Establichment of brain oxidative model created by ferric nitrilo-triacetate(Fe-NTA)

去年，Fe-NTA被注射到腹膜液中。今年，为了避免对周围组织的影响，Fe-NTA与脑室内注射（icv）一起给药。采用渗透泵和脑输注试剂盒，将Fe- nta溶液（Fe 0.1mg/ml, 0.25 μl/hr）连续注入侧脑室，观察脑的变化。对照组以同样方式冰注生理盐水。给药2周后，解剖下丘脑、大脑皮层、海马和纹状体，用实时荧光定量PCR检测血红素加氧酶(HO)-1、IL-6和超氧化物歧化酶（SOD） 2 mRNA的表达。HO-1、IL-6 mRNA水平升高，SOD - 2 mRNA水平稳定。Fe-NTA给药4周后，对脑切片进行HE、Pearl染色和TUNEL染色。定量评价采用dot blot法检测4-羟基-2-壬烯醛（4-HNE）， real-time PCR法检测海马和下丘脑HO-1、IL-6、TNFα、SOD 1、SOD 2、SOD 3、Bc12、Bad、FasL mrna。结果HE染色海马明显受损，被炎性组织取代。铁定位于炎症组织和正常组织之间。铁定位诱导神经元细胞在同一区域发生凋亡。海马和下丘脑4-HNE、HO-1、IL-6、IL-1β和tnf - α mRNA显著升高。在对照组中，没有观察到这些变化。冰冻注射Fe-NTA可引起炎症和氧化变化。海马受到明显损伤，可能易受Fe-NTA氧化应激的影响。

In last year, Fe-NTA was injected into peritoneal fluid. In this year, Fe-NTA was administered with intracerebroventricular (icv) injection in order to avoid effects on peripheral tissue. Fe-NTA solusion (Fe 0.1mg/ml, 0.25 μl/hr) was administered continuously into lateral ventricle using osmotic pump and brain infusion kit, and changes of the brains were evaluated. In control, normal saline solution was administered by icy injection in a same manner. After 2 weeks administration, hypothalamus, cerebral cortex, hippocampus, and striatum were dissected, and evaluated in mRNA of hemeoxygenase (HO)-1, IL-6 and superoxidedismutase (SOD) 2 with real-time PCR. Levels of HO-1 and IL-6 mRNA was raised, but SOD 2 mRNA was stable. After 4 weeks administration of Fe-NTA, brain sections were stained in HE, Pearl stain, and TUNEL. In quantitative evaluation, 4-hydroxy-2-nonenal (4-HNE) was measured by dot blot, and mRNAs of HO-1, IL-6, TNFα, SOD 1, SOD 2, SOD 3, Bc12, Bad, and FasL were analyzed by real-time PCR in the hippocampus and hypothalamus. As a result, hippocampus was damaged remarkably and replaced with inflammatory tissue in HE staining. Iron localized between inflammatory tissue and normal tissue. Apoptosis was induced in neuronal cells in same area as iron localization. Significant increases were observed in 4-HNE, HO-1, IL-6, IL-1β, and TNFα mRNA in the hippocampus and hypothalamus. In control, those changes were not observed. Given together, inflammation and oxidative changes were induced by icy injection of Fe-NTA. And since the hippocampus was damaged remarkably, it may be vulnerable to oxidative stress caused by Fe-NTA.