Growth hormone down-regulation of Interleukin-1beta and Interleukin-6 induced acute phase protein gene expression is associated with increased gene expression of suppressor of cytokine signal-3.
Severe burn induces the hepatic acute phase response. We previously showed that recombinant human growth hormone (GH) treatment after burn down-regulated acute phase protein (APP) production and gene expression in vivo. In this study, we hypothesized that the inhibitory effect of GH on the hepatic acute phase response was due to increased suppressor of cytokine signaling (SOCS) gene expression. HepG2 cells were treated with Interleukin-1beta (IL-1beta; 2 ng/mL) and interleukin 6 (IL-6; 20 ng/mL) alone or combined with GH (2 microg/mL) for 15 and 30 min, and 1, 2, and 4 h. The levels of gene expression for alpha1-acid glycoprotein (AGP), alpha1-antitrypsin (ATT), and SOCS (CIS, SOCS-1, 2, and 3) were measured by reverse transcript-polymerase chain reaction (RT-PCR). APP levels in the supernatant were determined by enzyme-linked immunosorbent sandwich assay (ELISA). The gene expression of AGP and ATT were also measured in HepG2 cells transfected with pEF-Flag-l/mSOCS-3 plasmid after IL-1beta or IL-6 treatment. Data are expressed as means +/- SEM, and statistical analysis was performed by one- or two-way analysis of variance. IL-1beta and IL-6 induced AGP and ATT gene expression and protein production, respectively, which was down-regulated by GH treatment. SOCS-3 but not CIS, SOCS-1, or SOCS-2 gene expression was significantly increased by GH treatment. APP gene expression was significantly decreased in cells transfected with plasmid over expressing SOCS-3 after IL-6 and IL-1beta treatment. GH attenuates IL-1beta or IL-6 induced APP gene expression, which is associated with increased expression of SOCS-3. This study suggests that SOCS-3 plays an important role in the suppression of cytokine signaling by GH in down-regulating the acute phase response after injury.
Wu
Xiaowu
X
Shriner's Hospitals for Children and Department of Surgery, University of Texas Medical Branch, Galveston, Texas 77550, USA.
Herndon
David N
DN
Wolf
Steven E
SE
D000209:Acute-Phase Proteins / Q000096:biosynthesis* / Q000235:genetics; D000210:Acute-Phase Reaction / Q000188:drug therapy*; D000818:Animals; D002056:Burns / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D017077:Culture Media, Conditioned / Q000737:chemistry; D004268:DNA-Binding Proteins; D005786:Gene Expression Regulation / Q000187:drug effects*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D019382:Human Growth Hormone / Q000494:pharmacology* / Q000627:therapeutic use; D017874:Immediate-Early Proteins / Q000096:biosynthesis / Q000235:genetics; D007375:Interleukin-1 / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008113:Liver Neoplasms / Q000473:pathology; D051379:Mice; D009961:Orosomucoid / Q000096:biosynthesis / Q000235:genetics; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology / Q000627:therapeutic use; D012097:Repressor Proteins; D015398:Signal Transduction / Q000187:drug effects; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014947:Wounds and Injuries / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis / Q000235:genetics; D000093242:Interleukin-6 Inhibitors
D000209:Acute-Phase Proteins; D002352:Carrier Proteins; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D019382:Human Growth Hormone; D017874:Immediate-Early Proteins; D007375:Interleukin-1; D015850:Interleukin-6; D009961:Orosomucoid; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D000515:alpha 1-Antitrypsin; D000093242:Interleukin-6 Inhibitors; C106644:SOCS3 protein, human; C491590:Socs1 protein, mouse; C491409:Socs2 protein, mouse; C491411:Socs3 protein, mouse