The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, ResourceTM S column chromatography and reverse-phase high performance l...The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, ResourceTM 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 c DNA encoding the precursor of the compound was cloned from the c DNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α(TNF-α), interferon ?(IFN-?), 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-?, IFN-?, 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.展开更多
The present study was designed to search for compounds with analgesic activity from the Schizophyllum commune(SC), which is widely consumed as edible and medicinal mushroom world. Thin layer chromatography(TLC), tosil...The present study was designed to search for compounds with analgesic activity from the Schizophyllum commune(SC), which is widely consumed as edible and medicinal mushroom world. Thin layer chromatography(TLC), tosilica gel column chromatography, sephadex LH 20, and reverse-phase high performance liquid chromatography(RP-HPLC) were used to isolate and purify compounds from SC. Structural analysis of the isolated compounds was based on nuclear magnetic resonance(NMR). The effects of these compounds on voltage-gated sodium(NaV) channels were evaluated using patch clamp. The analgesic activity of these compounds was tested in two types of mouse pain models induced by noxious chemicals. Five phenolic acids identified from SC extracts in the present study included vanillic acid, m-hydroxybenzoic acid, o-hydroxybenzeneacetic acid, 3-hydroxy-5-methybenzoic acid, and p-hydroxybenzoic acid. They inhibited the activity of both tetrodotoxin-resistant(TTX-r) and tetrodotoxin-sensitive(TTX-s) NaV channels. All the compounds showed low selectivity on NaV channel subtypes. After intraperitoneal injection, three compounds of these compounds exerted analgesic activity in mice. In conclusion, phenolic acids identified in SC demonstrated analgesic activity, facilitating the mechanistic studies of SC in the treatment of neurasthenia.展开更多
The present study was designed to investigate the antimalarial activity of synthetic hepcidin and its effect on cytokine secretion in mice infected with Plasmodium berghei.The mice were infected with P.berghei intrave...The present study was designed to investigate the antimalarial activity of synthetic hepcidin and its effect on cytokine secretion in mice infected with Plasmodium berghei.The mice were infected with P.berghei intravenously and treated with hepcidin according to 4-day suppression test and Rane's test.The serum levels of interleukins(IL-1β,IL-2,IL-6,IL-10,IL-12p70,and IL-17A),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ) in the experimental mice were determined using a cytometric bead array(CBA)kit.The survival rate of the infected mice was also registered.Additionally,the serum iron,alanine transaminase(ALT),aspartate transaminase(AST),and total bilirubin(BIL) were detected to evaluate liver functions.Hepcidin exerted direct anti-malarial function in vivo and increased survival rate in a dose-dependent manner.In addition,the secretion of T helper cell type 1(Th1),Th2,and Th17 cytokines,TNF-α,and IFN-γ were inhibited by hepcidin.In conclusion,our results demonstrated that synthetic hepcidin exerts in vivo antimalarial activity and possesses anti-inflammatory function,which provides a basis for future design of new derivatives with ideal anti-malarial activity.展开更多
基金supported by Ministry of Science and Technology of China(Nos.2013CB911300,2010CB529800)the National Science Foundation of China(Nos.U1132601,31200590)the National Science Foundation of Yunnan Province(No.P0120150010)
文摘The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, ResourceTM 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 c DNA encoding the precursor of the compound was cloned from the c DNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α(TNF-α), interferon ?(IFN-?), 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-?, IFN-?, 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.
基金partly supported by Ministry of Science and Technology of China(No.2013CB911300)National Natural Science Foundation of China(Nos.81225024,31025025,31260208,and U1132601)
文摘The present study was designed to search for compounds with analgesic activity from the Schizophyllum commune(SC), which is widely consumed as edible and medicinal mushroom world. Thin layer chromatography(TLC), tosilica gel column chromatography, sephadex LH 20, and reverse-phase high performance liquid chromatography(RP-HPLC) were used to isolate and purify compounds from SC. Structural analysis of the isolated compounds was based on nuclear magnetic resonance(NMR). The effects of these compounds on voltage-gated sodium(NaV) channels were evaluated using patch clamp. The analgesic activity of these compounds was tested in two types of mouse pain models induced by noxious chemicals. Five phenolic acids identified from SC extracts in the present study included vanillic acid, m-hydroxybenzoic acid, o-hydroxybenzeneacetic acid, 3-hydroxy-5-methybenzoic acid, and p-hydroxybenzoic acid. They inhibited the activity of both tetrodotoxin-resistant(TTX-r) and tetrodotoxin-sensitive(TTX-s) NaV channels. All the compounds showed low selectivity on NaV channel subtypes. After intraperitoneal injection, three compounds of these compounds exerted analgesic activity in mice. In conclusion, phenolic acids identified in SC demonstrated analgesic activity, facilitating the mechanistic studies of SC in the treatment of neurasthenia.
基金supported by the grants from China National Natural Science Foundation of Young Scientists(No.31201717)Special Foundation for Young Scientists of Jiangsu Province(No.BK2012365)Jiangsu Province Science and Technology Support Project(No.BE2012748)
文摘The present study was designed to investigate the antimalarial activity of synthetic hepcidin and its effect on cytokine secretion in mice infected with Plasmodium berghei.The mice were infected with P.berghei intravenously and treated with hepcidin according to 4-day suppression test and Rane's test.The serum levels of interleukins(IL-1β,IL-2,IL-6,IL-10,IL-12p70,and IL-17A),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ) in the experimental mice were determined using a cytometric bead array(CBA)kit.The survival rate of the infected mice was also registered.Additionally,the serum iron,alanine transaminase(ALT),aspartate transaminase(AST),and total bilirubin(BIL) were detected to evaluate liver functions.Hepcidin exerted direct anti-malarial function in vivo and increased survival rate in a dose-dependent manner.In addition,the secretion of T helper cell type 1(Th1),Th2,and Th17 cytokines,TNF-α,and IFN-γ were inhibited by hepcidin.In conclusion,our results demonstrated that synthetic hepcidin exerts in vivo antimalarial activity and possesses anti-inflammatory function,which provides a basis for future design of new derivatives with ideal anti-malarial activity.
