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

去年，将Fe-NTA注射到腹膜液中。在这一年中，铁NTA与脑室内（icv）注射给药，以避免对周围组织的影响。用渗透泵和脑灌注盒向侧脑室持续注入Fe-NTA溶液（Fe0.1mg/ml，0.25 μl/hr），观察脑组织的变化。在对照组中，以相同的方式通过冰冷注射施用生理盐水溶液。给药2周后，取下丘脑、大脑皮质、海马和纹状体，采用实时荧光定量PCR检测血红素氧合酶（HO）-1、IL-6和超氧化物歧化酶（SOD）2 mRNA的表达。HO-1和IL-6 mRNA水平升高，SOD-2 mRNA水平稳定. Fe-NTA给药4周后，脑切片在HE、Pearl染色和TUNEL中染色。定量检测海马和下丘脑中4-羟基-2-壬烯醛（4-HNE）含量，实时荧光定量PCR检测海马和下丘脑中HO-1、IL-6、TNFα、SOD 1、SOD 2、SOD 3、Bc 12、Bad和FasL的mRNA。HE染色可见海马明显受损，并被炎性组织所取代。铁位于炎症组织和正常组织之间。在与铁定位相同的区域中诱导神经元细胞凋亡。海马和下丘脑中4-HNE、HO-1、IL-6、IL-1β和TNFα mRNA表达显著增加。在对照组中，未观察到这些变化。同时给予，通过冰冷注射Fe-NTA诱导炎症和氧化变化。由于海马损伤明显，可能易受Fe-NTA引起的氧化应激的影响。

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.