本文選題：HIF-1α　切入點(diǎn)：VEGF　出處：《蘇州大學(xué)》2006年碩士論文

【摘要】： 目的:從小鼠體內(nèi)和體外兩方面,探討LPS誘導(dǎo)的急性炎癥組織中和體外培養(yǎng)的巨噬細(xì)胞內(nèi)HIF-1α表達(dá)的改變及其意義 方法:將Balb/c小鼠分為正常對照組及炎癥組,炎癥組采用卡介苗(BCG)配合脂多糖(LPS)尾靜脈注射法建立小鼠急性全身性炎癥損傷模型。取出肝臟和肺臟進(jìn)行常規(guī)石蠟切片、HE染色觀察臟器的病理改變,免疫組織化學(xué)SP法觀察HIF-1α及VEGF在組織中的表達(dá)改變。另取正常Balb/c小鼠,分離腹腔巨噬細(xì)胞分為正常對照組、LPS刺激組,刺激組給予LPS、對照組給予PBS作用10小時(shí)后,采用RT-PCR法半定量檢測HIF-1α、VEGF基因mRNA的表達(dá)水平,應(yīng)用細(xì)胞免疫化學(xué)染色法檢測巨噬細(xì)胞HIF-1α蛋白的表達(dá)水平。 結(jié)果:炎癥組小鼠肝臟及肺組織HIF-1α、VEGF表達(dá)較正常對照組顯著增加(p均0.01)。體外培養(yǎng)中,與正常對照組比較,LPS刺激的巨噬細(xì)胞HIF-1α基因表達(dá)無明顯改變(P0.05),但HIF-1α蛋白表達(dá)顯著增加且VEGF基因表達(dá)上調(diào)(p均0.01)。 結(jié)論:由LPS引起小鼠全身急性炎性時(shí),肝和肺組織表達(dá)的HIF-1α及VEGF蛋白水平升高;體外常氧條件下,LPS刺激的小鼠巨噬細(xì)胞可在轉(zhuǎn)錄后水平上調(diào)HIF-1α蛋白質(zhì)的表達(dá),并可上調(diào)靶基因VEGF的表達(dá);提示常氧條件下,LPS可通過轉(zhuǎn)錄后水平上調(diào)HIF-1α蛋白的表達(dá)從而參與體內(nèi)的急性炎癥反應(yīng)。
[Abstract]:Objective: to investigate the changes and significance of HIF-1 偽 expression in acute inflammatory tissue induced by LPS and macrophages cultured in vitro and in vivo in mice. Methods: Balb/c mice were divided into normal control group and inflammatory group. The acute systemic inflammatory injury model of mice was established by BCG combined with lipopolysaccharide (LPS) injection into tail vein. The liver and lung were taken out for routine paraffin section and HE staining to observe the pathological changes of viscera. Immunohistochemical SP method was used to observe the expression of HIF-1 偽 and VEGF in normal Balb/c mice. The peritoneal macrophages were isolated and divided into two groups: the normal control group was stimulated by PBS, and the control group was treated with PBS for 10 hours. The expression of HIF-1 偽 -VEGF gene mRNA was detected by semi-quantitative RT-PCR method, and the expression of HIF-1 偽 protein in macrophages was detected by cytoimmunochemical staining. Results: compared with the normal control group, the expression of HIF-1 偽 -VEGF in the liver and lung tissues of the inflammatory group increased significantly. The expression of HIF-1 偽 gene in LPS-stimulated macrophages was not significantly changed, but the expression of HIF-1 偽 protein was significantly increased and the expression of VEGF gene was up-regulated. Conclusion: the expression of HIF-1 偽 and VEGF protein in liver and lung tissues increased during acute inflammation induced by LPS, and the expression of HIF-1 偽 protein was up-regulated by LPS-stimulated mouse macrophages at posttranscriptional level. The expression of target gene VEGF could be upregulated, which suggested that lipopolysaccharide could up-regulate the expression of HIF-1 偽 protein at posttranscriptional level and participate in the acute inflammatory response in vivo.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R363

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