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） 5 μ g/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) 5 μg/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⁺, pNF- κB⁺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.

R962.2