红花对活化巨噬细胞产生一氧化氮的影响作用

Effects of Carthamus tinctorius L.on nitric oxide release by activated macrophages

目的水提醇沉法对红花进行提取,研究红花提取物对脂多糖(LPS)活化小鼠巨噬细胞RAW264.7产生一氧化氮(NO)的影响作用,同时测定提取物中的总黄酮含量。方法 ATP-Lite法测定红花对RAW264.7细胞毒性的影响,姜黄素为阳性对照。采用Griess法测定由LPS活化RAW264.7细胞产生NO的含量,阿司匹林为阳性对照。以山奈酚为标准品测定红花提取物中总黄酮的含量。结果姜黄素对RAW264.7的半数致死浓度为(29.9±4.3)μg/ml。红花提取物在低于125μg/ml时,对RAW264.7细胞的致死率低于5%。在12.5～100μg/ml的浓度范围内,红花表现出对LPS活化RAW264.7细胞产生NO的抑制作用。以山奈酚计,1g生药红花中含1.14mg总黄酮。结论红花对LPS活化巨噬细胞产生NO具有抑制作用,其作用可能与其所含黄酮类成分有关。

Objective By extraction using aqueous solution and sedimentation using ethanol, we investigated the effects of the major components of Carthamus tinctorius L（C. tinctorius） on nitric oxide （NO） release upon induc-tion with lipopolysaccharide （LPS） in RAW264.7 cells and the level of total fiavonoids in the extracts. Methods Cy-totoxicity on RAW 264.7 cells was evaluated by using the ATP-Lite method in which curcumin served as the positive control. The Griess assay was employed to investigate the level of NO released by RAW264.7 cells, in which aspirin served as the positive control, upon activation by LIPS. The level of total flavonoids was tested based on the standard-ized curve of campherol. Results The half lethal dose （LC50） of curcumin was （29.9±4.3） ;Lg/ml. The extracts yielded a fatality rate of less than 5% within the concentration of 125 μg/ml. An inhibitory effect to NO release by RAW264.7 cells upon LPS-induced activation was evidenced at the concentration of 12.5-100 μg/ml. The content of total flavonoids was 1.14 mg per gram of C.tinctorius by calculation based on that of campherol. Conclusion The fact that C. tinctorius has an inhibitory effect on NO release in activated RAW264.7 macrophage cells might be associated with the pharmacological effects of the flavonoids.