紫草素抑制人气道平滑肌细胞的增殖

目的 研究紫草素对体外培养的人气道平滑肌细胞(HASMCs)增殖的影响.方法 组织块贴壁法原代培养传至4～8代的HASMCs,经40、20、10、5、2、1、0.5 ?mol/L及0 ?mol/L(对照组)的紫草素分别作用12、24、48 h后,应用MTS法测定紫草素对HASMCs的细胞增殖的影响;经40、20、10、5 ?mol/L及0 ?mol／L(对照组)的紫草素分别作用24 h后,应用流式细胞术分析紫草素对细胞周期的影响;经20 ?mol/L及0 ?mol/L(对照组)的紫草素分别作用24 h后,应用SP免疫细胞化学法检测紫草素对增殖细胞核抗原(PCNA)表达的影响.结果 (1)与对照组相比,20 ?mol/L和40 ?mol/L的紫草素均可有效抑制细胞增殖(均P<0.05),以48 h最为显著(抑制率分别为30.1％与42.9％),但两者之间无显著性差异(P0.05).(2)20 ?mol/L及40 ?mol/L的紫草素均可显著提高G0/G1期比例(P<0.05),阻滞细胞周期进程,但两者之间无显著性差异(P0.05).(3)20 ?mol/L的紫草素可显著下调PCNA的表达(P<0.01).结论 紫草素对HASMCs具有明确的抗增殖作用.

Shikonin inhibits the proliferation of human airway smooth muscle cells

OBJECTIVE: To study the inhibitory effect of shikonin on the proliferation of human airway smooth muscle cells (HASMCs) in vitro. METHODS: HASMCs from the trachea were obtained by primary culture of the tissue explants and adherent culture. The HASMCs were exposed to shikonin at 0 (control group), 0.5, 1, 2, 5, 10, 20, and 40 micromol/L for 12, 24, and 48 h, after which the cell proliferation was assessed by 3-(4,5-carboxymethoxypheny1)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay. Flow cytometry was used for cell cycle analysis of the HASMCs exposed to shikonin at 40, 20, 10, 5 micromol/L and 0 micromol/L (control group) for 24 h. Immunocytochemistry with SP method was performed to detect the expression of proliferating cell nuclear antigen (PCNA) in the HASMCs treated with shikonin at 20 micromol/L and 0 micromol/L (control group) for 24 h. RESULTS: Shikonin at the concentrations of 20 and 40 micromol/L significantly inhibited the proliferation of the cells (P<0.05), and the effect was especially obvious after 48 h of cell exposure, with inhibition rates of 30.1% and 42.9%, respectively. No significant difference was found between the two concentrations for their cell growth inhibition effects (P>0.05). Shikonin at the concentrations of 20 and 40 micromol/L caused significant cell cycle arrest in G(0)/G(1) phase (P<0.05), the effect of which, however, was not concentration-dependent (P>0.05). Shikohin at 20 micromol/L significantly down-regulated the expression of PCNA in the cells (P<0.05). CONCLUSION: Shikonin can inhibit the proliferation of HASMCs in vitro.