xpression in macrophages in vitro [29]. Regularly, we also demonstrated that H-LF41 pretreatment for 10 days was related to aggrandizement of LPS-induced hepatic IL-10 protein, although LF41 itself had no stimulatory effect on hepatic IL-10 levels within the absence of LPS (Fig 6A). This amplification impact on hepatic IL-10 levels was abolished after COX-2 blockade (Fig 6A). Furthermore, LF41-mediated alleviation of serum ALT activity ” induced by LPS was abrogated by administration of IL-10-specific antibody 30 minutes before LPS treatment (Fig 6B). Since the isotype control for ” the antibody had no Fig 6. LF41 pretreatment considerably elevated LPS-activated hepatic IL-10, which accounts for the attenuation of serum ALT levels. (A) Mice (LPS-treated groups: n = 80 per group; the remainder: n = six per group) were pretreated PBS or H-LF41 for ten days, either alone or “
22931421“in mixture with celecoxib or its vehicle, and after that provided injection of either PBS or LPS. Four hours following LPS challenge, mice had been killed to test hepatic IL-10 protein levels by ELISA. P 0.05; & P 0.05 compared to H-LF41+LPS; n.s., non-statistical difference. (B) Mice (H-LF41-treated groups: n = ten per group; PBS-treated groups: n = 8 per group) were pretreated with PBS or H-LF41 for 10 days, either singly or in mixture with IL-10-specific antibody (Anti-IL-10) or its isotype handle (Is-IL-10) 30 min prior to LPS inoculation. Serum ALT levels had been determined 16 h just after LPS therapy. P 0.05; + P 0.05 compared to PBS+LPS; & P 0.05 compared to H-LF41+LPS. Values are shown as mean SEM. Results are representative of two experiments with similar results similar effect (Fig 6B), the inhibition of serum ALT expression was indeed dependent on the IL-10. In addition, pretreatment of PGE2 also drastically facilitated hepatic Il10 mRNA levels and reduced serum ALT levels in response to LPS challenge (S3 Fig).Having observed the pivotal role of COX-2 in modulating the upregulation of hepatic PGE2 levels associated with LF41, we investigated the potential other function of COX-2 in LF41-treated mice. Although there was no significant change in intestinal epithelial permeability immediately after oral treatment of H-LF41 for 10 days (Fig 7A), pronounced increase in intestinal permeability was found in LF41-treated mice following blockade of COX-2, but not PGE2 receptor EP4 (Fig 7A). There was no apparent alteration soon after H-LF41 gavage for 10 days inside the expression of pro-inflammatory cytokines (Fig 3A), such as TNF-, IFN-, and IL-1 that can impair intestinal permeability in vitro and in vivo [31]. Therefore, we examined whether the COX-2 blockade would influence the levels of these pro-inflammatory cytokines in order 857290-04-1 LF41-fed mice. Indeed, these mice exhibited enhanced TNF- expression right after the blockade, with improved TNF- 
secretion inside the terminal ileum and upregulated Tn fmRNA levels in both the epithelial Fig 7. Impact of COX-2 or IL-10 blockade on TNF- expression and intestinal permeability in LF41-fed mice. (A)(B) Mice (PBS-treated groups: n = 5 per group; H-LF41-treated groups: n = 7 per group) have been given ten days remedy of PBS or H-LF41, either alone or in mixture with blockade of EP4, COX-2, or IL-10, or co-blockade of TNF- with COX-2 or IL-10, then offered IG inoculation with FITC-Dextran. Three hours later, the FITC-Dextran amount in the blood was determined. I-EP4, EP4-specific inhibitor; automobile, the automobile for celecoxib; celecoxib, COX-2-specific inhibitor; Anti-TNF, TN