实验性蛛网膜下腔出血后早期铁调素表达的变化

Expression of Hepcidin in rats after subarachnoid hemorrhage.

目的观察大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期海马铁含量及铁调素(Hepcidin)蛋白的表达变化,探讨SAH后早期Hepcidin参与脑铁代谢紊乱的可能机制。方法 90只成年雄性SD大鼠,随机分为假手术对照组和蛛网膜下腔出血组。采用视交叉前池单次注血法构建大鼠SAH模型。在SAH后早期12、24、48、72h采用免疫组化、Western blot法检测大鼠SAH后海马组织内Hepcidin的蛋白表达情况,采用原子吸收光谱仪测定海马组织总铁含量。结果免疫组化实验发现,与假手术对照组(0.19±0.05)比较,大鼠SAH后12h(0.30±0.06)见Hepcidin表达水平明显增高,至72h(0.56±0.07)呈逐渐增高趋势,差异有统计学意义(F=31.911,P‐0.05);Western blot实验同样发现,与假手术对照组(0.238±0.047)比较,大鼠SAH后12h(0.481±0.065)见Hepcidin蛋白表达水平明显增高,至72h(1.627±0.143)呈逐渐增高趋势,差异有统计学意义(F=147.314,P‐0.05);原子吸收光谱仪检测大鼠蛛网膜下腔出血后12h海马总铁含量(58.50±9.19)开始增加,至72h(99.34±7.68)呈逐渐增高趋势,与假手术对照组(43.51±4.59)比较,差异有统计学意义(F=28.799,P‐0.05);大鼠海马组织Hepcidin蛋白的表达量与海马铁含量成正相关(r=0.914,P<0.05)。结论大鼠SAH后脑铁含量的增加可能与Hepcidin表达增高有关。

Objective To examine the iron content and the expression of hepcidin in early period after subarach-noid hemorrhage （SAH） in rats, and to explore the role of hepcidin in dysregulation of brain iron metabolism after SAH. Methods Totally 90 adult male SD rats were randomly divided into two groups：the sham-operation group and SAH group. The SAH model was established by single blood injection to prechiasmatic cistern. Immunohistochemical and Western Blotting were used to examine the expression of hepcidin at 12, 24, 48 and 72h after SAH. Meanwhile, Atomic Absorption Spectrometer was used to detect the iron content. Results Immunochemistry showed that hepcidin expression in rats in SAH group began to rise at 12 h（0.30±0.06）and gradually increased over time until 72 h（0.56±0.07）compared with the sham group（0.19±0.05）. The expression of hepcidin was significantly higher in SAH group than in the sham group（F=31.911, P〈0.05）. Western blot showed that hepcidin expression in rats in SAH group began to rise at 12h（0.481±0.065） and gradually increased over time until 72h（1.627±0.143）Compared with the sham group（0.238±0.047）. The expression＆amp;nbsp;of hepcidin was significantly higher in SAH group than in the sham group after SAH（F=147.314,P〈0.05）. Iron content in SAH group began to rise at 12h after SAH（58.50±9.19）and gradually increased until 72 h（99.34±7.68）. The iron con-tents in SAH group were higher at every time points than those in sham group（43.51±4.59）（F=28.799,P﹤0.05）. The ex-pression of hepcidin was correlated with the iron content in SAH group（r=0.914,P﹤0.01）. Conclusion The increase in iron content following SAH is associated with the increased hepcidin expression.