LPS對人類腸道微血管內皮細胞水通道蛋白-1表達及其功能的影響
發(fā)布時間：2019-06-02 15:03
【摘要】：目的通過觀察LPS對人類腸粘膜微血管內皮細胞水通道蛋白-1表達及m-RNA水平的影響,探討水通道蛋白在膿毒癥患者腸微血管內皮細胞通透性改變中的意義。 方法在體外培養(yǎng)的人類腸粘膜微血管內皮細胞(]HIMECs),根據LPS的濃度分為4組：低濃度組(LPS中濃度組(LPS1mg/L)、高濃度組(LPS10mg/L)、對照組只加培養(yǎng)基。與之培養(yǎng)6h、12h、24h后,應用細胞免疫熒光及RT-PCR法測定AQP1表達及其m-RNA的水平。 結果LPS作用于腸微血管內皮細胞,6h后AQP-1蛋白及其m-RNA的表達開始下降,其中免疫熒光顯示LPS刺激組AQP1蛋白表達量逐漸降低,與對照組比較有顯著性差異(p0.01),并且呈現一定的濃度及時間相關性趨勢。RT-PCR顯示LPS刺激組AQP1-mRNA表達量逐步降低,與對照組比較有顯著性差異(p0.05),并且呈現一定的濃度及時間相關性趨勢。 結論LPS刺激HIMECs后,AQP1出現蛋白表達量下降及其m-RNA水平降低,表明AQP1可能在急性腸損傷后的腸壁水腫、腸道通透性增加有關,為進一步實驗研究提供理論及技術基礎。
[Abstract]:Objective to observe the effect of LPS on the expression of aquaporin-1 and the level of m-RNA in human intestinal microvascular endothelial cells, and to explore the significance of aquaporin in the change of intestinal microvascular endothelial cell permeability in sepsis patients. Methods Human intestinal microvascular endothelial cells (HIMECs),) cultured in vitro were divided into four groups according to the concentration of LPS: low concentration group (LPS medium concentration group (LPS1mg/L), high concentration group (LPS10mg/L), control group only supplemented with culture medium). After 6 h, 12 h and 24 h, the expression of AQP1 and the level of m-RNA were detected by immunofluorescence and RT-PCR assay. Results the expression of AQP-1 protein and its m-RNA in intestinal microvascular endothelial cells treated with LPS began to decrease after 6 hours. Immunofluorescence showed that the expression of AQP1 protein in LPS stimulated group decreased gradually, which was significantly different from that in the control group (p0.01). RT-PCR showed that the expression of AQP1-mRNA in LPS stimulation group decreased gradually, which was significantly different from that in control group (p0.05), and showed a certain concentration and time correlation trend. Conclusion after LPS stimulation of HIMECs, the protein expression and m-RNA level of AQP1 decreased, indicating that AQP1 may be related to intestinal wall edema and intestinal permeability after acute intestinal injury, which provides a theoretical and technical basis for further experimental study.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R459.7
本文編號：2491199
[Abstract]:Objective to observe the effect of LPS on the expression of aquaporin-1 and the level of m-RNA in human intestinal microvascular endothelial cells, and to explore the significance of aquaporin in the change of intestinal microvascular endothelial cell permeability in sepsis patients. Methods Human intestinal microvascular endothelial cells (HIMECs),) cultured in vitro were divided into four groups according to the concentration of LPS: low concentration group (LPS medium concentration group (LPS1mg/L), high concentration group (LPS10mg/L), control group only supplemented with culture medium). After 6 h, 12 h and 24 h, the expression of AQP1 and the level of m-RNA were detected by immunofluorescence and RT-PCR assay. Results the expression of AQP-1 protein and its m-RNA in intestinal microvascular endothelial cells treated with LPS began to decrease after 6 hours. Immunofluorescence showed that the expression of AQP1 protein in LPS stimulated group decreased gradually, which was significantly different from that in the control group (p0.01). RT-PCR showed that the expression of AQP1-mRNA in LPS stimulation group decreased gradually, which was significantly different from that in control group (p0.05), and showed a certain concentration and time correlation trend. Conclusion after LPS stimulation of HIMECs, the protein expression and m-RNA level of AQP1 decreased, indicating that AQP1 may be related to intestinal wall edema and intestinal permeability after acute intestinal injury, which provides a theoretical and technical basis for further experimental study.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R459.7
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