HET0016通过抑制创伤性颅脑损伤后的过度自噬减轻脑损伤

HET0016 alleviates brain injury by inhibiting excessive autophagy after traumatic brain injury

目的探讨20羟基二十碳四烯酸(20-HETE)的清除剂HET0016在创伤性脑损伤(TBI)后对自噬的作用,并解释其中潜在的机制。方法采用可控皮质冲击法对小鼠进行TBI造模,将36只小鼠随机分为Sham组、TBI组和TBI+HET0016组。采用ELISA测定损伤皮质周边的20-HETE表达水平;神经功能学评分评估各组小鼠的神经功能障碍情况;通过测定脑含水量评估各组小鼠脑组织水肿情况;用TUNEL法检测损伤周边的凋亡分子;用免疫荧光法研究损伤周边微管相关蛋白轻链3的表达。结果HET0016能明显减少TBI引起的20-HETE表达上升(P<0.01),还可以缓解神经功能障碍和脑水肿(P<0.01或P<0.05);HET0016处理也减轻了TBI损伤周边组织的细胞凋亡以及过度自噬(P<0.01)。结论HET0016可以抑制过度自噬和细胞凋亡,并改善TBI后的神经功能障碍。

Objective To explore the effect of 20-hydroxyeicosatetraenoic acid(20-HETE)scavenger HET0016 on autophagy after traumatic brain injury(TBI)and explain the underlying mechanism.Methods The controlled cortical impact method was used to establish mouse models with TBI.Thirty-six mice were randomly divided into Sham group,TBI group and TBI+HET0016 group.ELISA was used to measure the expression level of 20-HETE around the injured cortex.Neurological scores were applied to assess the neurological dysfunction of mice in each group.Brain water content was determined to assess the brain tissue edema of mice in each group.TUNEL was performed to detect the apoptotic molecules around the injury.The expression of microtubule-associated protein 1 light chain 3 around the injury was studied by immunofluorescence.Results HET0016 significantly reduced the increased expression of 20-HETE induced by TBI(P<0.01).In addition,HET0016 alleviated neurological dysfunction and cerebral edema(P<0.01 or P<0.05).TUNEL staining showed that HET0016 reduced cell apoptosis in TBI-surrounding tissues and inhibited excessive autophagy(P<0.01).Conclusion HET0016 can inhibit excessive autophagy and apoptosis to improve neurological dysfunction after TBI.