LPS对人类肠道微血管内皮细胞水通道蛋白-1表达及其功能的影响
发布时间：2019-06-02 15:03
【摘要】：目的通过观察LPS对人类肠粘膜微血管内皮细胞水通道蛋白-1表达及m-RNA水平的影响,探讨水通道蛋白在脓毒症患者肠微血管内皮细胞通透性改变中的意义。 方法在体外培养的人类肠粘膜微血管内皮细胞(]HIMECs),根据LPS的浓度分为4组：低浓度组(LPS中浓度组(LPS1mg/L)、高浓度组(LPS10mg/L)、对照组只加培养基。与之培养6h、12h、24h后,应用细胞免疫荧光及RT-PCR法测定AQP1表达及其m-RNA的水平。 结果LPS作用于肠微血管内皮细胞,6h后AQP-1蛋白及其m-RNA的表达开始下降,其中免疫荧光显示LPS刺激组AQP1蛋白表达量逐渐降低,与对照组比较有显著性差异(p0.01),并且呈现一定的浓度及时间相关性趋势。RT-PCR显示LPS刺激组AQP1-mRNA表达量逐步降低,与对照组比较有显著性差异(p0.05),并且呈现一定的浓度及时间相关性趋势。 结论LPS刺激HIMECs后,AQP1出现蛋白表达量下降及其m-RNA水平降低,表明AQP1可能在急性肠损伤后的肠壁水肿、肠道通透性增加有关,为进一步实验研究提供理论及技术基础。
[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.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7
本文编号：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.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7
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