Platelet-derived growth factor production by cells from Dacron grafts implanted in a canine model.
Platelet-derived growth factor production by cells from Dacron grafts implanted in a canine model.
复制标题
植入犬模型的涤纶移植物细胞产生血小板衍生生长因子。
DOI：
10.1016/s0741-5214(97)70149-6
复制
发表时间：
1997
影响因子：
4.3
通讯作者：
Graham,LM
中科院分区：
文献类型：
--
作者：
Pitsch,RJ;Minion,DJ;Goman,ML;vanAalst,JA;Fox,PL;Graham,LM
目的：先前对犬移植物的研究表明，血小板源性生长因子（PDGF）的产生与内囊厚度相关，呈时间依赖性增加。本研究的目的是确定细胞在血管移植物产生PDGF.MethodsDacron胸腹移植物与自体内皮细胞（ECs），种植在11只比格犬，并删除后4或20周。取移植血管及邻近主动脉培养内皮细胞和平滑肌细胞，放射受体法检测条件培养液中PDGF含量。使用逆转录酶，然后通过聚合酶链反应评估新鲜收获的细胞中的PDGF A链mRNA水平，并表示为甘油醛-3-磷酸脱氢酶信号的比率。结果原代培养的移植物和主动脉内皮细胞在72小时内产生的PDGF量无明显差异，平均分别为368 ± 160和340 ± 81 pg/μ gDNA。原代培养的移植SMC比主动脉SMC产生显著更多的PDGF（分别为584 ± 343和113 ± 94 pg/μg DNA; p < 0.01）。移植物SMC PDGF分泌通过至少六个细胞传代仍然大于主动脉SMC PDGF分泌。主动脉或移植物EC的PDGF A链mRNA水平无显著差异。移植SMC的PDGF A链mRNA水平显著高于主动脉SMC（分别为2.44 ± 0.67和1.45 ± 0.57 pg/μg; p < 0.03）。免疫细胞化学分析在天然主动脉和移植物的EC以及移植物的内皮下SMC中检测到了PDGFA链和B链蛋白，但在天然主动脉的SMC中没有检测到。结论这些结果表明移植物SMC的功能发生了改变，产生的PDGF比主动脉SMC更多。移植物平滑肌细胞产生的PDGF可能参与了内膜增生的发生。（J Vasc Surg 1997;26：70-8.）
PurposePrevious studies of grafts implanted in dogs documented a time-dependent increase in platelet-derived growth factor (PDGF) production that correlated with inner-capsule thickness. The purpose of this study was to identify the cells in vascular grafts that produce PDGF.MethodsDacron thoracoabdominal grafts were seeded with autologous endothelial cells (ECs), implanted in 11 beagles, and removed after 4 or 20 weeks. ECs and smooth muscle cells (SMCs) were cultured from grafts and adjacent aorta, and PDGF in the conditioned media was measured by radioreceptor assay. The PDGF A-chain mRNA level in freshly harvested cells was assessed using reverse transcriptase, followed by polymerase chain reaction, and expressed as a ratio of glyceraldehyde-3-phosphate dehydrogenase signal. Localization of PDGF A-chain and B-chain protein was also examined with immunohistochemical analysis.ResultsGraft and aortic ECs in primary culture did not produce significantly different amounts of PDGF in 72 hours, averaging 368 ± 160 and 340 ± 81 pg/μg DNA, respectively. Graft SMCs in primary culture produced significantly more PDGF than aortic SMCs (584 ± 343 and 113 ± 94 pg/μg DNA, respectively; p < 0.01). Graft SMC PDGF secretion remained greater than aortic SMC PDGF secretion through at least six cell passages. PDGF A-chain mRNA levels were not significantly different for aortic or graft ECs. The PDGF A-chain mRNA level was significantly higher for graft SMCs than aortic SMCs (2.44 ± 0.67 and 1.45 ± 0.57 pg/μg, respectively; p < 0.03). Immunocytochemical analysis detected PDGF A-chain and B-chain protein in the ECs from both native aorta and graft as well as the subendothelial SMCs in the graft, but not in the SMCs of the native aorta.ConclusionsThese results suggest that graft SMCs are functionally altered, producing more PDGF than aortic SMCs. PDGF produced by graft SMCs may contribute to the development of intimal hyperplasia. (J Vasc Surg 1997;26:70-8.)