摘要
目的:探究肠上皮细胞炎症发生过程中细胞能量代谢损伤对肠上皮细胞线粒体结构和功能的影响。方法:采用过氧化氢构建肠上皮细胞体外炎症模型,给予0μmol/L和100μmol/L脱氧胆酸(DCA)干预5 d,分别记为模型组和实验组,同时以不做任何处理的人正常结肠上皮细胞NCM460为空白对照组,采用透射电镜观察各组肠上皮细胞线粒体超微结构差异;采用CCK8实验检测各组细胞活力;采用ATP试剂盒检测各组细胞ATP生成量;采用激光共聚焦显微镜检测细胞线粒体膜电位改变;检测各组肠上皮细胞耗氧率、呼吸代谢指标及活性氧百分含量差异情况。结果:空白对照组线粒体形态结构正常;模型组线粒体肿胀,内嵴紊乱,甚至断裂消失;实验组线粒体出现气球样自噬泡。与空白对照组比较,模型组和实验组肠上皮细胞活性受到明显抑制(P<0.05),细胞线粒体ATP生成量及ATP酶活性明显较低(P<0.05),线粒体膜电位也明显较低(P<0.05),细胞耗氧率及基础呼吸、质子渗漏、最大呼吸、储备呼吸率、非线粒体呼吸等呼吸指标也明显较低(P<0.05),细胞活性氧百分含量则明显较高(P<0.05);与模型组比较,实验组肠上皮细胞活性抑制更明显(P<0.05),细胞线粒体ATP生成量、ATP酶活性、线粒体膜电位均进一步降低(P<0.05),细胞耗氧率及基础呼吸、质子渗漏、最大呼吸、储备呼吸率、非线粒体呼吸等呼吸指标进一步减少(P<0.05),细胞活性氧百分含量则进一步增高(P<0.05)。结论:线粒体参与肠上皮细胞炎症的能量代谢调节,其作用与线粒体呼吸功能抑制及膜电位下降有关。
Objective:To investigate the effects of cellular energy metabolism damage on the mitochondrial structure and function of intestinal epithelial cells during the inflammatory process.Methods:The hydrogen peroxide(H2O2)was used to construct intestinal epithelial cells in vitro model of inflammation,0μmol/L and 100μmol/L deoxycholic acid(DCA)intervention in 5 d,respectively for the model group and experimental group,at the same time with those who do not do any processing normal colon epithelial cells NCM460 as blank control group,with transmission electron microscope each enterocyte mitochondrial ultrastructure of difference;the cell activity of each group was measured by CCK8 experiment.ATP production of each group was detected by ATP kit.The changes of mitochondrial membrane potential were detected by laser confocal microscopy.The differences of oxygen consumption rate,respiratory metabolism index and reactive oxygen content in intestinal epithelial cells were detected.Results:In the blank control group,mitochondria were normal in morphology and structure;in the model group,mitochondria were swollen,internal cristae were disordered,and even rupture disappeared;in the experimental group,balloon-like autophagy vesicles appeared.Compared with blank control group,model group and experimental group of intestinal epithelial cell activity was significantly inhibited(P<0.05),the cells mitochondrial ATP generation and atpase activity significantly lower(P<0.05),mitochondrial membrane potential is also significantly lower(P<0.05),the cell oxygen consumption rate and respiration,proton leakage,maximum,reserve breathing rate,the mitochondrial respiratory breathing indexes such as also significantly lower(P<0.05),active oxygen has a significantly higher percentage of cells(P<0.05).Compared with model group,experimental group activity of intestinal epithelial cells more obvious(P<0.05),the cells mitochondrial ATP generation,atpase activity and mitochondrial membrane potential were further reduced(P<0.05),the cell oxygen consu
作者
马志华
阿迪力江·喀日
阿布来提·阿不都哈尔
MA Zhihua;ADILIJIANG·Kari;ABULAITI·Abduhar(Department of Pediatric Acute and Critical Care Medicine,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,830054,China)
出处
《中国中西医结合消化杂志》
CAS
2021年第7期501-506,共6页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
新疆维吾尔自治区自然科学基金项目(No:2019D01C473)。
关键词
炎症性肠病
肠上皮细胞
线粒体
能量代谢
呼吸代谢
膜电位
inflammatory bowel disease
intestinal epithelial cells
mitochondria
energy metabolism
respiratory metabolism
membrane potential
