Figure 2
MGDG (monogalactosyldiacylglycerol) anti-inflammatory activity on IL-1α (interleukin-1alpha) treated cells. A) Human articular chondrocytes were treated with 100 U/ml IL-1α (Interleukin-1alpha) and/or 200 U/ml TNFα (Tumor Necrosis Factor alpha) for 24 hours. Western blot analysis of chondrocyte conditioned media with IL-6 (Interleukin-6), IL-8 (Interleukin-8) antibodies; B) Cells were pretreated 24 hours with 25 μM MGDG before treatment with IL-1α for 24 hours. Upper panels: Western blot of chondrocyte lysates with COX-2 (Cyclooxygenase-2) antibody. Actin was blotted as an internal control. Lower panels: Western blot of conditioned media with IL-6 and IL-8 antibodies; C-D) Quantitation of the repression of IL-6 and IL-8 by MGDG. The average of densitometric analysis of Western blots performed in six determinations on three primary cultures of human chondrocytes in duplicate dishes. To show the repression by MGDG the % value referred to the value in IL-1α induced cells (100%) is calculated. E, F) Real Time RT-PCR analysis of Collagen II and Collagen I mRNA expression in human articular chondrocytes treated with 100 U/ml IL-1α in the absence and in the presence of MGDG at different concentrations. Insets show Western blot analysis of chondrocyte conditioned media with Collagen type II and Collagen type I antibodies. 15ΔPGJ2, 15-deoxy-Δ12,14-prostaglandin J2; COX-2, cyclooxygenase-2; DGDG, digalactosyldiacylglycerol; IL-1, interleukin-1; IL-6, interleukin-6; IL-8, interleukin-8; MGDG, monogalactosyldiacylglycerol; mPGES, microsomal PGE synthase; NF-kB, nuclear factor-kappaB; P1, cell passage number1; p38, p38 mitogen activated protein kinase; PGE2, prostaglandin E2; TNFα, tumor necrosis factor alpha.