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目的探讨去铁敏预处理对星型胶质细胞(AS)缺氧损伤的保护作用及可能机制。方法体外培养AS,建立去铁敏糖氧剥夺(OGD)模型,细胞分为:正常培养组、去铁敏预处理组(先给予去铁敏预处理,再给予去铁敏OGD)、OGD组(给予去铁敏OGD)。采用细胞活力测定、细胞核固缩比率、形态学改变评价去铁敏预处理后的保护效应。用免疫荧光染色检测去铁敏预处理后AS的缺氧诱导因子-1α(HIF-1α)和促红细胞生成素(EPO)蛋白表达情况,RT-PCR检测HIF-1α和EPO的mRNA变化情况。结果去铁敏预处理组细胞形态保持良好,AS活力下降减轻为58%(OGD组25%,P<0.05),细胞核固缩百分比为38%(OGD组30%,P<0.05)。免疫荧光染色发现,体外培养的AS在预处理后出现HIF-1α和EPO蛋白表达。RT-PCR发现去铁敏化学缺氧能上调HIF-1α及EPO mRNA表达。结论去铁敏预处理有确切有效的抗缺氧损伤作用,这种效应与保护AS有关,其机制可能是去铁敏诱导了HIF-1α和EPO表达增加。
Objective To investigate the protective effect of desferoxamine preconditioning on hypoxic injury induced by astrocytes (AS) and its possible mechanism. Methods AS was established in vitro and the model of deoxyhemoglobin (OGD) was established. The cells were divided into four groups: normal group, deferoxamine pretreatment group (pretreated with deferoxamine and then deferoxamine OGD), OGD group (Given deferoxib OGD). The protective effect of dextrin pretreatment was evaluated by cell viability assay, nuclear condensation ratio and morphological changes. The expression of hypoxia inducible factor-1α (HIF-1α) and erythropoietin (EPO) protein in AS after deferoxamine preconditioning was detected by immunofluorescence staining. The changes of mRNA of HIF-1α and EPO were detected by RT-PCR. Results The cell morphology of deferoxamine pretreatment group was well maintained. The reduction of AS activity was 58% (OGD group 25%, P <0.05). The percentage of nuclear pyknosis was 38% (OGD group 30%, P <0.05). Immunofluorescence staining showed that AS cultured in vitro showed HIF-1α and EPO protein expression after pretreatment. RT-PCR showed that deoxynil hypoxia upregulated the expression of HIF-1α and EPO mRNA. Conclusion The dextrin pretreatment has an exact and effective anti-hypoxic injury, which is related to the protection of AS. The possible mechanism is that deferoxamine induces the increase of HIF-1α and EPO expression.