Figure 3
Gal-3 played an important role in the Inflammatory activity of P.g.-LPS. (A) Gal-3 levels in culture media were analysed by ELISA. P.g.-LPS (1 µg/ml) significantly induced Gal-3 in trophoblasts (HTR-8 cells) at 3 days. HTR-8 cells incubated with rhTNF-α (10ng/ml) also significantly produced Gal-3 at 3 days after incubation. (B) TNF-α levels produced from HTR-8 cells with P.g.-LPS (1 μg/ml) were measured. After 3 days, TNF-α production was significantly upregulated. (C,D) HTR-8 cells were pre-treated with or without CAPE (1 μg/ml) for 4 h, pre-exposed to dicumarol/SB203580/U0126 (10 μM) for 30 min, and then grown with or without P.g.-LPS (1 µg/ml) or rhTNF-α (10ng/ml) for 3 days. The Gal-3 or TNF-α concentration in the culture media was analysed by ELISA. CAPE, and U0126 inhibited Gal-3 production induced both by P.g.-LPS and rhTNF-α (C). All inhibitors used were significantly reduced TNF-α secretion from HTR-8 cells at 3 days after LPS-stimulation. Data are presented as the mean ± SD. a and b: significant difference (P < 0.01, p < 0.05) between untreated control, c: significant difference (P < 0.01) between P.g.-LPS/rhTNF-α stimulated and control group; 1-way ANOVA. (E,F) HTR-8 cells were treated with P.g.-LPS (1 μg/ml) with or without a Gal-3 blocking antibody (clone M3/38) or rat control IgG for 3 Days. Gal-3 antibody significantly reduced P.g.-LPS induced TNF-α production (E). COX-2, IL-8, and TNF-α mRNA-expression levels were examined. GAPDH expression was detected as an internal control (F). Data are presented as the mean ± SD. **P < 0.01; 1-way ANOVA. The experiments were performed at least 3 times with similar results.
