Ryan Peter L, Youngblood Ramey C, Harvill Jane, Willard Scott T
Department of Animal and Dairy Sciences, College of Agriculture and Life Sciences, Mississippi State University, Mississippi State, Mississippi 39762, USA.
Ann N Y Acad Sci. 2005 May;1041:398-414. doi: 10.1196/annals.1282.061.
Relaxin is known to promote vascular endothelial growth factor (VEGF) expression in reproductive tissue, and successful wound healing depends on good vascularization of wound sites, a process that relaxin may facilitate. Thus, the objective of this study was to evaluate the efficacy of relaxin on the development of vascular tissue at wound sites in a novel VEGF receptor 2-luc (VEGFR2-luc) transgenic mouse wound model by monitoring the rate of VEGFR2-luc-mediated gene expression using bioluminescence and real-time imaging. To this end, 12 FVB/N VEGFR2-luc transgenic male mice were assigned to treatments (six per group): saline alone or relaxin (1 g/6 h/14 days) administered intraperitoneally (i.p.). On day 0, a set of full-thickness wounds (6-mm punch) were generated under anesthesia on the dorsal aspect of each mouse. Photonic emissions were recorded (5-min collection of photons) from wound sites 10 min after the administration of luciferin (150 mg/kg i.p.) on day 0 and on days 1, 2, 4, 7, 9, 11, and 14 postwounding to quantify luciferase activity using an IVIS 100 biophotonic imaging system. Animals were sacrificed (three per group) on day 7 or 14, and wound tissue specimens were recovered for molecular and histologic analyses. Although photonic emission from wound sites increased (P < .001) over time with peak values obtained by day 7, no significant (P > .05) effect of relaxin treatment on VEGFR2-luc gene expression was noted at wound sites. Whereas measuring relaxin's effect on angiogenesis indirectly via the VEGFR2 model was not successful, photonic imaging provides an exciting new tool using alternative models (i.e., VEGF-luc mouse) to study relaxin-induced gene expression in normal (i.e., wound healing) or tumorigenic tissues in real time.
已知松弛素可促进生殖组织中血管内皮生长因子(VEGF)的表达,而伤口的成功愈合取决于伤口部位良好的血管形成,松弛素可能会促进这一过程。因此,本研究的目的是通过使用生物发光和实时成像监测VEGFR2-luc介导的基因表达速率,评估松弛素对新型VEGF受体2-荧光素酶(VEGFR2-luc)转基因小鼠伤口模型中伤口部位血管组织发育的功效。为此,将12只FVB/N VEGFR2-luc转基因雄性小鼠分配至各处理组(每组6只):单独注射生理盐水或腹腔注射(i.p.)松弛素(1 g/6小时/14天)。在第0天,在每只小鼠背部麻醉状态下造成一组全层伤口(6毫米打孔)。在第0天以及伤口形成后的第1、2、4、7、9、11和14天,在腹腔注射荧光素(150 mg/kg i.p.)10分钟后,从伤口部位记录光子发射(5分钟收集光子),以使用IVIS 100生物光子成像系统量化荧光素酶活性。在第7天或第14天处死动物(每组3只),并回收伤口组织标本进行分子和组织学分析。尽管伤口部位的光子发射随时间增加(P < 0.001),在第7天达到峰值,但未观察到松弛素处理对伤口部位VEGFR2-luc基因表达有显著(P > 0.05)影响。虽然通过VEGFR2模型间接测量松弛素对血管生成的作用未成功,但光子成像提供了一种令人兴奋的新工具,可使用替代模型(即VEGF-luc小鼠)实时研究松弛素在正常(即伤口愈合)或致瘤组织中诱导的基因表达。