[目的]丙泊酚改善重症脓毒症小鼠脑保护作用的研究。[方法]将48只雄性小鼠随机分为对照组、LPS组、PF组、PF+ML385组,每组12只。通过苏木素-伊红染色评估各组小鼠海马组织的病理变化;通过原位末端标记法分析海马组织中神经元的凋亡率;...[目的]丙泊酚改善重症脓毒症小鼠脑保护作用的研究。[方法]将48只雄性小鼠随机分为对照组、LPS组、PF组、PF+ML385组,每组12只。通过苏木素-伊红染色评估各组小鼠海马组织的病理变化;通过原位末端标记法分析海马组织中神经元的凋亡率;通过酶联免疫吸附实验分析血清中TNF-α、IL-6、SOD和CAT的水平;通过蛋白免疫印迹实验分析海马组织中TLR4/MD-2的蛋白表达水平。[结果]与LPS组比较,PF组与PF+ML385组小鼠海马组织区域炎症细胞聚集减少,神经元损伤减轻,神经元凋亡率降低,血清炎症因子(TNF-α和IL-6)水平降低,血清抗氧化酶(SOD和CAT)活性水平增加,海马组织的TLR4、MD-2蛋白表达水平降低。与PF组比较,PF+ML385组小鼠海马组织神经元凋亡率降低(13.26%±2.19%vs 9.28%±1.58%;P<0.05),血清炎症因子(TNF-α、IL-6)水平降低(15.61±1.29 vs 10.28±1.46 pg/mL;30.28±1.11 vs 23.56±2.13 pg/mL;P<0.05),血清抗氧化酶(SOD、CAT)活性水平增加(13.69±1.72 vs 19.23±2.39 U/mg protein;12.97±2.26 vs 17.38±1.47 U/mg protein;P<0.05),海马组织的TLR4、MD-2蛋白表达水平降低(0.65±0.03 vs 0.33±0.09;0.72±0.08 vs 0.41±0.04;P<0.05)。[结论]丙泊酚能够改善脓毒症小鼠海马组织的神经元损伤,减少神经元凋亡率和炎症因子水平,增加抗氧化酶水平。丙泊酚的这一作用与抑制TLR4/MD-2通路密切相关。展开更多
We isolated cDNA encoding porcine MyD88 （poMyD88） from Peyer＇s patches （Pps） of GALT. The complete open reading frame （ORF） of poMyD88 contains 879 bp encoding a deduced 293 aa residues. The amino acid sequence...We isolated cDNA encoding porcine MyD88 （poMyD88） from Peyer＇s patches （Pps） of GALT. The complete open reading frame （ORF） of poMyD88 contains 879 bp encoding a deduced 293 aa residues. The amino acid sequence of poMyD88 was characterized by N-terminal death, intermediate and C-terminal Toll/IL-1 receptor （TIR） domains. The putative poMyD88 protein shares a higher level of homology with its human （87.2% amino acid identity） than with its mouse （77.4% amino acid identity） counterpart. Overexpression of poMyD88 participated in the further enhanced activation of NF-w.B in human embryonic kidney （HEK） 293 cells expressing porcine TLR2 and porcine TLR4/MD-2, but not porcine RP105/MD-1 after stimulation with the corresponding ligands. The expression levels of MyD88 were highest in the spleen and mesenteric lymph nodes （MLNs）, and lower in digestive tissues of newborn swine. In adult swine, the expression levels in the digestive tissues were lower than those in MLNs and the spleen. These results suggest that an MyD88-dependent signaling pathway is present in newborn as well as in adult swine and that it is involved in the innate immune system of these animals.展开更多
文摘[目的]丙泊酚改善重症脓毒症小鼠脑保护作用的研究。[方法]将48只雄性小鼠随机分为对照组、LPS组、PF组、PF+ML385组,每组12只。通过苏木素-伊红染色评估各组小鼠海马组织的病理变化;通过原位末端标记法分析海马组织中神经元的凋亡率;通过酶联免疫吸附实验分析血清中TNF-α、IL-6、SOD和CAT的水平;通过蛋白免疫印迹实验分析海马组织中TLR4/MD-2的蛋白表达水平。[结果]与LPS组比较,PF组与PF+ML385组小鼠海马组织区域炎症细胞聚集减少,神经元损伤减轻,神经元凋亡率降低,血清炎症因子(TNF-α和IL-6)水平降低,血清抗氧化酶(SOD和CAT)活性水平增加,海马组织的TLR4、MD-2蛋白表达水平降低。与PF组比较,PF+ML385组小鼠海马组织神经元凋亡率降低(13.26%±2.19%vs 9.28%±1.58%;P<0.05),血清炎症因子(TNF-α、IL-6)水平降低(15.61±1.29 vs 10.28±1.46 pg/mL;30.28±1.11 vs 23.56±2.13 pg/mL;P<0.05),血清抗氧化酶(SOD、CAT)活性水平增加(13.69±1.72 vs 19.23±2.39 U/mg protein;12.97±2.26 vs 17.38±1.47 U/mg protein;P<0.05),海马组织的TLR4、MD-2蛋白表达水平降低(0.65±0.03 vs 0.33±0.09;0.72±0.08 vs 0.41±0.04;P<0.05)。[结论]丙泊酚能够改善脓毒症小鼠海马组织的神经元损伤,减少神经元凋亡率和炎症因子水平,增加抗氧化酶水平。丙泊酚的这一作用与抑制TLR4/MD-2通路密切相关。
文摘目的探讨脂多糖作用小鼠RAW264.7巨噬细胞24 h内细胞表面TLR4/CD14/MD-2受体复合物表达的变化特点。方法分别用低剂量(100 ng/ml)和高剂量(1 000 ng/ml)脂多糖刺激RAW264.7,应用RT-PCR检测TLR4、CD14、MD-2m RNA水平的变化,ELISA检测细胞上清TNF-α含量的变化。结果低剂量脂多糖下调TLR4 m RNA表达,14 h降至最低,24 h维持在低水平表达;CD14和MD-2 m RNA表达增加。高剂量脂多糖上调TLR4 m RNA表达,1 h上升至高峰,此后维持在高水平表达;CD14和MD-2 m RNA表达也明显增强。高剂量脂多糖作用后TNF-α含量显著高于低剂量脂多糖(P<0.05)。结论高剂量脂多糖逆转低剂量脂多糖下调的TLR4表达,提高TLR4/CD14/MD-2受体复合物的表达水平,放大下游信号转导通路的炎症效应。
文摘We isolated cDNA encoding porcine MyD88 （poMyD88） from Peyer＇s patches （Pps） of GALT. The complete open reading frame （ORF） of poMyD88 contains 879 bp encoding a deduced 293 aa residues. The amino acid sequence of poMyD88 was characterized by N-terminal death, intermediate and C-terminal Toll/IL-1 receptor （TIR） domains. The putative poMyD88 protein shares a higher level of homology with its human （87.2% amino acid identity） than with its mouse （77.4% amino acid identity） counterpart. Overexpression of poMyD88 participated in the further enhanced activation of NF-w.B in human embryonic kidney （HEK） 293 cells expressing porcine TLR2 and porcine TLR4/MD-2, but not porcine RP105/MD-1 after stimulation with the corresponding ligands. The expression levels of MyD88 were highest in the spleen and mesenteric lymph nodes （MLNs）, and lower in digestive tissues of newborn swine. In adult swine, the expression levels in the digestive tissues were lower than those in MLNs and the spleen. These results suggest that an MyD88-dependent signaling pathway is present in newborn as well as in adult swine and that it is involved in the innate immune system of these animals.
