Proteins　N-glycosylation　is　significantly　increased　in　the　activated　human　hepatic　stellate　cells　(HSCs)　stimulated　by　transforming　growth　factor-beta　1　(TGF-beta　1)　compared　to　the　quiescent　HSCs　according　to　our　previous　study.　However,　little　is　known　about　the　alteration　of　N-glycoprotein　profiles　in　the　activated　HSCs.　Profiles　of　N-glycopeptides　/　N-glycoproteins　/　N-glycosites　in　LX-2　cells,　with　and　without　activation　by　TGF-beta　1,　were　identified　and　compared　using　hydrazide　chemistry　enrichment　coupled　with　liquid　chromatography　-　mass　spectrometry　analysis.　Western　blot　and　immunohistochemistry　were　further　used　for　validation.　A　total　of　103　non-redundant　N-glycopeptides,　with　107　glycosylation　sites　from　86　N-glycoproteins,　were　identified　in　activated　and　quiescent　LX-2　cells　respectively.　Among　these,　23　proteins　were　known　N-glycoproteins,　and　58　were　newly　identified　N-glycoproteins.　In　addition,　43　proteins　(e.g.,　pigment　epithelium-derived　factor　and　clathrin　heavy　chain　1)　were　solely　identified　or　up-regulated　in　the　activated　LX-2　cells,　which　participated　in　focal　adhesion　and　glycosaminoglycan　degradation　pathways　and　were　involved　in　interaction　clusters　of　cytoskeletal　proteins　(e.g.,　myosin　light　chains　and　keratins).　The　increased　expression　of　glucosamine　(N-acetyl)-6-sulfatase　and　phospholipase　C　beta　2　and　the　decreased　expression　of　zinc　finger　and　BTB　domain-containing　protein　1　were　validated　in　the　activated　compared　to　the　quiescent　LX-2　cells.　In　conclusion,　increased　expression　of　N-glycoproteins　and　N-glycosites　play　important　roles　in　cellular　contractility,　signal　transduction,　and　responses　to　stimuli　in　the　activated　HSCs,　which　might　provide　useful　information　for　discovering　novel　molecular　mechanism　of　HSC　activation　and　therapeutic　targets　in　liver　fibrosis.