Angiogenic effects of stromal cell-derived factor-1 (SDF-1/CXCL12) variants in vitro and the in vivo expressions of CXCL12 variants and CXCR4 in human critical leg ischemia
Angiogenic effects of stromal cell-derived factor-1 (SDF-1/CXCL12) variants in vitro and the in vivo expressions of CXCL12 variants and CXCR4 in human critical leg ischemia
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DOI：
10.1016/j.jvs.2009.10.044
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发表时间：
2010-03-01
影响因子：
4.3
通讯作者：
Baker, Daryll M.
中科院分区：
文献类型：
--
作者：
Ho, Teik K.;Tsui, Janice;Baker, Daryll M.
目的：下肢严重缺血（CLI）具有较高的发病率和死亡率。治疗性血管生成作为一种可能的替代治疗方案仍在研究中。CXCL12是一种趋化因子，已知有两个剪接变体，CXCL12 α和CXCL12 β，但其意义尚不清楚。本研究探讨了CXCL12在人CIA中的血管生成作用、CXCL12蛋白表达及受体CXCR4。方法：采用体外培养的人微血管内皮细胞（HMEC-1）。亚甲基蓝法和细胞计数法检测细胞增殖情况。4′-6-二氨基-2-苯基吲哚染色后计数缩缩细胞核，caspase-3法证实细胞凋亡。毛细管形成实验采用基质凝胶法。Western blotting检测信号通路活性。在体内，对CLI患者和对照组的下肢进行腓肠肌活检（n = 12）。然后进行免疫组织化学、双免疫荧光标记和免疫印迹。结果：CXCL12抑制HMEC-1细胞凋亡（P < 0.01），促进HMEC-1细胞增殖（P < 0.05）和毛细血管形成（P < 0.01）。与CXCL12 α相比，CXCL12 β对细胞凋亡和细胞增殖的影响更大（P < 0.01）。用这两种变体治疗导致PI3K/Akt和p44/42的时间依赖性激活，而不是p38 MAP激酶。在CLI中，骨骼肌纤维表达CXCL12 α，而CXCL12 β的表达极少。CXCR4广泛表达并在微血管中进行了共定位。在CLI中，CXCL12 α和CXCR4的表达增加了2.6倍（P < 0.01），而CXCL12 β的表达没有增加（P < 0.05）。结论：该研究表明CXCL12 β具有更强的血管生成特性，但在人类CLI活检中没有升高。这为CXCL12变异在CLI病理生理血管生成反应中的作用提供了所有有趣的发现。[J] .中华外科杂志，2010;51:689-99。
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, CXCL12 alpha and CXCL12 beta, but the significance remains unknown. The study investigated the angiogenic effects of CXCL12, protein expressions of CXCL12, and the receptor CXCR4 in human CIA.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 immunoflorescence 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 CXCL12 alpha, CXCL12 beta has a greater effect oil 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 CXCL12 alpha was expressed by skeletal muscle fibers with minimal expression of CXCL12 beta. CXCR4 was extensively expressed and colocalized to microvessels. A significant 2.6-fold increase in CXCL12 alpha and CXCR4 expressions (P < .01) were noted in CLI but not for CXCL12 beta (P > .05).Conclusions:The study showed that CXCL12 beta had more potent angiogenic properties but was not elevated ill human CLI biopsies. This provided all interesting finding oil the role of CXCL12 variants in pathophysiologic angiogenic response in CLI. (J Vasc Surg 2010;51:689-99.)