目的:探究锌离子对大鼠肺缺血/再灌注损伤(lung ischemia/reperfusion injury,LIRI)的保护作用与机制。方法:将32只6~8周的SPF级SD大鼠,以每组8只随机分为4组:对照(control)组、缺血/再灌注组(ischemia/reperfusion,I/R)、氯化锌+I/R(Zn...目的:探究锌离子对大鼠肺缺血/再灌注损伤(lung ischemia/reperfusion injury,LIRI)的保护作用与机制。方法:将32只6~8周的SPF级SD大鼠,以每组8只随机分为4组:对照(control)组、缺血/再灌注组(ischemia/reperfusion,I/R)、氯化锌+I/R(ZnCl_(2)+I/R)组和PI3K抑制剂LY294002+ZnCl_(2)+I/R组。电感耦合等离子体质谱(induc⁃tively coupled plasma mass spectrometry,ICP-MS)检测各组肺组织锌离子浓度;HE染色、肺泡损伤指数、肺湿干重比检测各组肺组织损伤程度;qPCR法检测肺组织中溶质载体家族39成员8(solute carrier family 39 member 8,SLC39A8/ZIP8)的表达水平;TUNEL检测各组肺组织凋亡水平;Western blot检测各组肺组织caspase-3表达水平、PI3K/AKT/糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK-3β)磷酸化水平,以及ZIP8和溶质载体家族30成员9(solute carrier family 30 member 9,SLC30A9/ZNT9)表达水平;线粒体提取试剂盒及JC-1线粒体膜电位检测试剂盒测定线粒体膜电位变化。结果:与I/R组相比,ZnCl_(2)+I/R组锌离子水平上升(P<0.01),qPCR结果显示ZIP8表达水平上升(P<0.01),Western blot结果表明PI3K/AKT/GSK-3β磷酸化水平上升(P<0.01或P<0.05),cleaved caspase-3/pro caspase-3水平下降(P<0.01),ZIP8水平上升(P<0.05),ZNT9水平无显著差异(P>0.05),线粒体膜电位上升(P<0.01),TUNEL结果显示凋亡水平减弱(P<0.01),HE染色、总肺含水量(total lung water content,TLW)、肺组织定量评价指标(lung index of quantitative assessment of histology,IQA)及肺组织湿干重比结果显示损伤显著减轻(P<0.05或P<0.01)。与ZnCl_(2)+I/R组相比,LY294002+ZnCl_(2)+I/R组锌离子水平下降(P<0.05),qPCR结果显示ZIP8表达水平下降(P<0.01),Western blot结果表明PI3K/AKT/GSK-3β磷酸化水平下降(P<0.01),cleaved caspase-3/procas⁃pase-3水平上升(P<0.01),ZIP8水平下降(P<0.05),ZNT9水平无显著差异(P>0.05),线粒体膜电位下降(P<0.01),TUNEL结果显示凋亡水平增强(P<0.05),HE染色、TLW、IQA%及肺组织湿干重比结果显示损伤显著加重(P<0.05或P<0.01)。结论:氯化锌可通过激活大鼠PI3K/AKT通路,导致GSK-3β失活,稳定线粒体膜电位水平,抑制细胞凋亡,从而发挥对缺血/再灌注损伤肺的保护作用。展开更多
A method for simultaneous determination of nine cations including Fe 3+, Cu 2+, Ni 2+, Zn 2+, Co 2+, Fe 2+, Cd 2+, Mg 2+ and Ca 2+ by single-column ion chromatography with a conductivity detector was developed. The op...A method for simultaneous determination of nine cations including Fe 3+, Cu 2+, Ni 2+, Zn 2+, Co 2+, Fe 2+, Cd 2+, Mg 2+ and Ca 2+ by single-column ion chromatography with a conductivity detector was developed. The optimum composition of eluent was 6 mmol/L malic acid and 3 mmol/L ethylenediamine(pH=4 0). The effects of malic acid concentration, ethylenediamine concentration and pH value on retention time were studied in details. The method was used to determine Fe, Cu, Zn, Mg and Ca in whole blood and serum with satisfactory results.展开更多
文摘目的:探究锌离子对大鼠肺缺血/再灌注损伤(lung ischemia/reperfusion injury,LIRI)的保护作用与机制。方法:将32只6~8周的SPF级SD大鼠,以每组8只随机分为4组:对照(control)组、缺血/再灌注组(ischemia/reperfusion,I/R)、氯化锌+I/R(ZnCl_(2)+I/R)组和PI3K抑制剂LY294002+ZnCl_(2)+I/R组。电感耦合等离子体质谱(induc⁃tively coupled plasma mass spectrometry,ICP-MS)检测各组肺组织锌离子浓度;HE染色、肺泡损伤指数、肺湿干重比检测各组肺组织损伤程度;qPCR法检测肺组织中溶质载体家族39成员8(solute carrier family 39 member 8,SLC39A8/ZIP8)的表达水平;TUNEL检测各组肺组织凋亡水平;Western blot检测各组肺组织caspase-3表达水平、PI3K/AKT/糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK-3β)磷酸化水平,以及ZIP8和溶质载体家族30成员9(solute carrier family 30 member 9,SLC30A9/ZNT9)表达水平;线粒体提取试剂盒及JC-1线粒体膜电位检测试剂盒测定线粒体膜电位变化。结果:与I/R组相比,ZnCl_(2)+I/R组锌离子水平上升(P<0.01),qPCR结果显示ZIP8表达水平上升(P<0.01),Western blot结果表明PI3K/AKT/GSK-3β磷酸化水平上升(P<0.01或P<0.05),cleaved caspase-3/pro caspase-3水平下降(P<0.01),ZIP8水平上升(P<0.05),ZNT9水平无显著差异(P>0.05),线粒体膜电位上升(P<0.01),TUNEL结果显示凋亡水平减弱(P<0.01),HE染色、总肺含水量(total lung water content,TLW)、肺组织定量评价指标(lung index of quantitative assessment of histology,IQA)及肺组织湿干重比结果显示损伤显著减轻(P<0.05或P<0.01)。与ZnCl_(2)+I/R组相比,LY294002+ZnCl_(2)+I/R组锌离子水平下降(P<0.05),qPCR结果显示ZIP8表达水平下降(P<0.01),Western blot结果表明PI3K/AKT/GSK-3β磷酸化水平下降(P<0.01),cleaved caspase-3/procas⁃pase-3水平上升(P<0.01),ZIP8水平下降(P<0.05),ZNT9水平无显著差异(P>0.05),线粒体膜电位下降(P<0.01),TUNEL结果显示凋亡水平增强(P<0.05),HE染色、TLW、IQA%及肺组织湿干重比结果显示损伤显著加重(P<0.05或P<0.01)。结论:氯化锌可通过激活大鼠PI3K/AKT通路,导致GSK-3β失活,稳定线粒体膜电位水平,抑制细胞凋亡,从而发挥对缺血/再灌注损伤肺的保护作用。
文摘A method for simultaneous determination of nine cations including Fe 3+, Cu 2+, Ni 2+, Zn 2+, Co 2+, Fe 2+, Cd 2+, Mg 2+ and Ca 2+ by single-column ion chromatography with a conductivity detector was developed. The optimum composition of eluent was 6 mmol/L malic acid and 3 mmol/L ethylenediamine(pH=4 0). The effects of malic acid concentration, ethylenediamine concentration and pH value on retention time were studied in details. The method was used to determine Fe, Cu, Zn, Mg and Ca in whole blood and serum with satisfactory results.
