Macrophages modulate neovascular response in an accute model of inflammation. Expression of pro-angiogenic proteins

DE LA TORRE EULALIA; HOVSEPIAN EUGENIA; GOREN N,; SALES ME

Buenos Aires

Congreso; First French-Argentine Immunology Congress; 2010

Sociedad Argentina de Inmunologia- Sociedad Francesa de Inmunologia

Septic shock (SS) induced by bacterial infections, affect mainly the cardiovascular system. This damage is caused by microorganisms themselves or their endotoxins, and may be enhanced by the infiltration of immune cells in the myocardium. The role of macrophages (MP) in inflammatory angiogenesis during SS is almost unknown. We investigated the angiogenic abitility of peritoneal MP in a model of acute inflammation induced by lippopolysaccharide (LPS) from E. coli in BALB/c mice. We demonstrated that MP are able to stimulate neovascularization at 5x1055 per site, and treatment with LPS (10 ug/ml) in vitro (LPS-MP) reduces MP number required to produce angiogenesis (4x105) potentating this effect (number of skin vessels/mm2) (normal skin: 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. this effect (number of skin vessels/mm2) (normal skin: 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. 5) potentating this effect (number of skin vessels/mm2) (normal skin: 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS. 2) (normal skin: 1.13 E} 0.20, MP: 1.35 E} 0.13, LPS-MP: 2.18 E} 0.19, p <0.01). Furthermore, by Western blot (Wb) (relative O.D.) we observed that LPSMP also increase the expression of pro-angiogenic proteins at the site of inoculation: MMP-9 (MP: 0.36, LPS-MP: 0.64, p <0.05), CD-31 (MP: 0.73, LPS-MP: 1.01, p <0.05) and VEGF-A (MP: 0.13, LPS-MP: 0.73, p <0.05). In vitro, LPS-MP have increased activity of MMP-9 in comparison with MP without treatment (p<0.05) via nuclear factor kB (NF- kB). In addition we studied the role of MP obtained from BALB/c mice treated in vivo with LPS (1 ƒÊg/g, i.p.) (LPSip-MP). We observed an up-regulation of MMP-9 (MP: not detected, LPSip-MP: 1.01, p<0.05) and VEGF-A expression (MP: not detected; LPSip-MP: 1.29, p <0.05) compared to MP from untreated animals. Normal hearts were cultured with LPSip-MP or its supernatants and homogenates showed an increment in proangiogenic molecules expression: CD31 and VEGF-A (p <0.05). We conclude that peritoneal MP are inducers of angiogenesis and infection with LPS enhances their pro-angiogenic effect. In addition, LPS-MP modulates the level of expression of angiogenic molecules in normal hearts and this action enables them as modulators of cardiac angiogenesis during the SS.