發(fā)布時(shí)間：2018-08-11 11:10

【摘要】：目的：觀察高糖培養(yǎng)人臍靜脈內(nèi)皮細(xì)胞連接相關(guān)蛋白(occludin、VE-cadherin和connexin40)的表達(dá)變化,探討Ang-1對(duì)高糖環(huán)境下這些內(nèi)皮細(xì)胞連接相關(guān)蛋白的影響。 方法：將體外培養(yǎng)的人臍靜脈內(nèi)皮細(xì)胞隨機(jī)分為正常組(5.5 mmol/L葡萄糖),甘露醇組(5.5 mmol/L葡萄糖+24.5 mmol/L甘露醇),高糖組(30 mmol/L葡萄糖),Ang-1干預(yù)組(30mmol/L葡萄糖+0.25mg/L Ang-1),用相應(yīng)的培養(yǎng)基培養(yǎng)24h、48h后收集細(xì)胞,分別用實(shí)時(shí)定量聚合酶鏈反應(yīng)(Real-Time PCR)檢測(cè)各組臍靜脈內(nèi)皮細(xì)胞occludin、VE-cadherin和connexin 40 mRNA的表達(dá),免疫熒光、免疫組化方法檢測(cè)各組臍靜脈內(nèi)皮細(xì)胞occludin、VE-cadherin和connexin 40蛋白的表達(dá)。 結(jié)果：(1)正常組和甘露醇組細(xì)胞均有occludi、VE-cadherin和connexin 40 mRNA的表達(dá),但兩組間無(wú)明顯統(tǒng)計(jì)學(xué)差異(P0.05)；高糖組細(xì)胞occludin、VE-cadherin和connexin 40 mRNA的表達(dá)均較正常組下調(diào)(P0.01),并呈時(shí)間依賴性(24h vs 48h,P0.05)；Ang-1干預(yù)組細(xì)胞occludin、VE-cadherin和connexin 40 mRNA的表達(dá)雖低于正常組(P0.05),卻較高糖組表達(dá)上調(diào)(P0.05),(2)免疫熒光和組化顯示occludin、VE-cadherin和connexin 40主要表達(dá)于胞膜和胞漿,正常組和甘露醇組細(xì)胞均有表達(dá),但兩組間無(wú)明顯統(tǒng)計(jì)學(xué)差異(P0.05)；高糖組細(xì)胞occludin、VE-cadherin和connexin 40蛋白表達(dá)均較正常組下調(diào)(P0.01),并呈時(shí)間依賴性(24h vs 48h,P0.05)；Ang-1干預(yù)組細(xì)胞occludin、VE-cadherin和connexin 40蛋白表達(dá)雖低于正常組(P0.05),卻較高糖組表達(dá)上調(diào)(P0.05),(3)細(xì)胞occludin、VE-cadherin和connexin蛋白表達(dá)水平彼此呈正相關(guān)。 結(jié)論：高糖培養(yǎng)內(nèi)皮細(xì)胞連接相關(guān)蛋白o(hù)ccludin、VE-cadherin和connexin 40表達(dá)呈時(shí)間依賴性下調(diào)；外源性給予Ang-1,可上調(diào)occludin、VE-cadherin和connexin 40的表達(dá),改善高糖對(duì)內(nèi)皮細(xì)胞連接的損傷。
[Abstract]:Aim: to observe the expression of VE-cadherin and connexin40 in human umbilical vein endothelial cells cultured with high glucose, and to investigate the effect of Ang-1 on them. Methods: cultured human umbilical vein endothelial cells were randomly divided into normal group (5.5 mmol/L glucose), mannitol group (5.5 mmol/L glucose 24.5 mmol/L mannitol) and high glucose group (30 mmol/L glucose) 30mmol/L glucose 0.25mg/L Ang-1. The cells were collected after culture for 24 hours and 48 hours later. The expression of VE-cadherin and connexin 40 mRNA in umbilical vein endothelial cells were detected by real-time quantitative polymerase chain reaction (Real-Time PCR), and the expression of VE-cadherin and connexin 40 protein in endothelial cells of umbilical vein were detected by immunofluorescence and immunohistochemistry. Results: (1) there was no significant difference between the normal group and mannitol group in the expression of connexin 40 mRNA, but there was no significant difference between the two groups (P0.05), the expression of ocdinine VE-cadherin and connexin 40 mRNA in the high glucose group was lower than that in the normal group (P0.01) in a time dependent manner (24 h vs 48 h P 0.05). The expression of VE-cadherin and connexin 40 mRNA in the Ang-1 intervention group was lower than that in the normal group (P0.05), but it was higher than that in the high glucose group (P0.05), (2). The expression of VE-cadherin and connexin 40 were mainly expressed in the cell membrane and cytoplasm in the Ang-1 intervention group, and in the normal group and mannitol group, the expression of VE-cadherin and connexin 40 were found in the cells of the normal group and mannitol group. However, there was no significant difference between the two groups (P0.05), the expression of VE-cadherin and connexin 40 in high glucose group was lower than that in normal group (P0.01), and it was time-dependent (P0.05). The expression of VE-cadherin and connexin 40 in the Ang-1 intervention group was lower than that in the normal group (P0.05), but the expression levels of VE-cadherin and connexin protein were positively correlated with those in the high glucose group (P0.05), (3). Conclusion: the expression of VE-cadherin and connexin 40 in endothelial cells cultured with high glucose was down-regulated in a time-dependent manner, and the expression of VE-cadherin and connexin 40 was up-regulated by exogenous Ang-1.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

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