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血红素氧合酶-1在胆道缺血再灌注损伤中促胆管周围血管丛再生的作用

Roles of heme oxygenase-1 promoting regeneration of peribiliary vascular plexus in bile duct ischemia/reperfusion injury

摘要:

目的 探讨血红素氧合酶-1(HO-1)在大鼠胆道缺血再灌注损伤中对胆道周围血管丛(PVP)的影响.方法 将128只雄性SD大鼠随机分为盐水组(Saline)、空病毒组(Adv)、诱导组(Adv-HO-1)和抑制组(HO-1 siRNA),每组各32只.分别于术前24 h经阴茎背静脉注射生理盐水、空白腺病毒、H0-1腺病毒和siRNA腺病毒0.5 ml,腺病毒注射剂量为2×109 TU/只.再灌注1h及1、7、14 d检测肝功能,Western blot分析HO-1、血管内皮生长因子(VEGF)、基质细胞衍生因子-1α(SDF-1α)和缺氧诱导因子-1α(HIF-1α)蛋白含量,流式细胞仪检测肝脏和外周血内皮祖细胞(EPCs)比例,进行α-平滑肌肌动蛋白(α-SMA)和血管性血友病因子(vWF)免疫荧光双标染色观察胆管周围的微小血管.结果 手术后诱导组肝功能损伤减轻,诱导HIF-1α、SDF-1α和VEGF的高表达(q =5.68 ~7.52,P<0.01),诱导组术后7d外周血及肝脏中EPCs的比例分别为盐水组的6、9倍(q=12.14、15.26),而抑制组EPCs的比例明显减少(q=4.83、5.07),差异有统计学意义(P<0.01).诱导组术后14 d胆管周围微小血管密度明显高于抑制组.结论 HO-1能诱导HIF-1α、SDF-1α和VEGF的表达,动员EPCs从骨髓向外周血释放,并向肝脏内迁移促进肝内、外PVP损伤的修复和再生.

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abstracts:

Objective To study the effect of heme oxygenase-1 (HO-1) on peribiliary vascular plexus (PVP) in rat bile duct ischemia/reperfusion injury.Methods Total 128 male SD rats were randomly divided into saline group (Saline),empty virus group (Adv),induced group (Adv-HO-1) and suppressed group (HO-1 siRNA),and there were 32 rats in each group.Rats were injected using 0.5 ml of saline,empty adenovirus,HO-1 adenovirus and siRNA adenovirus (2 × 109 TU/rat) via the dorsal penile vein 24 hours before surgery.Liver function was analyzed at 1 hour and 1,7,14 days after reperfusion.HO-1,hypoxiainducible factor-lαt (HIF-1α),stromal cell derived factor-1α (SDF-1α) and vascular endothelial growth factor (VEGF) protein content was analyzed by Western blot.The endothelial progenitor cells (EPCs) ratio in the liver and peripheral blood was detected by flow cytometry.Small vascular around the bile duct was observed by α-smooth muscle actin and yon Willebrand factor double immunofluorescence staining.Results Reduced liver injury and higher expression of HIF-1α,SDF-1α and VEGF in the induced group after surgery (q =5.68-7.52,P <0.01).EPCs ratio in the liver and peripheral blood was significantly higher in the induced group than saline group (q =12.14 and 15.26,P < 0.01),and the suppressed group at 7 days after surgery were less than saline group significantly (q =4.83 and 5.07,P < 0.01).In comparison to the suppressed group,higher density of small vascular around the bile duct was seen in the liver tissue of induced group.Conclusions HO-1 can induce the expression of HIF-1α,SDF-1α and VEGF,and mobilize the release of EPCs to the peripheral from the bone marrow.EPCs migrate to the liver and promote damaged PVP repair and regeneration.

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作者: 黄汉飞 [1] 侯美玲 [1] 许坚吉 [1] 庞天龙 [1] 段键 [1] 李珍 [1] 曾仲 [1] 王昆华 [1]
第一作者: 黄汉飞
期刊: 《中华外科杂志》2014年52卷3期 193-197页 MEDLINEISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.0529-5815.2014.03.009
发布时间: 2014-04-01
基金项目:
云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目
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