目的 研究苦參堿對(duì)原代人白血病細(xì)胞的體外作用及可能的分子機(jī)制。方法 取臨床初診的髓系白血病患者外周血, 分離得到原代白血病細(xì)胞體外培養(yǎng)。不同濃度苦參堿作用后, 觀察細(xì)胞的形態(tài)學(xué)改變、CCK8法檢測(cè)細(xì)胞增殖、Annexin V-FITC/PI分析細(xì)胞凋亡及流式檢測(cè)細(xì)胞周期。結(jié)果 苦參堿作用24 h, 細(xì)胞增殖明顯受抑, 0.5、0.8 mg/mL組的生長(zhǎng)抑制率分別為54.98%和72.96%, 與對(duì)照組相比差異顯著(P<0.01), 抑制效應(yīng)有明顯劑量相關(guān)性?？鄥A作用24 h的IC₅₀為0.5 mg/mL?？鄥A作用24 h, 0.2、0.5、0.8 mg/mL組細(xì)胞早期凋亡率分別為11.8%、37.6%、54.7%, 明顯高于對(duì)照組自發(fā)凋亡率(7.50%)(P<0.05)?？鄥A作用48 h后, 細(xì)胞周期阻滯于G0/G1→S期。結(jié)論 苦參堿可顯著抑制體外培養(yǎng)的人原代白血病細(xì)胞增殖, 其機(jī)制與誘導(dǎo)細(xì)胞早期凋亡, 促進(jìn)細(xì)胞周期G1→S期阻滯有關(guān)。

Objective To investigate the effects of matrine on the human primary 1eukemia cells and its possible molecular mechanisms. Methods The primary leukemia cells were isolated from the PBMC of the patients with myeloid leukemia and cultivated under the regular culture medium. After exposed to the matrine with different concentration, the morphological changes of leukemia cells were observed under the light biomicroscopy. The proliferation was determined by CCK-8 assay. Annexin V-FITC/PI affinity assay was used to analyze the apoptosis of leukemia cells induced by matrine. The cell cycles were analyzed by flow cytometry before and after matrine treatment. Results Compared to the vehicle cells, there was a dramatically proliferation suppression was observed in the cells after matrine treatment. The inhibitory rates of primary leukemia cells were 54.98% and 72.96% after treated with matrine for 24 h at the doses of 0.5 and 0.8 mg/mL, respectively (P < 0.01), which has a significant dose-dependent manner. The half maximal inhibitory concentration of matrine (IC₅₀) was 0.5 mg/mL for 24 h treatment. Matrine could induce early cell apoptosis. The percentage of apoptotic cells were 11.8%, 37.6%, and 54.7% in the cells treated with 0.2, 0.5, or 0.8 mg/mL matrine for 24 h, respectively (with spontaneous apoptosis rate 7.50% for untreated cells) (P < 0.05). FCM method showed that cells at the G0/G1 phase decreased and cells at the S phase were declined obviously, suggested a blockage of cell cycles transition at the G1/S key checkpoint for 48 h. Conclusion Matrine manifests a significant inhibitory effects on the proliferation of human primary leukemia cells. Apoptosis induction and the arrest at G1 phase of cell cycle maybe contribute to its roles on the suppression of cell growth.

國家自然科學(xué)基金(81101647);南京醫(yī)科大學(xué)博士后課題