Houng Jer-Yiing, Tai Tzong-Shyuan, Hsu Shu-Ching, Hsu Hsia-Fen, Hwang Tzann-Shun, Lin Chih-Jiun, Fang Li-Wen
Department of Nutrition, I-Shou University, No.8, Yida Rd., Yanchao District, Kaohsiung City, 82445 Taiwan.
Department of Medical Research, E-Da Hospital, Kaohsiung City, 82445 Taiwan.
Chin Med. 2017 Apr 11;12:9. doi: 10.1186/s13020-017-0130-4. eCollection 2017.
(GT) () is a Chinese herbal medicine previously exhibited an anti-inflammatory activity. This study aimed to investigate the effect of GT ethanol extract (GTE) on T cell-mediated adaptive immunity.
Human peripheral blood mononuclear cells (PBMCs) and Jurkat T cells were activated by phytohemagglutinin in the presence of various doses (3.13-50 μg/mL) of GTE. The effect of GTE on T cell activation was examined by a proliferation assay of activated PBMCs and the level of the activation marker CD69 on the surface of activated Jurkat T cells. Apoptosis was determined by propidium iodide staining in hypotonic solution. Signaling pathway molecules were assessed by western blotting.
ethanol extract was demonstrated to inhibit T cell activation, not only in the proliferation of human PBMCs at the concentrations of 12.5, 25 and 50 μg/mL ( = 0.0118, 0.0030 and 0.0021) but also in the CD69 expression in Jurkat cells, which was not due to the cytotoxicity of GTE. The presence of GTE did not change the activity of nuclear factor kappa-light-chain-enhancer of activated B cells or extracellular signal-regulated kinase upon T cell activation. In addition, GTE significantly reduced activation of c-Jun N-terminal kinase (JNK) ( = 0.0167) and p38 ( = 0.0278). Furthermore, decreased JNK activation mediated the preventive effect of GTE on T cell activation-induced cell death (AICD).
ethanol extract inhibited T cell activation of Jurkat cells and freshly prepared human PBMCs due to suppression of JNK activity. Furthermore, GTE inhibited AICD by blocking prolonged JNK phosphorylation in activated T cells. Taken together, the anti-inflammatory effects exerted by GTE were mediated via suppression of JNK phosphorylation in T cell activation.
（GT）（）是一种先前已显示出抗炎活性的中草药。本研究旨在探讨GT乙醇提取物（GTE）对T细胞介导的适应性免疫的影响。
在存在不同剂量（3.13 - 50μg/mL）GTE的情况下，用植物血凝素激活人外周血单核细胞（PBMC）和Jurkat T细胞。通过活化PBMC的增殖试验以及活化Jurkat T细胞表面活化标志物CD69的水平来检测GTE对T细胞活化的影响。通过低渗溶液中的碘化丙啶染色来测定细胞凋亡。通过蛋白质印迹法评估信号通路分子。
乙醇提取物被证明可抑制T细胞活化，不仅在浓度为12.5、25和50μg/mL时抑制人PBMC的增殖（P = 0.0118、0.0030和0.0021），还抑制Jurkat细胞中CD69的表达，这并非由于GTE的细胞毒性所致。GTE的存在在T细胞活化时并未改变活化B细胞核因子κ轻链增强子或细胞外信号调节激酶的活性。此外，GTE显著降低了c - Jun氨基末端激酶（JNK）的活化（P = 0.0167）和p38的活化（P = 0.0278）。此外，JNK活化的降低介导了GTE对T细胞活化诱导的细胞死亡（AICD）的预防作用。
乙醇提取物由于抑制JNK活性而抑制Jurkat细胞和新鲜制备的人PBMC的T细胞活化。此外，GTE通过阻断活化T细胞中JNK的长时间磷酸化来抑制AICD。综上所述，GTE发挥的抗炎作用是通过抑制T细胞活化中的JNK磷酸化介导的。
