ABSTRACT
The present study was conducted to investigate if the mechanism of human heme oxygenase-1 (HO-1) mediated angiogenesis was through the induction of vascular endothelial growth factor (VEGF). Also, the effect of HO-1 on the expression of transforming growth factor beta (TGF-beta),was studied in the presence and absence of HO-1 inducers. Rat lung microvessel endothelial cell line transduced with human HO-1 gene was subjected to cell culture (six separate experiments). mRNA extraction and reverse transcriptase polymerase chain reaction (RT-PCR) experiments, were performed to evaluate the expression of HO-1, VEGF, and TGF-beta in the presence and absence of HO inducers including H(2)O(2), endotoxin and snake venom metalloproteinase with disintegrin like activity(SnMP). ELISA technique was performed to evaluate the levels of the studied growth factors. The results of the study showed over expression of VEGF in endothelial cells transduced with HO-1 compared to control non-transduced endothelial cells. On the other hand, the expression of TGF-beta and its protein level were markedly inhibited in HO-1 transduced endothelial cells compared to control non-transduced cells. Endotoxin and SnMP showed more prominent effect on the expression of VEGF and suppression of TGF-beta in HO-1 transduced endothelial cells, suggesting that their effect is most probably mediated through induction of HO-1.
Subject(s)
Endothelium, Vascular/metabolism , Gene Transfer Techniques , Heme Oxygenase (Decyclizing)/genetics , Retroviridae/genetics , Animals , Blotting, Western , Capillaries/physiology , Cells, Cultured , Cytokines/metabolism , Endothelial Growth Factors/metabolism , Endotoxins/metabolism , Enzyme-Linked Immunosorbent Assay , Heme Oxygenase-1 , Humans , Hydrogen Peroxide/pharmacology , Intercellular Signaling Peptides and Proteins/metabolism , Lymphokines/metabolism , Membrane Proteins , Neovascularization, Physiologic , RNA/metabolism , RNA, Messenger/metabolism , Rats , Reverse Transcriptase Polymerase Chain Reaction , Transforming Growth Factor beta/metabolism , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Nineteen patients with juvenile chronic arthritis (JCA), ten with systemic (s)-JCA, and nine with polyarticular-onset (p)-JCA were examined for interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, IL-2R, and IL-10 levels. Power Doppler sonography (PDS) for the more affected knee was used in all of them to evaluate soft tissue vascularity. Serum levels of IL-6 were significantly higher in JCA patients than in controls (P<0.007). Patients with p-JCA showed higher levels of IL-6 than patients with s-JCA, and the difference was statistically nonsignificant. Serum IL-6 levels in all patients correlated significantly with the degree of vascularity detected by PDS (P<0.01). This correlation was more pronounced in p-JCA patients (P<0.01 in p-JCA vs P<0.05 in s-JCA). Serum levels of TNF-alpha were higher in patients with JCA than in controls (P<0.0001). Serum levels of TNF-alpha were significantly greater in patients with s-JCA than in p-JCA (P=0.008). Soluble IL-2R levels were higher in patients with JCA than controls (P<0.0002). Serum levels of IL-2R correlated significantly with pannus thickness in p-JCA (P<0.01) and inversely with methoxetrate (MTX) duration in s-JCA (P<0.05). Serum levels of IL-10 were significantly higher in JCA patients than in controls ( P<0.0008). Serum IL-10 levels in all patients correlated significantly inversely with hemoglobin levels (r=-0.50, P<0.05), total leukocytic count (TLC) (r=-0.58, P<0.01), and intra-articular steroid injection (r=+0.56, P<0.01). In s-JCA, IL-10 levels correlated significantly with MTX weekly dose ( P<0.05). In conclusion, a significant correlation of serum IL-6 levels with the degree of knee joint vascularity was found, and this correlation was more pronounced in p-JCA, which may stress the role of IL-6 as an inducer of neoangiogenesis in JCA.
