Shen Yuh-Chiang, Wang Yea-Hwey, Chou Yueh-Ching, Chen Chieh-Fu, Lin Lie-Chwen, Chang Tun-Tschu, Tien Jung-Hsiung, Chou Cheng-Jen
National Research Institute of Chinese Medicine, Taipei, Taiwan.
Planta Med. 2004 Apr;70(4):310-4. doi: 10.1055/s-2004-818941.
We have previously shown that a concentrated ethanol extract of the fruiting bodies of Antrodia camphorata exhibited immunomodulating effects in human leukocytes and fourteen compounds including zhankuic acids A, B, C, and antcin K were identified in the extract. In this study, an acute cellular model in isolated peripheral human neutrophils was established to elucidate the anti-inflammatory effects of these compounds. Reactive oxygen species (ROS) production and firm adhesion by neutrophils display two important responses during inflammation. To evaluate whether these compounds could prevent inflammatory responses by neutrophils, their effects on N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate-13-acetate (PMA)-activated peripheral human neutrophils were examined. Pretreatment with 1 - 25 microM of zhankuic acids A, B, C, or antcin K concentration-dependently diminished fMLP- or PMA-induced ROS production, as measured by a lucigenin-amplified chemiluminescence, with IC (50) (microM) around 5 - 20 microM. Zhankuic acids A, B, C, or antcin K also effectively inhibited the fMLP- or PMA-induced firm adhesion without interfering with the up-expression of surface Mac-1 (CD11b/CD18), a beta2 integrin mediating the firm adhesion of neutrophils to endothelium. The anti-inflammatory actions of these drugs were not due to cytotoxic effects because no significant difference in cell viability was observed compared to vehicle control. These data suggest that inhibition of both ROS production and firm adhesion by neutrophils has no significant cytotoxic effect that could give these drugs the potential to be anti-inflammatory agents for the clinical treatment.
我们之前已经表明，樟芝子实体的浓缩乙醇提取物在人白细胞中表现出免疫调节作用，并且在该提取物中鉴定出了包括展青霉素酸A、B、C和樟芝酸K在内的14种化合物。在本研究中，建立了分离的人外周血中性粒细胞急性细胞模型，以阐明这些化合物的抗炎作用。中性粒细胞产生活性氧（ROS）和牢固黏附是炎症过程中的两个重要反应。为了评估这些化合物是否能预防中性粒细胞的炎症反应，研究了它们对N-甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）或佛波酯12-肉豆蔻酸酯-13-乙酸酯（PMA）激活的人外周血中性粒细胞的影响。用1 - 25微摩尔的展青霉素酸A、B、C或樟芝酸K预处理，以腔肠素增强的化学发光法测定，可浓度依赖性地减少fMLP或PMA诱导的ROS产生，半数抑制浓度（IC50）约为5 - 20微摩尔。展青霉素酸A、B、C或樟芝酸K也有效抑制fMLP或PMA诱导的牢固黏附，而不干扰表面Mac-1（CD11b/CD18）的上调表达，Mac-1是一种β2整合素，介导中性粒细胞与内皮细胞的牢固黏附。这些药物的抗炎作用并非由于细胞毒性作用，因为与溶剂对照相比，未观察到细胞活力有显著差异。这些数据表明，抑制中性粒细胞产生ROS和牢固黏附没有显著的细胞毒性作用，这使得这些药物有可能成为临床治疗的抗炎剂。
