Yu Wan-ying, Kan Wei-juan, Yu Peng-xia, Li Min-min, Song Ji-shuai, Zhao Feng
School of Pharmacy, Yantai University, Yantai 264005, China.
Zhongguo Zhong Yao Za Zhi. 2012 Sep;37(17):2618-21.
To study and compare the anti-inflammatory effect and molecular mechanism of artemisinin and dihydroartemisinin.
Mouse mononuclear macrophage RAW264.7 cells were stimulated to release inflammatory mediators such as TNF-alpha, IL-6 and NO, in order to assess the drugs' inhibitory effect on macrophage's release of above inflammatory mediators. The levels of TNF-alpha and IL-6 were determined by ELISA and the cytotoxicity was determined by MTT method. The protein expression of iNOS, COX-2 and beta-actin were tested by Western blot. The enzymatic activity of COX-2 was determined by colorimetric method.
Dihydroartemisinin significantly inhibited LPS-induced release of TNF-alpha, IL-6 and NO from RAW264.7 in mice with the concentration range of 12.5 - 100 micromol x L(-1), and showed good dose dependence. Artemisinin only inhibited the IL-6 release to a certain extent.
Dihydroartemisinin inhibits macrophages from releasing inflammatory factors TNF-alpha and IL-6 and inflammatory mediators NO by down-regulating iNOS protein. Artemisinin may help dihydroartemisinin to show its anti-inflammatory effect through metabolism.
研究并比较青蒿素和双氢青蒿素的抗炎作用及分子机制。
刺激小鼠单核巨噬细胞RAW264.7细胞释放肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和一氧化氮（NO）等炎性介质，以评估药物对巨噬细胞释放上述炎性介质的抑制作用。采用酶联免疫吸附测定法（ELISA）测定TNF-α和IL-6水平，采用噻唑蓝（MTT）法测定细胞毒性。通过蛋白质免疫印迹法（Western blot）检测诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）和β-肌动蛋白的蛋白表达。采用比色法测定COX-2的酶活性。
双氢青蒿素在12.5～100 μmol·L⁻¹浓度范围内可显著抑制脂多糖（LPS）诱导的小鼠RAW264.7细胞释放TNF-α、IL-6和NO，并呈现良好的剂量依赖性。青蒿素仅在一定程度上抑制IL-6的释放。
双氢青蒿素通过下调iNOS蛋白抑制巨噬细胞释放炎性因子TNF-α和IL-6以及炎性介质NO。青蒿素可能通过代谢帮助双氢青蒿素发挥抗炎作用。
