earticle

영어

Gangliosides, play important regulatory roles in cell adhesion, survival and immunosuppressive activity. In this study, we investigated whether gangliosides can affect to immune responses in lipopolysaccharide (LPS)-stimulated porcine kidney cell line, PK15. Cell proliferation decreased, whefreas nitro oxide (NO) increased in 5 ㎍/ml of LPS-stimulated PK15. In high-performance thin layer chromatography (HPTLC) and immunofluorsence analysis showed that gangliosides GM3, GM2, GD3, GD1a and GD1b were detected, however, expression of GM3 and GM2 decreased in LPS-stimulated PK15. Furthermore, we knock- downed UDP-glucose:ceramide glucosyltransferase (Ugcg) by short hairpin (sh) RNA, which catalyzes the initial step in glycosphingolipids biosynthesis pathway. Western blot and HPTLC analyse showed that Ugcg protein and gangliosides expression decreased in Ugcg shRNA infected PK15. When we stimulated Ugcg gene knock-downed PK15 with LPS (5 ㎍/ml), cell proliferation was decreased, however, NO release increased compared to un-infected PK15. Moreover, we investigated the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and tumor necrosis factor-α (TNF-α), and these protein expression levels increased in LPS-stimulated Ugcg gene knock-downed PK15. These results suggest that gangliosides, specifically GM3 and GM2, may play a role in the interaction with components of the inflammatory response, and to be relevant for designing future therapeutic strategies intended to prolong xenotransplantation.