摘要
目的探讨TNF-αⅠ型受体(TNFR1)对小鼠脑血管内皮细胞(BVEC)内氧自由基产生及锰超氧化物歧化酶(MnSOD)基因表达的影响。方法体外培养TNFR1基因敲除小鼠脑血管内皮细胞(BVEC/RI)和野生型小鼠脑血管内皮细胞,分别给予5ng/mLTNF-α刺激24h,用荧光显微镜观察细胞内氧自由基的产生,荧光定量PCR法及Western-blot法测定MnSOD基因mRNA和蛋白表达。结果给予TNF-α刺激后,细胞内活性氧(ROS)、锰超氧化物歧化酶基因mRNA及蛋白表达仅在BVEC内明显增高,而在BVEC/RI无明显变化。结论TNF-α可能通过作用于脑血管内皮细胞TNFR1增加细胞内氧自由基产生以及MnSOD基因及蛋白表达,从而介导了细胞氧化应激。
Objective To investigate effects of TNF Receptor Ⅰ （TNFR1） on reactive oxygen species（ROS） and the mRNA and protein expression of manganese superoxide dismutase （MnSOD） gene in mouse brain microvessel endothelial cells. Methods Brain microvessel endothelial cell of TNFR1 knocked out mouse（BVEC/ RI） and wild-type mouse brain microvessel endothelial cell（BVEC） in vitro culture were stimulated respectively with 5 ng/mL TNF-α for 24 hours. The level of ROS was investigated by fluorescence microscope and the mRNA and protein expression of MnSOD in two kinds of cells were determined using fluorescence quantitative PCR and western blot. Results Only the level of ROS and MnSOD mRNA and protein expression in the BVEC increased significantly, while no difference in the BVEC/RI after TNF-α treatment. Conclusions TNFR1 can mediate oxygen stress through affecting ROS and MnSOD in mouse brain microvessel endothelial cells.
出处
《中国神经免疫学和神经病学杂志》
CAS
2008年第6期446-449,476,共5页
Chinese Journal of Neuroimmunology and Neurology
