Identification and characterization of a novel neuropeptide (neuropeptide Y-HS) from leech salivary gland of Haemadipsa sylvestris.
The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, Resource(TM) S column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify the salivary gland extracts (SGE). Structural analysis of isolated compounds was based on Edman degradation and matrix assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cDNA encoding the precursor of the compound was cloned from the cDNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α (TNF-α), interferon γ (IFN-γ), interleukin-6 (IL-6), and monocyte chemotactic protein-1 (MCP-1) were assayed using an enzyme-linked immunosorbent assay (ELISA). The effects on cell proliferation and cell viability were observed using MTT assay. A novel neuropeptide Y (Neuropeptide Y-HS) from the leech salivary gland of H. sylvestris was purified and characterized. It was composed of 36 amino acid residues and the amino acid sequence was determined to be FLEPPERPAVFTSVEQMKSYIKALNDYYLLLGRPRF-NH2, containing an amidated C-terminus. It showed significant inhibitory effects on the production of inflammatory cytokines including TNF-α, IFN-γ, IL-6, and MCP-1. Neuropeptide Y was identified from leeches for the first time. The presence of neuropeptide Y-HS in leech salivary gland may help get blood meal from hosts and inhibit inflammation.
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China.
Wei-Hui
WH
Liu
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China.
Yan
Y
Chen
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China.
Xue-Wei
XW
Bai
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China.
Hui-Min
HM
Yao
Major Cardiovascular Surgery, Kunming Yanan Hospital, Kunming 650051, China.
Xu-Guang
XG
Zhang
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: yanxw@njau.edu.cn.
Xiu-Wen
XW
Yan
Life Sciences College, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: rlai72@njau.edu.cn.
Ren
R
Lai
D000595:Amino Acid Sequence; D000818:Animals; D007155:Immunologic Factors / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D007865:Leeches / Q000737:chemistry*; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000008:administration & dosage* / Q000737:chemistry* / Q000235:genetics; D010449:Peptide Mapping; D012469:Salivary Glands / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D007155:Immunologic Factors; D015850:Interleukin-6; D009478:Neuropeptide Y; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma
Anti-inflammation; Haemadipsa sylvestris; Leech; Neuropeptide Y; Salivary gland