本文選題：VEGF + FLK1��； 參考：《大理學(xué)院》2014年碩士論文

【摘要】：目的觀察VEGF及受體FLK1在大鼠肝缺血再灌注后肝再生中的表達(dá)及分布，探討大鼠肝再生過(guò)程中的分子調(diào)控機(jī)制。 方法健康成年SD大鼠40只，隨機(jī)分為兩組，模型組35只，正常對(duì)照組5只，用10%的水合氯醛以0.3~0.35ml/100g的劑量進(jìn)行腹腔麻醉，模型組采用三聯(lián)阻斷法阻斷肝蒂20min后行肝大部（約70%）切除。正常對(duì)照組麻醉后打開(kāi)腹腔未阻斷肝血流，處死后直接取正常肝組織。模型組分別在術(shù)后3h，6h，24h，2d，3d，5d，7d后處死大鼠，切取殘肝稱濕重后留取1×1×1cm肝組織在10%的中性福爾馬林溶液內(nèi)進(jìn)行固定24h-48h，然后分別給予脫水、透明及浸蠟等組織學(xué)處理，采用石蠟包埋后進(jìn)行連續(xù)切片，H-E染色觀察肝組織形態(tài)變化，免疫組化PV-9001超敏兩步法檢測(cè)正常肝組織及模型組各個(gè)時(shí)間點(diǎn)VEGF及受體FLK1在肝再生中的表達(dá)及變化。 結(jié)果與正常對(duì)照組相比，模型組肝組織內(nèi)可見(jiàn)較多炎性細(xì)胞，肝細(xì)胞索之間及門(mén)管區(qū)的內(nèi)皮細(xì)胞充血水腫，肝血竇明顯增寬，肝血竇中白細(xì)胞增多，主要為中性粒細(xì)胞。VEGF在正常肝組織中表達(dá)較弱，陽(yáng)性細(xì)胞主要為門(mén)管區(qū)血管內(nèi)皮細(xì)胞，數(shù)量較少，周?chē)母渭?xì)胞未見(jiàn)表達(dá)。模型組術(shù)后3h-6h，VEGF表達(dá)逐漸增強(qiáng)，術(shù)后1d時(shí)VEGF陽(yáng)性細(xì)胞顯著增多，主要分布在門(mén)管區(qū)周?chē)?d時(shí)VEGF陽(yáng)性反應(yīng)達(dá)到高峰，整個(gè)肝小葉的肝細(xì)胞均呈陽(yáng)性反應(yīng)，并且越靠近中央靜脈陽(yáng)性反應(yīng)越強(qiáng)；術(shù)后5d，VEGF表達(dá)開(kāi)始下降，7d時(shí)基本恢復(fù)至正常對(duì)照組水平。VEGF受體FLK1在正常肝組織中呈陰性反應(yīng)。模型組術(shù)后3h，肝細(xì)胞開(kāi)始出現(xiàn)FLK1弱陽(yáng)性反應(yīng)，，陽(yáng)性反應(yīng)主要分布于肝細(xì)胞膜及細(xì)胞漿，24h時(shí)表達(dá)達(dá)到高峰，術(shù)后2d時(shí)開(kāi)始逐漸下降，7d時(shí)基本恢復(fù)正常水平。 結(jié)論肝再生過(guò)程中VEGF與受體FLK1的表達(dá)及分布存在著時(shí)間和空間差異性，VEGF與其受體FLK結(jié)合后可能通過(guò)刺激血管內(nèi)皮細(xì)胞分裂、增殖而誘導(dǎo)新生血管形成，從而促進(jìn)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.
【學(xué)位授予單位】：大理學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R657.3

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