红花是一种常用的活血化瘀药,羟基红花黄色素A(HSYA)是其主要的活性成分。为了研究HSYA对脂多糖(LPS)诱导的白细胞(PMN)损伤的保护作用,采用比色法来观察HSYA对LPS诱导PMN的黏附作用、Fura-2法测定PMN内Ca2+浓度、RT-PCR法测定TNF-α、IL-6 mRNA表达水平及免疫荧光染色法观察NF-κB核移位的情况。实验结果显示:HSYA浓度为1.01×10-4 mol·L-1时就可明显缓解LPS诱导的家兔PMN黏附活性增强;浓度为3.1×10-5 mol·L-1时可抑制细胞内游离Ca2+浓度升高;浓度为5.2×10-5 mol·L-1时可缓解TNF-α、IL-6 mRNA表达水平的升高,并可抑制NF-κB p65亚基的核移位。结果说明,HSYA对LPS诱导的PMN活化具有缓解作用。
Carthamus tinctorius L.is a traditional Chinese medicine with the effect of promoting blood circulation and removing blood stasis.HSYA(hydroxysafflor yellow A) is the main effective component of Carthamus tinctorius L.In order to study the inhibitory effects of HSYA against PMN(polymorphonuclear) activation induced by LPS(lipopolysaccharide),rabbit PMN adhesion potency which was activated by LPS through colorimetry method was observed.Cellular free calcium concentration was determined by fluorescence spectrophotometry.RT-PCR was applied to study the effect of HSYA on PMN TNF-α and IL-6 mRNA expression;The inhibition of HSYA on NF-κB activation was monitored with immunofluorescence.The results showed that after treated with HSYA,the increase of adhesion potency(HSYA dose 1.01×10-4 mol·L-1),free calcium concentration(HSYA dose 3.1×10-5 mol·L-1),TNF-α and IL-6 mRNA expression elevation(HSYA dose 5.2×10-5 mol·L-1) induced by LPS were inhibited.HSYA can inhibit NF-κB p65 subgroup nuclear translocation(HSYA dose 5.2×10-5 mol·L-1).It is suggested that HSYA is effective in PMN activation induced by LPS.