虾青素对急性脑出血小鼠的神经功能的保护及机制研究

Neuroprotective effect of astaxanthin on acute cerebral hemorrhage in mice and possible mechanism

目的观察虾青素(Astaxanthin;AST)对脑出血(ICH)小鼠脑组织炎症损伤后神经保护作用及机制研究。方法体外原代神经元细胞培养实验观察虾青素对血红蛋白(hemoglobin,Hb)诱导的原代神经元细胞损伤的影响;体内实验小鼠随机分为4组:假手术组、ICH模型组、虾青素治疗组(10 mg/kg和30 mg/kg),在ICH模型组和虾青素治疗组均采用小鼠脑立体定位仪尾状核注射自体血以建立小鼠ICH模型,假手术组注射等量生理盐水。虾青素治疗组于2 h、12 h、24 h、48 h间断灌胃给药,另2组以相同时间灌胃给予等量橄榄油处理,术后1 d、3 d、5 d、7 d对各组小鼠进行神经功能缺损评分(neurological deficit score,NDS),分别在给药3 d和7 d后取脑组织,应用Neu N染色观察小鼠脑组织神经元数量的病理变化,ELISA法检测脑血肿周围组织中的炎症细胞因子,包括白细胞介素IL-1β、凋亡相关因子Caspase-3和肿瘤坏死因子TNF-α的蛋白含量的在ICH中时间趋势的变化。结果 1 d、3 d,10 mg/kg和30 mg/kg虾青素都能减少ICH小鼠的NDS,虾青素治疗组中的ICH小鼠血肿周围组织中的IL-1β、Caspase-3和TNF-α蛋白表达明显低于ICH模型组(P<0.05),虾青素改善了ICH血肿造成的神经元细胞的继发性炎性损伤,进而抑制了神经元细胞凋亡。结论虾青素对ICH小鼠继发炎性损伤具有较好的神经元细胞保护的作用,其潜在的作用机制可能与虾青素改善ICH小鼠神经功能障碍,通过抑制炎症和凋亡的信号传导通路的过度激活,减少了炎症和凋亡相关蛋白持续的表达有关。

Objective To investigate the neuroprotective effect and mechanism of AST on acute cerebral hemorrhage in mice. Methods To investigate the effect of AST on the injury of primary cortex neurons induced by hemoglobin in vitro;in vivo,mices were divided into four groups randomly：sham group,ICH model group,AST group （10 and 30 mg/kg）. The operations of model group and AST groups were guarded by stereotaxic instrument ascertaining the location and injected collagenase VII to induce ICH model,the sham operation was injected the same amount of saline. AST were consecutively gavaged with AST group at 2h,12h,24h and 48h,the other groups were dosed with the same amount of saline. Neurological deficit were scored at 1d,3d,5d and 7d after operation,the total brain tissues were collected at 3d and 7d after administration respectively,pathological changes of brain tissue were observed by NeuN staining in mice,ELISA essay was used to detect the expressions of inflammatory cytocine in perihematomal brain tissue of ICH mice,such as IL-6,Caspase-3 and TNF-α. Results AST （10 mg/kg and 30 mg/kg） reduced the NDS in ICH mice at 1 d and 3 d,the expression of IL-6,Caspase-3 and TNF-α in perihematomal brain tissue of Ica group were less than sham group （P〈0.05 ）. AST attenuated the secondary injury of hematoma,and inhibited the apoptosis of neuron. Conclusion AST provides apparently protective effect against ICH secondary injury in mice. The mechanism may be related to the neuron protection against ICH inflammatory injury,alleviating the neurological deficit of mice,and inhibiting the expression of neuroinflammatory cytocines and apoptosis protein.