Department of Dermatology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Sugitani, Toyama, Japan.
Inflamm Res. 2012 Nov;61(11):1177-85. doi: 10.1007/s00011-012-0512-0. Epub 2012 Jul 1.
Platinum nanoparticles (nano-Pt) have been reported to possess anti-oxidant and anti-tumor activities. However, the biological activity and mechanism of action of nano-Pt in inflammation are still unknown. The present study was designed to determine the in-vitro anti-inflammatory effects of nano-Pt on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells.
RAW 264.7 macrophages were used for the study. The LPS-induced production of reactive oxygen species (ROS) was determined by flow cytometry. The prostaglandin E(2) (PGE(2)) concentration was measured using a PGE(2) assay kit. The protein levels and mRNA expression of the pro-inflammatory cytokines [tumor necrosis factor-α, interleukin (IL)-1β and IL-6], along with cyclooxygenase (COX-2) and inducible nitric oxide synthase, were analyzed by Western blotting and reverse transcription-polymerase chain reaction analysis. The phosphorylation of extracellular signal regulated kinase (ERK1/2) and Akt, and the phosphorylation and degradation of inhibitory kappa B-alpha (IκB-α) was determined by Western blot analysis.
Nano-Pt significantly reduced the LPS-induced production of intracellular ROS and inflammatory mediators. In addition, nano-Pt suppressed the phosphorylation of ERK1/2 and Akt, and significantly inhibited the phosphorylation/degradation of IκB-α as well as nuclear factor kappa-B (NFκB) transcriptional activity.
These results suggest that the anti-inflammatory properties of nano-Pt may be attributed to their downregulation of the NFκB signaling pathway in macrophages, thus supporting the use of nano-Pt as an anti-inflammatory agent.
铂纳米粒子(nano-Pt)已被报道具有抗氧化和抗肿瘤活性。然而,nano-Pt 在炎症中的生物学活性和作用机制尚不清楚。本研究旨在确定 nano-Pt 对脂多糖(LPS)刺激的 RAW 264.7 细胞的体外抗炎作用。
使用 RAW 264.7 巨噬细胞进行研究。通过流式细胞术测定 LPS 诱导的活性氧(ROS)的产生。使用 PGE2 测定试剂盒测定前列腺素 E2(PGE2)浓度。通过 Western blot 和逆转录聚合酶链反应分析,分析促炎细胞因子[肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和 IL-6]、环氧合酶(COX-2)和诱导型一氧化氮合酶的蛋白水平和 mRNA 表达。通过 Western blot 分析测定细胞外信号调节激酶(ERK1/2)和 Akt 的磷酸化,以及抑制性κB-α(IκB-α)的磷酸化和降解。
nano-Pt 显著降低了 LPS 诱导的细胞内 ROS 和炎症介质的产生。此外,nano-Pt 抑制了 ERK1/2 和 Akt 的磷酸化,并显著抑制了 IκB-α的磷酸化/降解以及核因子 kappa-B(NFκB)转录活性。
这些结果表明,nano-Pt 的抗炎特性可能归因于其对巨噬细胞中 NFκB 信号通路的下调,从而支持将 nano-Pt 用作抗炎剂。