补骨脂乙醇提取物对人T细胞亚群活化增殖及AKT/NF-κB信号通路的调节作用

Effects of ethanol extract from Psoralea corylifolia on activation and proliferation of human T cell subsets and regulation of Akt/NF-κB signaling pathway

目的探讨补骨脂乙醇提取物(PCEE)对人T细胞亚群增殖和活化的影响。方法采集健康人EDTA抗凝血,分离外周血单个核细胞(PBMCs),ADD染色流式细胞术检测PCEE(1、2 mg/mL)对PBMCs的毒性;PBMCs分为对照组、模型组、PCEE(1 mg/mL)组,模型组和PCEE组用植物血凝素(PHA)5μg/mL刺激,对照组不加;3 d后PCEE组加入终质量浓度为1 mg/mL的PCEE作用48 h,对照组和模型加入等体积的PBS。48 h后收集细胞,用流式细胞术检测CD4^(+)和CD8^(+)T细胞亚群增殖(ADD染色)、活化(CD69和CD25)以及AKT、NF-κB磷酸化水平。结果PCEE 1 mg/mL组活细胞比例为(93.06±2.12)%,与对照组(95.16±1.39)%比较无显著差别;PCEE组CD4^(+)和CD8^(+)T细胞的增殖显著低于模型组(P<0.001);PCEE组CD69^(+)CD4^(+)、CD69^(+)CD8^(+)和CD25^(+)CD4^(+)、CD25^(+)CD8^(+)T细胞比例显著低于模型组(P<0.05);PCEE组pAKT^(+)CD4^(+)、pAKT^(+)CD8^(+)和pNF-κB^(+)CD4^(+)、pNF-κB^(+)CD8^(+)T细胞比例显著低于模型组(P<0.05)。结论补骨脂通过抑制AKT/NF-κB信号传导通路显著抑制T细胞增殖和活化。

Objective To investigate the effects of ethanol extract from Psoralea corylifolia(PCEE)on proliferation and activation of T cell subsets.Methods EDTA anticoagulant blood was collected from healthy subjects,and peripheral blood mononuclear cells(PBMCs)were isolated,ADD staining flow cytometry was used to detect the toxicity of PCEE(1,2 mg/mL)to PBMCs.PBMCs were divided into control group,model group,and PCEE(1 mg/mL)group.PBMCs in model group and PCEE(1 mg/mL)group were stimulated with phytohemagglutinin(PHA)5μg/mL for 3 d,the control group was not added.PCEE group was added with a final mass concentration of 1 mg/mL for 48 h,control group and model were added with equal volume of PBS.Cells were collected 48 h later,and the proliferation(ADD staining),activation(CD69 and CD25),AKT and NF-κB phosphorylation levels of CD4^(+)and CD8^(+)T cell subsets were detected by flow cytometry.Results The proportion of living cells in PCEE 1 mg/mL group was(93.06±2.12)%,which was not significantly different from that in control group(95.16±1.39)%.The proliferation of CD4^(+)and CD8^(+)T cells in PCEE group was significantly lower than that in model group(P<0.001).The proportion of CD69^(+)CD4^(+),CD69^(+)CD8^(+)and CD25^(+)CD4^(+),CD25^(+)CD8^(+)T cells in PCEE group was significantly lower than that in model group(P<0.05).The proportion of pAKT^(+)CD4^(+),pAKT^(+)CD8^(+)and pNF-κB^(+)CD4^(+),pNFκB^(+)CD8^(+)T cells in PCEE group was significantly lower than that in model group(P<0.05).Conclusion This study showed that PECC significantly inhibited T cell proliferation and activation by inhibiting theAkt/NF-κB signaling pathway.