VEGF及受体FLK1在大鼠缺血再灌注后肝再生中的表达及变化

发布时间：2018-06-05 05:37

本文选题：VEGF + FLK1　；参考：《大理学院》2014年硕士论文

【摘要】：目的观察VEGF及受体FLK1在大鼠肝缺血再灌注后肝再生中的表达及分布，探讨大鼠肝再生过程中的分子调控机制。方法健康成年SD大鼠40只，随机分为两组，模型组35只，正常对照组5只，用10%的水合氯醛以0.3~0.35ml/100g的剂量进行腹腔麻醉，模型组采用三联阻断法阻断肝蒂20min后行肝大部（约70%）切除。正常对照组麻醉后打开腹腔未阻断肝血流，处死后直接取正常肝组织。模型组分别在术后3h，6h，24h，2d，3d，5d，7d后处死大鼠，切取残肝称湿重后留取1×1×1cm肝组织在10%的中性福尔马林溶液内进行固定24h-48h，然后分别给予脱水、透明及浸蜡等组织学处理，采用石蜡包埋后进行连续切片，H-E染色观察肝组织形态变化，免疫组化PV-9001超敏两步法检测正常肝组织及模型组各个时间点VEGF及受体FLK1在肝再生中的表达及变化。结果与正常对照组相比，模型组肝组织内可见较多炎性细胞，肝细胞索之间及门管区的内皮细胞充血水肿，肝血窦明显增宽，肝血窦中白细胞增多，主要为中性粒细胞。VEGF在正常肝组织中表达较弱，阳性细胞主要为门管区血管内皮细胞，数量较少，周围的肝细胞未见表达。模型组术后3h-6h，VEGF表达逐渐增强，术后1d时VEGF阳性细胞显著增多，主要分布在门管区周围；3d时VEGF阳性反应达到高峰，整个肝小叶的肝细胞均呈阳性反应，并且越靠近中央静脉阳性反应越强；术后5d，VEGF表达开始下降，7d时基本恢复至正常对照组水平。VEGF受体FLK1在正常肝组织中呈阴性反应。模型组术后3h，肝细胞开始出现FLK1弱阳性反应，，阳性反应主要分布于肝细胞膜及细胞浆，24h时表达达到高峰，术后2d时开始逐渐下降，7d时基本恢复正常水平。结论肝再生过程中VEGF与受体FLK1的表达及分布存在着时间和空间差异性，VEGF与其受体FLK结合后可能通过刺激血管内皮细胞分裂、增殖而诱导新生血管形成，从而促进肝再生。
[Abstract]:Objective to investigate the expression and distribution of VEGF and receptor FLK1 in rat liver regeneration after ischemia-reperfusion. Methods Forty healthy adult SD rats were randomly divided into two groups: the model group (n = 35) and the normal control group (n = 5). 10% chloral hydrate was used to anesthetize the rats intraperitoneally with the dose of 0.3~0.35ml/100g. In the model group, transhepatic resection was performed after 20min was blocked by triple occlusion. The normal control group did not block the hepatic blood flow after anesthesia, and took the normal liver tissue directly after death. The rats in the model group were killed at 3 h, 6 h, 24 h, 2 h, 3 d and 5 d after the operation, and 1 脳 1 脳 1cm liver tissue was fixed in 10% neutral formalin solution for 24 h 48 hours, then treated with dehydration, transparency and wax immersion respectively. The morphologic changes of liver tissue were observed by paraffin-embedded serial sections with H-E staining. The expression and changes of VEGF and receptor FLK1 in normal liver tissue and model group were detected by immunohistochemical PV-9001 hypersensitivity two-step method. Results compared with the normal control group, more inflammatory cells were found in the liver tissue of the model group. The endothelial cells between the hepatic cord and the portal area were congested and edema, the sinusoids of the liver were obviously enlarged, and the white blood cells in the sinusoidal of the liver were increased. The expression of neutrophil. VEGF was weak in normal liver tissue and the positive cells were mainly vascular endothelial cells in portal area. In the model group, the expression of VEGF increased gradually at 3h-6 h postoperatively, and increased significantly on the 1st day after operation. The positive expression of VEGF reached its peak at 3 days around the hilar area, and the hepatocytes of the whole hepatic lobule showed positive reaction. The expression of VEGF-1 returned to the level of normal control group after 5 days, and the expression of VEGFR FLK1 was negative in normal liver tissue. In the model group, there was a weak positive reaction of FLK1 in hepatocytes at 3 h after operation, the positive reaction was mainly distributed in the liver cell membrane and cytoplasm at 24 h and reached the peak at 24 h, and gradually decreased at 2 d after operation and returned to normal level at 7 d after operation. Conclusion there is a temporal and spatial difference in the expression and distribution of VEGF and receptor FLK1 during liver regeneration. After VEGF binds to its receptor FLK, it may induce angiogenesis by stimulating vascular endothelial cell division and proliferation, thus promoting liver regeneration.
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R657.3

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