柚皮素对K562细胞的增殖抑制作用及其作用机制研究

Effect and expression of HO-1 and Bcl-2 in naringenin-treatd K562 cell

目的 观察柚皮素（naringenin,NG）对人慢性髓系白血病K562细胞株的增殖抑制作用及其对HO-1蛋白和Bcl-2蛋白表达的影响。方法 将不同浓度的NG分别作用于K562细胞12、24、48小时,采用MTT法检测NG对K562细胞的生长抑制作用;采用流式细胞仪检测NG对K562细胞的诱导凋亡作用;免疫组织化学S-P法观察NG在不同浓度下作用于K562细胞48小时后,HO-1蛋白和Bcl-2蛋白的表达变化,采用光密度分析其结果,并对二者进行相关性分析。结果 NG 0、100、200、400、800μmol/L体外培养K562细胞12、24、48小时,NG 100-400μmol/L对K562细胞生长呈剂量依赖性抑制（P〈0.05）;在同一浓度下,NG对K562细胞生长呈时间依赖性抑制（P〈0.05）;同一浓度NG作用下,NG诱导的K562细胞凋亡率随培养时间的延长而升高（P〈0.05）,同一培养时间NG诱导的K562细胞凋亡率随NG浓度的增加而升高（P〈0.05）,提示NG诱导K562细胞凋亡的作用呈剂量依赖性增强;K562细胞与不同浓度NG培养48小时,随着NG浓度的升高,K562细胞的HO-1蛋白和Bcl-2蛋白表达逐渐下降（r=-0.884,r=-0.864,P〈0.05）,HO-1蛋白表达与Bcl-2蛋白表达呈正相关（r=0.950,P〈0.05）。结论 NG对K562细胞具有明显的增殖抑制和诱导凋亡作用,其机制可能是通过减少K562细胞HO-1蛋白和Bcl-2蛋白的表达来实现。

Objective To investigate the effect and expression of HO-1 protein and Bcl-2 protein in naringenin-treated K562 cells, and to study the possible mechanism of apoptosis induced by naringenin. Method K562 cells were cultured in different concentration ofnaringenin for 12, 24 and 48 hours. MTT and flow cytometry were used to assess cell growth inhibition and apoptosis of K562 cells after treatment with naringenin respectively. In addition, the expression of HO-1 protein and Bcl-2 proetin in K562 cells in the presence of naringenin for 48 hours was detected by immunohistochemical S-P method. Data were analyzed using Image-pro plus software and correlation analysis. Result K562 cells were cultured in the presence of naringenin （0, 100, 200, 400 and 800 panol/L） for 12, 24 and 48 hours and analyzed by MTT. The growth inhibition rate of naringenin-treated K562 increased in a dose denpendent manner as the concentration of naringenin growed from 0 μmol/L to 400 μmol/L （P 〈 0.05）. The growth inhibition rate of naringenin-treated K562 increased in a time denpendent manner （P 〈 0.05）; K562 cells cultured in 0, 100, 200, 400 and 800 μmol/L naringenin for 12, 24 and 48 hours were detected by FCM. The apoptosis rate of naringenin-treated K562 cells increased in a dose and time dependent manner （P 〈 0.05）. The expression of HO-1 protein and Bcl-2 protein in naringenin-treated K562 cells which measured by immunohistochemical S-P method decreased as the concentration increased; correlation analysis were assayed among naringenin, the expression of HO-1 and Bcl-2. There was a significant negative correlation between concentration of naringenin and expression level of HO-1 and Bcl-2 （r = -0.884, r = -0.864, P 〈 0.05）, and a significant correlation between the expression of HO-1 and Bcl-2 （r = 0.950, P 〈 0.05）. Conclusion Naringenin has a significant growth inhibitory effect and apoptotic effect on K562 ceils, which may be conducted by down-regulation of riO-1 and Bcl-2.