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目的:探讨动静脉血管吻合口处再狭窄的机制,明确缺氧诱导血管平滑肌的缺氧诱导因子-1α(HIF-1α)表达与细胞增殖的关系。方法:52只新西兰大白兔行髂动静脉吻合术,随机分入术后常氧组(21%氧)、术后给氧组(30%氧)和假手术组。ELISA法检测兔血浆血小板源性生长因子B(PDGF-B)表达,Western blotting检测吻合口组织HIF-1α、PDGF-B、ERK1/2、Akt表达。以不同处理组的兔血浆培养兔主动脉平滑肌细胞,BrdU增殖试验检测细胞增殖,Western blotting检测细胞中HIF-1α、PDGF-B、ERK1/2、Akt表达。结果:术后常氧组兔血浆中PDGF-B和吻合口组织HIF1-α、PDGF-B、ERK1/2的表达明显增高。术后常氧组兔血浆培养的平滑肌细胞增生显著,其增生效应可被PDGF受体诺氨酸激酶阻滞剂AG1259阻断,缺氧条件下细胞内HIF1-α、PDGF-B、ERK1/2表达均明显增高。结论:局部缺氧是导致动静脉吻合口组织平滑肌细胞增殖、血管再狭窄发生的重要因素,术后氧疗是降低再狭窄发生的重要方法。
Objective: To investigate the mechanism of restenosis at arteriovenous anastomosis and to clarify the relationship between hypoxia-inducible factor-1α (HIF-1α) and cell proliferation induced by hypoxia in vascular smooth muscle. Methods: Fifty - two New Zealand white rabbits underwent iliac arteriovenous anastomosis and were randomly divided into normoxia group (21% oxygen), postoperative oxygen group (30% oxygen) and sham operation group. The expression of platelet-derived growth factor B (PDGF-B) in rabbit plasma was detected by ELISA. The expressions of HIF-1α, PDGF-B, ERK1 / 2 and Akt were detected by Western blotting. Rabbit aorta smooth muscle cells were cultured in different treatment groups. BrdU proliferation assay was used to detect cell proliferation. The expression of HIF-1α, PDGF-B, ERK1 / 2 and Akt was detected by Western blotting. Results: The expressions of HIF1-α, PDGF-B and ERK1 / 2 in PDGF-B and anastomotic tissue in the normoxia group were significantly increased after operation. Proliferation of smooth muscle cells in normoxia rabbits was significantly increased after operation, and its proliferation was blocked by AG1259, a PDGF receptor receptor antagonist. HIF-1α, PDGF-B, ERK1 / 2 Expression were significantly higher. Conclusion: Hypoxia is an important factor leading to the proliferation of vascular smooth muscle cells and vascular restenosis in arteriovenous anastomosis. Postoperative oxygen therapy is an important method to reduce the occurrence of restenosis.