase was prolonged (P <0.01). The G1 phase and the S phase of 0.5 g / L drug control group were shortened, There was statistical significance (P <0.01). (3) Compared with the cell control group, the levels of IL-10 and TNF-α in LPS model group were significantly increased after RAW 264.7 LPS stimulation (P <0.05); compared with the control group IL-10 decreased, 1g / L drug control group increased TNF-α, the difference was statistically significant (P <0.05). Compared with LPS model group, LPS 1 g / L, 0.5 g / L, 0.25 g / L IL-10 stimulated drug group decreased, the difference was statistically significant (P <0.05). Conclusion The active site of Banlangen could inhibit the apoptosis of U937 and RAW264.7 cells induced by HSV-1 and LPS, and the expression of IL-10 in RAW264.7 cells induced by LPS at 24 h.