TNP-470, an angiogenesis inhibitor, suppresses the progression of peritoneal fibrosis in mouse experimental model

Department of Histology and Cell Biology, Nagasaki University, Nagasaki, Nagasaki, Japan
Kidney International (Impact Factor: 8.56). 11/2004; 66(4):1677-85. DOI: 10.1111/j.1523-1755.2004.00935.x
Source: PubMed


In patients on long-term peritoneal dialysis (PD), angiogenesis and vasculopathy are observed in the peritoneum, and the degree of vascularization correlates with the area of fibrotic tissue, suggesting the involvement of angiogenesis in the progression of peritoneal fibrosis. The aim of the present study was to evaluate the effect of TNP-470, an anti-angiogenic compound, on the development of peritoneal fibrosis induced by chlorhexidine gluconate (CG).
Peritoneal fibrosis was induced by injection of CG into peritoneal cavity of Institute for Cancer Research (ICR) mice. TNP-470 was injected subcutaneously with CG. Mice were sacrificed, and peritoneal tissues were dissected out at days eight and 16 after CG and TNP-470 injection. The expression patterns of CD31 (as a marker of endothelial cells), vascular endothelial cell growth factor (VEGF), alpha-smooth muscle actin (as a marker of myofibroblasts), heat shock protein 47 (HSP47), type III collagen, F4/80 (as a marker of mice macrophages), proliferating cell nuclear antigen (PCNA), and cyclin-dependent kinase 2 (Cdk2) were examined by immunohistochemistry.
CG-injected mice showed thickening of the submesothelial zone and increased number of vessels, myofibroblasts, and infiltrating macrophages. The expression levels of VEGF, type III collagen, and HSP47 were increased, and a large number of PCNA-positive cells and Cdk2-expressing cells were observed in the thickened submesothelial area. Treatment with TNP-470 suppressed the submesothelial zone thickening and reduced collagen III expression as well as angiogenesis. TNP-470 also decreased the number of VEGF-expressing cells, myofibroblasts, macrophages, PCNA-positive cells, and Cdk2-expressing cells.
Our results indicate the involvement of angiogenesis in the progression of peritoneal fibrosis, and suggest that TNP-470 may be potentially useful for the prevention of peritoneal fibrosis through inhibition of angiogenesis and suppression of myofibroblast proliferation.

