title: Recombinant protein glutathione S-transferases P1 attenuates inflammation in mice.
abstract: We have reported that intracellular glutathione S-transferases P1 (GSTP1) suppresses LPS (lipopolysaccharide)-induced excessive production of pro-inflammatory factors by inhibiting LPS-stimulated MAPKs (mitogen-activated protein kinases) as well as NF-kappaB activation. But under pathogenic circumstances, physiologic levels of GSTP1 are insufficient to stem pro-inflammatory signaling. Here we show that LPS-induced up-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW246.7 cells is significantly reduced by incubating cells with recombinant GSTP1 protein. In vivo study demonstrates that treatment of mice (i.p.) with recombinant GSTP1 protein effectively suppresses the devastating effects of acute inflammation, which includes reduction of mortality rate of endotoxic shock, alleviation of LPS-induced acute lung injury and abrogation of thioglycolate (TG)-induced peritoneal deposition of leukocytes and polymorphonuclear cells (PMNs). Meanwhile, GSTP1 prevented LPS-induced TNF-alpha, IL-1beta, MCP-1 and NO production. Further investigation by using confocal microscopy and flow cytometry shows that recombinant GSTP1 protein can be delivered into RAW246.7 cells, mouse peritoneal macrophages and HEK 293 cells suggesting that extracellular GSTP1 protein could be transported across plasma membrane and act as a cytosolic protein. In conclusion our research demonstrates a new finding that increasing cellular GSTP1 level by supplement of recombinant GSTP1 effectively suppresses the devastating effects of acute inflammation.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210093, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Luo
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.forename: Qin
authors.initials: Q
authors.lastname: Feng
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Bin
authors.initials: B
authors.lastname: Xue
authors.forename: Jiayin
authors.initials: J
authors.lastname: Shen
authors.forename: Yin
authors.initials: Y
authors.lastname: Ye
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Han
authors.forename: Hsiaoyen
authors.initials: H
authors.lastname: Ma
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Yin
mesh_terms: D055371:Acute Lung Injury / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology; D000818:Animals; D002460:Cell Line; D018932:Chemokine CCL2 / Q000276:immunology; D051549:Glutathione S-Transferase pi / Q000276:immunology / Q000494:pharmacology*; D006801:Humans; D007249:Inflammation / Q000178:diet therapy / Q000276:immunology; D053583:Interleukin-1beta / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology / Q000633:toxicity; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000276:immunology; D009569:Nitric Oxide / Q000276:immunology; D052247:Nitric Oxide Synthase Type II / Q000276:immunology; D021381:Protein Transport / Q000187:drug effects / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology; D012772:Shock, Septic / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
chemical_list: C111835:CCL2 protein, human; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; C496557:Gstp1 protein, mouse