PURPOSE: Critical leg ischemia (CLI) is associated with a high morbidity and mortality. Therapeutic angiogenesis is still being investigated as a possible alternative treatment option for CLI. CXCL12, a chemokine, is known to have two spliced variants, CXCL12alpha and CXCL12beta, but the significance remains unknown. The study investigated the angiogenic effects of CXCL12, protein expressions of CXCL12, and the receptor CXCR4 in human CLI. METHODS: In vitro, human microvascular endothelial cells (HMEC-1) were used. Cell proliferation was assessed using methylene blue assay and cell count method. Apoptosis was determined by counting the pyknotic nuclei after 4'-6-diamidino-2-phenylindole staining and confirmed by caspase-3 assay. We employed matrigel as capillary tube formation assay. The activity of signaling pathways was measured using Western blotting. In vivo, gastrocnemius biopsies were obtained from the lower limbs of patients with CLI and controls (n = 12 each). Immunohistochemistry, double immunofluorescence labeling, and Western blotting were then performed. RESULTS: CXCL12 attenuated HMEC-1 apoptosis (P < .01), stimulated cell proliferation (P < .05) and capillary tube formation (P < .01). Compared with CXCL12alpha, CXCL12beta has a greater effect on apoptosis and cell proliferation (P < .01). Treatment with both variants resulted in time-dependent activation of PI3K/Akt and p44/42 but not p38 MAP kinase. In CLI, CXCL12alpha was expressed by skeletal muscle fibers with minimal expression of CXCL12beta. CXCR4 was extensively expressed and colocalized to microvessels. A significant 2.6-fold increase in CXCL12alpha and CXCR4 expressions (P < .01) were noted in CLI but not for CXCL12beta (P > .05). CONCLUSIONS: The study showed that CXCL12beta had more potent angiogenic properties but was not elevated in human CLI biopsies. This provided an interesting finding on the role of CXCL12 variants in pathophysiologic angiogenic response in CLI.
PURPOSE:Critical leg ischemia (CLI) is associated with a high morbidity and mortality. Therapeutic angiogenesis is still being investigated as a possible alternative treatment option for CLI. CXCL12, a chemokine, is known to have two spliced variants, CXCL12alpha and CXCL12beta, but the significance remains unknown. The study investigated the angiogenic effects of CXCL12, protein expressions of CXCL12, and the receptor CXCR4 in human CLI. METHODS: In vitro, human microvascular endothelial cells (HMEC-1) were used. Cell proliferation was assessed using methylene blue assay and cell count method. Apoptosis was determined by counting the pyknotic nuclei after 4'-6-diamidino-2-phenylindole staining and confirmed by caspase-3 assay. We employed matrigel as capillary tube formation assay. The activity of signaling pathways was measured using Western blotting. In vivo, gastrocnemius biopsies were obtained from the lower limbs of patients with CLI and controls (n = 12 each). Immunohistochemistry, double immunofluorescence labeling, and Western blotting were then performed. RESULTS:CXCL12 attenuated HMEC-1 apoptosis (P < .01), stimulated cell proliferation (P < .05) and capillary tube formation (P < .01). Compared with CXCL12alpha, CXCL12beta has a greater effect on apoptosis and cell proliferation (P < .01). Treatment with both variants resulted in time-dependent activation of PI3K/Akt and p44/42 but not p38 MAP kinase. In CLI, CXCL12alpha was expressed by skeletal muscle fibers with minimal expression of CXCL12beta. CXCR4 was extensively expressed and colocalized to microvessels. A significant 2.6-fold increase in CXCL12alpha and CXCR4 expressions (P < .01) were noted in CLI but not for CXCL12beta (P > .05). CONCLUSIONS: The study showed that CXCL12beta had more potent angiogenic properties but was not elevated in human CLI biopsies. This provided an interesting finding on the role of CXCL12 variants in pathophysiologic angiogenic response in CLI.
Authors: Bryan B Edwards; Alexander S Fairman; Jeffrey E Cohen; John W MacArthur; Andrew B Goldstone; Jeffrey B Woo; William Hiesinger; Y Joseph Woo Journal: J Vasc Surg Date: 2015-09-12 Impact factor: 4.268
Authors: Miljan Kuljanin; Gillian I Bell; Stephen E Sherman; Gilles A Lajoie; David A Hess Journal: Diabetologia Date: 2017-07-14 Impact factor: 10.122
Authors: Jeffrey C Gahan; Miguel Gosalbez; Travis Yates; Ezekiel E Young; Diogo O Escudero; Andrew Chi; Michael Garcia-Roig; Ramgopal Satyanarayana; Mark S Soloway; Vincent G Bird; Vinata B Lokeshwar Journal: J Urol Date: 2012-01-15 Impact factor: 7.450
Authors: Miguel Gosalbez; Marie C Hupe; Soum D Lokeshwar; Travis J Yates; John Shields; Muthu K Veerapen; Axel S Merseburger; Charles J Rosser; Mark S Soloway; Vinata B Lokeshwar Journal: J Urol Date: 2013-11-26 Impact factor: 7.450
Authors: Miriam Bobadilla; Neira Sainz; Gloria Abizanda; Josune Orbe; José Antonio Rodriguez; José Antonio Páramo; Felipe Prósper; Ana Pérez-Ruiz Journal: Stem Cells Dev Date: 2014-03-14 Impact factor: 3.272
Authors: Viktoriya Y Rybalko; Chantal B Pham; Pei-Ling Hsieh; David W Hammers; Melissa Merscham-Banda; Laura J Suggs; Roger P Farrar Journal: Biomater Sci Date: 2015-11 Impact factor: 6.843
Authors: Antonius J N M Bastiaansen; Mark M Ewing; Hetty C de Boer; Tineke C van der Pouw Kraan; Margreet R de Vries; Erna A B Peters; Sabine M J Welten; Ramon Arens; Scott M Moore; James E Faber; J Wouter Jukema; Jaap F Hamming; A Yaël Nossent; Paul H A Quax Journal: Arterioscler Thromb Vasc Biol Date: 2013-06-20 Impact factor: 8.311