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    • "For example, expression of VEGF in mesothelial cells that have undergone epithelial-to-mesenchymal transition is increased in PD patients (Aroeira et al., 2005); a large number of VEGFpositive cells were observed in the thickening submesothelial area (Yoshio et al., 2004); and treatment with a neutralizing anti- VEGF monoclonal antibody was able to prevent structural and functional microvascular alterations induced by hyperglycemia (De Vriese et al., 2001). As such, inhibition of VEGF expression may be an interesting therapeutic approach for prevention of peritoneal fibrosis. "
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    ABSTRACT: Peritoneal fibrosis is one of serious complications in patients with peritoneal dialysis (PD), and associated with the loss of peritoneal membrane ultrafiltration function. In this study, we investigated whether suramin, an inhibitor that blocks multiple growth factors with their receptors, would prevent development of peritoneal fibrosis in a rat model. Rats were given a daily intraperitoneal injection of chlorhexidine gluconate for three weeks to induce peritoneal fibrosis. Administration of suramin at 5, 10, 20 mg/kg dose-dependently attenuated peritoneal membrane thickening and expression of collagen I, fibronectin and α-smooth muscle actin. Increased expression of TGF-β1 and phosphorylation of Smad3 were detected in fibrotic peritoneum, and inhibited by suramin treatment. Suramin was also effective in blocking chlorhexidine gluconate-induced phosphorylation of IκB and NF-κB p65, expression of several inflammatory cytokines and infiltration of macrophages in the peritoneum. Moreover, suramin suppressed expression of vascular endothelial growth factor, a molecule associated with angiogenesis in the injured peritoneum. Therefore, our results indicate that suramin treatment can effectively alleviate the development of peritoneal fibrosis by suppression of TGF-β1 signaling, inflammation, angiogenesis and suggest that suramin may have therapeutic potential for prevention of peritoneal fibrosis in PD patients.
    Journal of Pharmacology and Experimental Therapeutics 08/2014; 351(2). DOI:10.1124/jpet.114.215228 · 3.97 Impact Factor
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    • "Peritoneal fibrosis was induced by intraperitoneal injection of 0.05% CG in 15% ethanol dissolved in saline, as described previously with a slight modification [3, 36]. Mice received injections of CG into the peritoneal cavity at a volume of 10 ml/kg body weight 3 times a week for 3 weeks. "
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    ABSTRACT: Leptin is a hormone mainly produced by white adipose cells, and regulates body fat and food intake by acting on hypothalamus. Leptin receptor is expressed not only in the hypothalamus but in a variety of peripheral tissues, suggesting that leptin has pleiotropic functions. In this study, we investigated the effect of leptin on the progression of peritoneal fibrosis induced by intraperitoneal injection of chlorhexidine gluconate (CG) every other day for 2 or 3 weeks in mice. This study was conducted in male C57BL/6 mice and leptin-deficient ob/ob mice. Peritoneal fluid, blood, and peritoneal tissues were collected 15 or 22 days after CG injection. CG injection increased the level of leptin in serum and peritoneal fluid with thickening of submesothelial compact zone in wild type mice, but CG-injected ob/ob mice attenuate peritoneal fibrosis, and markedly reduced the number of myofibroblasts, infiltrating macrophages, and blood vessels in the thickened submesothelial area. The 2-week leptin administration induced a more thickened peritoneum in the CG-injected C57BL/6 mice than in the PBS group. Our results indicate that an upregulation of leptin appears to play a role in fibrosis and inflammation during peritoneal injury, and reducing leptin may be a therapeutically potential for peritoneal fibrosis.
    Acta histochemica et cytochemica official journal of the Japan Society of Histochemistry and Cytochemistry 04/2013; 46(2):75-84. DOI:10.1267/ahc.13005 · 1.39 Impact Factor
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    • "Negative controls were performed by omitting the primary antibody. Finally, PCRI was determined using the procedure of Kang et al [17], and the number of PCNA-positive cells were counted in 10 non-overlapping sequential fields at 400×magnification [18]. "
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    ABSTRACT: The objective of this study was to investigate the role of endothelial progenitor cells (EPCs) in the modulation of ischemia-reperfusion injury (IRI) in a partial nephrectomy (PN) rat model using early-phase ischemic preconditioning (IPC). Ninety male Sprague-Dawley rats were randomly divided into three groups following right-side nephrectomy: Sham-operated rats (surgery without vascular clamping); PN rats (renal blood vessels were clamped for 40 min and PN was performed); and IPC rats (pretreated with 15 min ischemia and 10 min reperfusion). At 1, 3, 6, 12, 24 h, and 3 days after reperfusion, the pool of circulating EPCs and kidneys were harvested. The extent of renal injury was assessed, along with EPC number, cell proliferation, angiogenesis, and vascular growth factor expression. Pretreated rats exhibited significant improvements in renal function and morphology. EPC numbers in the kidneys were increased at 12 h following reperfusion in the IPC group as compared to the PN or Sham groups. Cell proliferation (including endothelial and tubular epithelial cells) and angiogenesis in peritubular capillaries were markedly increased in kidneys treated with IPC. In addition, vascular endothelial growth factor-A (VEGF-A) and stromal cell-derived factor-1α (SDF-1α) expression in the kidneys of pretreated rats was increased compared to rats subjected to PN. OUR INVESTIGATION SUGGESTED THAT: (1) the early phase of IPC may attenuate renal IRI induced by PN; (2) EPCs play an important role in renal protection, involving promotion of cell proliferation and angiogenesis through release of several angiogenic factors.
    PLoS ONE 01/2013; 8(1):e55389. DOI:10.1371/journal.pone.0055389 · 3.23 Impact Factor
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