非重型再生障碍性贫血患者血清对32D细胞凋亡及Stat3蛋白磷酸化的影响

Effects of sera from patients with non severe aplastic anemia on the apoptosis and expression of phosphorylated Stat3 protein in 32D cells

目的探讨非重型再生障碍性贫血(NSAA)患者血清对32D细胞凋亡及细胞内信号转导子和转录激活子(Stat3)蛋白磷酸化的影响。方法采集12例NSAA患者(NSAA组)和12例正常健康体检者(对照组)血清,分别孵育小鼠32D细胞后进行细胞计数并检测细胞存活率、凋亡率及细胞内Stat3蛋白的磷酸化水平。结果血清孵育12、24、36、48 h后,NSAA组32D细胞内的Stat3蛋白磷酸化水平均高于对照组,以孵育24 h后差异最大(P<0.05)。孵育24 h后,NSAA组较对照组细胞计数显著减少(P<0.05),细胞存活率低于对照组但无统计学意义,而细胞凋亡率显著高于对照组(P<0.05)。结论 NSAA患者的血清在体外作用24 h后可诱导32D细胞凋亡并上调Stat3蛋白磷酸化水平,Stat3蛋白的磷酸化增多可能参与NSAA患者血清诱导32D细胞凋亡的信号传导过程。

Objective To investigate the expression of phosphorylated Stat3 protein in apoptosis of the myeloid progenitor cell line 32D induced by sera from patients with non severe aplastic anemia（NSAA）.Methods The expressions of phosphorylated Stat3 protein in the 32D cells were determined by Western blot after treated with normal sera or sera from patients with NSAA for 0,12,24,36 and 48 hours.After treated with sera for 24 hours,the 32D cells were subjected to apoptosis analysis using flow cytometry,while cell counting and trypan blue staining were performed to assess cell number and cell viability,respectively.Results The higher expressions of phosphorylated Stat3 protein were found in the NSAA group after treated with sera for 12,24,36 and 48 hours and the difference in the two groups was maximal at 24 hours after treated with sera（P〈0.05）.At 24 hour point after treated with sera,the NSAA group had more apoptotic cells and lower cell numbers compared with those of the control group（P〈0.05）,however,there was no significant difference in cell viability between the two groups.Conclusions 24-hour treatment with sera from patients with NSAA can induce apoptosis of 32D cells and up-regulate the phosphorylated Stat3 expression in vitro.Phosphorylated Stat3 may be involved in the apoptosis of 32D cells induced by sera from patients with NSAA.