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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。
Safflower is a commonly used medicine for promoting blood circulation and removing blood stasis. Hydroxysafflor yellow A (HSYA) is its main active ingredient. To investigate the protective effect of HSYA on lipopolysaccharide (LPS) -induced leukocyte (PMN) injury, the adhesion of HSYA to LPS-induced PMN was observed by colorimetric assay. The Ca2 + concentration in PMN was determined by Fura-2 assay, TNF-α, IL-6 mRNA expression levels and immunofluorescence staining NF-κB nuclear translocation. The experimental results showed that HSYA concentration of 1.01 × 10-4 mol·L-1 could significantly alleviate LPS-induced rabbit PMN adhesion activity enhanced; concentration of 3.1 × 10-5 mol·L-1 can inhibit intracellular free Ca2 + concentration was increased. The concentration of 5.2 × 10-5 mol·L-1 could relieve the increase of TNF-α and IL-6 mRNA expression and inhibit the nuclear translocation of NF-κB p65 subunit. The results showed that HSYA had a relieving effect on LPS-induced PMN activation.