花色苷对脂多糖诱导巨噬细胞炎症反应的影响

目的探讨花色苷对脂多糖(LPS)所诱导的THP-1样巨噬细胞中诱导性一氧化氮合酶(iNOS),环氧合酶-2(COX-2)及其产物一氧化氮(NO),前列腺素E2(PGE2)表达的影响。方法THP-1单核细胞用终浓度为167nmol/L的佛波酯(PMA)刺激48h诱导成THP-1样巨噬细胞后,分别用1、10、50、100μmol/L的花色苷标准品矢车菊素-3-葡萄糖苷(Cy-3-g)或芍药素-3-葡萄糖苷(Pn-3-g)孵育细胞2h,再用1μg/mlLPS处理细胞12h,用半定量RT-PCR法分别检测iNOS、COX-2的基因表达量,用Western blotting检测iNOS、COX-2的蛋白表达量,分别用Griess法和碱性磷酸酶法(EIA)检测THP-1样巨噬细胞培养基中NO及PGE2的含量。结果花色苷能有效的下调iNOS,COX-2的基因及蛋白水平,且其所合成的炎性介质NO和PGE2也显著降低。结论花色苷能够降低LPS所诱导的THP-1样巨噬细胞中iNOS,COX-2及其产物NO,PGE2的表达水平,提示其可能对于治疗各种炎症相关性疾病如动脉粥样硬化等有着重要的意义。

EFFECTS OF ANTHOCYANIN ON LIPOPOLYSACCHARIDE-INDUCED INFLAMMATORY RESPONSE IN MACROPHAGES

Objective To explore the effects of anthocyanin on lipopolysaccharide(LPS)-induced nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression with nitric oxide (NO) and prostaglandin E2 (PGE2) release in macrophages. Method THP-1 cells were cultured in the presence of 167 nmol/L phorbol 12-myristate 3-acetate (PMA) for 48 h to induce monocyte–macrophage differentiation. After that,THP-1 macrophages were incubated with 1,10,50 or 100 μmol/L C3G for 2 h and then treated with 1 μg/ml LPS for another 12 h. RT-PCR and Western blotting were used to analyze iNOS and COX-2 gene and protein expression. Additionally,the LPS-induced NO and PGE2 release in macrophages were assayed by the methods of Griess and EIA,respectively. Results Anthocyanins inhibited LPS-induced iNOS and COX-2 expression with NO and PGE2 release in macrophages. Conclusion Anthocyanins can cause inhibition of LPS-induced iNOS and COX-2 at both mRNA and protein levels together with decrease in NO and PGE2 production,indicating that it can be used as a potential anti-inflammatory reagent to treat many inflammatory-related diseases such as atherosclerosis. ACTA NUTRIMENTA SINICA,2009,31(4):366-369]