IMPACT OF A BLACKBERRY EXTRACT AND SINGLE ANTHOCYANINS ON THE INFLAMMATORY RESPONSE OF HUMAN MACROPHAGE THP-1 CELLS

E. Cenk, G. Pahlke, A. Oertel, S. Martens, A. Matros, H. Mock, D. Marko
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Abstract

A diet rich in anthocyanins is considered to be beneficial for health. Thus, investigations on anthocyanin-rich extracts and single constituents are of great interest. We investigated a blackberry extract, containing a spectrum of anthocyanins, in comparison to two sub-fractions comprising cyanidin-3-(6''-dioxalylglucoside and cyanidin-3-(malonylglucoside), respectively, regarding their impact on inflammation-related cytokine expression and secretion in human macrophages. Relative gene transcription and secretion of IL-6 and TNF-alpha were measured using quantitative real time PCR and human cytokine bead-based multiplex immunoassay, respectively. Human THP-1 monocytes were differentiated with phorbol-12-myristate-13-acetate into macrophages which were pre-incubated with physiologically relevant concentrations of blackberry extract or single compounds. Subsequent stimulation with lipopolysaccharides from E.coli induced gene transcription and cytokine release mimicking the inflammatory response. Cyanidin-3-(malonylglucoside) had no impact on LPS-induced IL6 and TNF-alpha expression. In contrast, the blackberry extract and cyanidin-3-(6''-dioxalylglucoside) caused further stimulatory effects for the transcription and secretion of IL-6 at higher concentrations ≥0.05 µg/ml. The secretion of TNF-alpha was increased as well by both, yet with no impact on the transcript level after 3 h of incubation. The results clearly show that single as well as combinatory effects might be of relevance for the immune-stimulatory impact of the blackberry extract.
黑莓提取物和单一花青素对人巨噬细胞thp-1细胞炎症反应的影响
富含花青素的饮食被认为对健康有益。因此,研究富含花青素的提取物和单一成分具有重要意义。我们研究了一种含有花青素光谱的黑莓提取物,并将其与含有花青素-3-(6′-二氧基葡萄糖苷)和花青素-3-(丙二烯基葡萄糖苷)的两个亚组分进行了比较,研究了它们对人类巨噬细胞中炎症相关细胞因子表达和分泌的影响。采用实时荧光定量PCR和基于人细胞因子微球的多重免疫分析法分别检测各组相对基因转录和IL-6、tnf - α的分泌。将人THP-1单核细胞用phorbol12 -肉豆酸酯-13-乙酸酯分化为巨噬细胞,并与生理相关浓度的黑莓提取物或单一化合物预孵育。随后用大肠杆菌脂多糖刺激诱导基因转录和细胞因子释放,模拟炎症反应。花青素-3-(丙二酰葡萄糖苷)对lps诱导的il - 6和tnf - α表达无影响。当浓度≥0.05µg/ml时,黑莓提取物和花青素-3-(6′-二氧基葡萄糖苷)对IL-6的转录和分泌有进一步的刺激作用。培养3小时后,tnf - α的分泌也增加,但对转录物水平没有影响。结果清楚地表明,单一以及组合效应可能与黑莓提取物的免疫刺激作用有关。
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