title: Induction of chemokines and cytokines before neutrophils and macrophage recruitment in different regions of rat liver after TAA administration.
abstract: Single-dose thioacetamide (TAA) administration induces inflammation and acute liver damage. The mechanism of inflammatory cell recruitment in the liver is still unclear. The aim of this study was to examine the sequence and recruitment of inflammatory cells in different liver regions in relation to CXC- and CC-chemokine and cytokine expression during acute liver injury. Single-dose TAA was administered to rats intraperitoneally, and animals were killed at different time points thereafter. Serum and liver tissue were taken and frozen immediately. Tissue was used for immunostaining cryostat sections, RNA, and protein extraction. RT-PCR and western blotting were performed for RNA and protein analysis, respectively. An early increase (3 h) in CXCL8/IL-8 levels was measured followed by a marked release in MCP1/CCL2 (24 h) serum levels after TAA administration compared with controls. Similarly, an early increase in specific RNA of hepatic chemokines CXCL1/KC and CXCL8/IL-8 was found at 3 h, followed by an upregulation of CXCL5/LIX (6 h), CXCL2/MIP-2 (12 h), and MCP1/CCL2 gene expression at 24-48 h. Further, an induction of pro-inflammatory cytokines IFN-γ and IL-1β followed by IL-6 and TNF-α was observed with a maximum at 12 h. The magnitude of increase in gene expression of TNF-α and MCP1/CCL2 was the highest among all cytokines and chemokines, respectively. By means of immunohistochemistry, an early (12-24 h) increase in the number of only neutrophil granulocytes (NGs) attached to and around portal vessel walls was observed, followed by increased numbers of mononuclear phagocytes (24-48 h) along the sinusoids. Treatment of the human monocytic cell line U-937 with TNF-α increased the gene expression of CXCL1/KC, CXCL8/IL-8, and MCP1/CCL2. Conversely, adding of infliximab (IFX) to the culture medium inhibited this upregulation significantly. In conclusion, single-dose TAA administration induces a sequence of events with a defined upregulation of gene expression of inflammatory chemokines and cytokines and a transient accumulation of NGs within the portal area and macrophages along the sinusoids throughout the liver. Periportal inflammation seems to precede hepatocellular damage.
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Amanzada
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Federico
authors.initials: F
authors.lastname: Moriconi
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Tümen
authors.initials: T
authors.lastname: Mansuroglu
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Cameron
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Giuliano
authors.initials: G
authors.lastname: Ramadori
authors.affiliation: Clinic for Gastroenterology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: Ihtzaz A
authors.initials: IA
authors.lastname: Malik
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism*; D018932:Chemokine CCL2 / Q000097:blood; D018925:Chemokines / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D007150:Immunohistochemistry; D000069285:Infliximab; D010586:Phagocytes / Q000378:metabolism; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013853:Thioacetamide / Q000008:administration & dosage / Q000633:toxicity*; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D018925:Chemokines; D016207:Cytokines; D017931:DNA Primers; D013853:Thioacetamide; D000069285:Infliximab