目的 探討補骨脂乙醇提取物（PCEE）對人T細胞亞群增殖和活化的影響。方法 采集健康人EDTA抗凝血，分離外周血單個核細胞（PBMCs），ADD染色流式細胞術(shù)檢測PCEE（1、2 mg/mL）對PBMCs的毒性；PBMCs分為對照組、模型組、PCEE （1 mg/mL）組，模型組和PCEE組用植物血凝素（PHA） 5 μ g/mL刺激，對照組不加； 3 d后PCEE組加入終質(zhì)量濃度為1 mg/mL的PCEE作用48 h，對照組和模型加入等體積的PBS。48 h后收集細胞，用流式細胞術(shù)檢測CD4⁺和CD8⁺T細胞亞群增殖（ADD染色）、活化（CD69和CD25）以及AKT、NF-κB磷酸化水平。結(jié)果 PCEE 1 mg/mL組活細胞比例為（93.06±2.12）%，與對照組（95.16±1.39）%比較無顯著差別；PCEE組CD4⁺和CD8⁺T細胞的增殖顯著低于模型組（P<0.001）；PCEE組CD69⁺CD4⁺、CD69⁺CD8⁺和CD25⁺CD4⁺、CD25⁺CD8⁺T細胞比例顯著低于模型組（P<0.05）；PCEE組pAKT⁺CD4⁺、pAKT⁺CD8⁺和pNF-κB⁺CD4⁺、pNF-κB⁺CD8⁺T細胞比例顯著低于模型組（P<0.05）。結(jié)論 補骨脂通過抑制AKT/NF-κB信號傳導(dǎo)通路顯著抑制T細胞增殖和活化。

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