华中枸骨黄烷类成分体外抗炎与自由基清除活性

In vitro anti-inflammatory and free radical scavenging activities of flavans from Ilex centrochinensis

为研究华中枸骨中多个黄烷类成分（1-6）体外抗炎与自由基清除活性，并初步分析其构效关系。采用脂多糖诱导小鼠RAW264．7巨噬细胞炎症模型，MTT法测定细胞毒性作用，Griess法测定NO生成量，ELISA法检测细胞上清液中TNF—α，IL-1β，IL-6，PGE2的含量；体外测定清除DPPH自由基，超氧阴离子自由基和羟自由基活性。结果表明黄烷1，3，4，6体外显示很强的抗炎活性，能显著抑制NO，TNF-α，IL-1β和IL-6的分泌，化合物1的抑制作用最强，黄烷2和5显示相对较弱的抑制活性，体外自由基清除活性顺序与抗炎活性相似。因此，黄烷1，3，4，6是华中枸骨抗炎和自由基清除活性的活性成分，构效关系初步分析表明4’一羟基为黄烷类化合物显示抗炎和自由基清除活性的主要结构部位。

This study was carried out to evaluate the anti-inflammatory and free radical scavenging activities of flavans from [lex cen- trochinensis S.Y. Hu in vitro and their structure-activity relationship. LPS-stimulated RAW 264. 7 macrophage was used as inflammato- ry model. MTT assay for cell availability, Griess reaction for nitric oxide （NO） production, the content of TNF-α, IL-1β, IL-6 and PGE2 were detected with ELISA kits; DPPH, superoxide anion and hydroxyl free radicals scavenging activities were also investigated. According to the result, all flavans tested exhibited anti-inflammatory effect in different levels. Among them, compounds 1, 3, 4 and 6 showed potent anti-inflammatory effect through the inhibition of NO, TNF-α, IL-1β and IL-6, of which 1 was the most effective inhibi- tor, however, 2 and 5 were relatively weak or inactive. The order of free radical scavenging activities was similar to that of anti-inflam- matory activities. Therefore, these results suggest that 3, 4 and 6, especially of 1, were,in part responsible for the anti-inflammatory and free radical scavenging activity of llex centrochinensis. Hydroxyl group at 4＇-position of B-ring plays an important role in the anti-in- flammatory and free radical scavenging capacities.