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目的毛萼乙素(Eriocalyxin B,EriB)是一种从唇形科植物疏花毛萼香茶菜中提取的二萜类化合物,具有很强的抗肿瘤细胞活性。本研究探讨EriB对非霍奇金淋巴瘤(non-Hodgkin’s lymphoma,NHL)细胞增殖抑制及凋亡诱导、周期阻滞作用及机制。方法采用MTT法检测5个浓度(0、0.2、0.4、0.6、0.8和1μmol/L)EriB处理72h淋巴瘤细胞株的增殖抑制作用;流式细胞仪检测细胞凋亡率和细胞周期变化;蛋白质印迹法分析不同信号通路蛋白的表达。结果 0、0.2、0.4、0.6、0.8和1μmol/L浓度的EriB对淋巴瘤细胞均有增殖抑制作用,且抑制作用随着浓度增加而增强,半数生长抑制浓度在0.2~1.0μmol/L。流式细胞术检测显示,0.4μmol/L EriB作用Namalwa细胞24h后G0/G1期细胞百分率为(44.71±4.10)%,对照组为(33.79±1.86)%,差异有统计学意义,Z=17.78,P=0.041;FCM结果显示,分析0.4μmol/L EriB作用Namalwa细胞6、12和24h凋亡率分别为(4.21±1.06)%、(20.34±4.71)%和(29.91±5.39)%,显著高于对照组的(3.04±0.96)%,差异有统计学意义,H=8.56,P<0.001。蛋白质印迹法分析显示,EriB处理后的Namalwa细胞蛋白p21、p27、p-Caspase-3、p-Caspase-8和p-Caspase-9表达上调,磷酸化NF-κB和其激活子IκB激酶表达下调。结论 EriB可能通过下调NF-κB信号通路诱导细胞凋亡,使细胞周期阻滞于G0/G1期,抑制淋巴瘤细胞增殖。
Objective Eriocalyxin B (EriB) is a diterpenoid extracted from the genus Labiatae of the genus Labiatae and has strong anti-tumor activity. This study was aimed to investigate the effects of EriB on cell proliferation, apoptosis induction and cell cycle arrest in non-Hodgkin’s lymphoma (NHL) and its mechanism. Methods MTT assay was used to detect the inhibitory effects of 5 concentrations (0, 0.2, 0.4, 0.6, 0.8 and 1 μmol / L) of EriB on 72h lymphoma cell lines. Flow cytometry was used to detect apoptosis and cell cycle changes. Proteins Western blot analysis of different signaling pathway protein expression. Results EriB at concentrations of 0, 0.2, 0.4, 0.6, 0.8 and 1 μmol / L had inhibitory effects on the proliferation of lymphoma cells. The inhibitory effect was enhanced with the increase of concentration. The inhibitory concentration of 50% was 0.2-1.0 μmol / L. Flow Cytometry showed that the percentage of cells in G0 / G1 phase of Namalwa cells treated with 0.4μmol / L EriB was (44.71 ± 4.10)% and that of control group was (33.79 ± 1.86)%, the difference was statistically significant (Z = 17.78 , P = 0.041. The results of FCM showed that the apoptosis rates of Namalwa cells at 4μmol / L EriB were (4.21 ± 1.06)%, (20.34 ± 4.71)% and (29.91 ± 5.39)% Higher than the control group (3.04 ± 0.96)%, the difference was statistically significant, H = 8.56, P <0.001. Western blot analysis showed that the expressions of p21, p27, p-Caspase-3, p-Caspase-8 and p-Caspase-9 were up-regulated in EriB-treated Namalwa cells. The phosphorylation of NF-κB and its activator of IκB kinase were down-regulated . Conclusion EriB may induce cell apoptosis by down-regulating NF-κB signaling pathway, arresting cell cycle in G0 / G1 phase and inhibiting lymphoma cell proliferation.