炎症缺血缺氧损伤对体外培养的大脑皮层星形胶质细胞中内皮性脂酶表达的影响

Effect of ischemia /hypoxia injury in inflammation condition on expression of endothelial lipase in cultural cortical astrocyte in vitro

目的:研究星形胶质细胞在炎症状态下缺血损伤对内皮性脂酶(endothelial lipase,EL)表达及活性的影响。方法:实验分为正常对照组、单纯损伤组和炎症损伤组。取新生大鼠脑皮质,体外分离培养纯化星形胶质细胞;单纯损伤组为对星形胶质细胞进行缺血缺氧损伤,炎症损伤组为白细胞共培养下的星形胶质细胞进行缺血缺氧损伤;两组损伤后分别在0,1,3,6,12,24,48 h不同时间点用逆转录PCR、实时荧光定量PCR和Western Blot检测EL的表达变化。结果:缺血损伤的星形胶质细胞中EL的表达比正常对照组偏高,并随着损伤时间的延长逐渐增高,在损伤后6 h和12 h达到峰值、随后下降。当加入炎症细胞后,炎症反应进一步促进EL在缺血缺氧损伤星形胶质细胞中的表达增加。结论:缺血缺氧损伤所介导的星形胶质细胞内EL的表达变化以及对该过程的促进作用可能直接影响到星形胶质细胞脂质的再利用,对损伤神经元的修复有极其重要的参考价值和临床意义。

Objective： To study the effect of ischemia/hypoxia injury in inflammation condition on expression of endo-thelial lipase（ EL）in astrocytes. Methods： The experiments were divided into normal control group, injury group and combination of inflammation and injury group. Purified cortical astroeytes were prepared from neonatal rats. Injury group was ischemia/hypoxia injured astroeyte and inflammation injury group was ischemia/hypoxia injured astrocyte after being co-culture with leukoeytes. The different expression of EL between injury group and inflammation injury group were at 0, 1,3,6,12,24,48 h examined by semi-quantitative reverse-transcriptional PCR and quantitative real-time PCR and Western Blot assay. Results： The expression of EL in ischemia/hypoxia injured astrocyte was relatively higher than that in normal control group, and gradually increased with the extension of time, and reached a peak at 6 h and 12 h after injury, then declined. After treatment of inflammatory cells, inflammatory response further promoted EL expression increase in hypoxic-ischemie injured astroeytes. Conclusion： The variability expression of EL in astrocyte induced by ischemia/hypoxia injury and its promotion could directly affect the lipid reuse of astrocytes, which have an extremely important reference value and clinical significance to the repair of damaged neurons.