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目的探究Neuritin对大鼠蛛网膜下腔出血(SAH)后早期脑损伤过程中内质网应激因子GRP78及血-脑脊液屏障通透性的影响。方法 2014年4月—2015年9月选取成年健康雄性SD大鼠144只,按照随机数字表法分为对照组、假手术组(Sham组)、SAH组、Neuritin组,各36只。再依据简单随机抽样法分别于3、6、12、24、48、72 h抽取每组6只大鼠处死,Neuritin组于处死前1 h采用立体定向技术向侧脑室注射Neuritin。建模后6、12、24、48、72 h处死大鼠前,记录各组大鼠Garcia神经功能评分;建模后3、6、12、24、48、72 h处死大鼠后,采用HE染色观察SAH建立情况,侧脑室微量注射亚甲蓝鉴定Neuritin注射情况;采用Western blotting法检测大鼠大脑皮质内质网应激因子GRP78表达水平;采用甲酰胺浸泡法检测大鼠脑内伊文蓝(EB)含量以评价血-脑脊液屏障通透性。结果 SAH组、Neuritin组建模后6、12、24、48、72 h大鼠Garcia神经功能评分均低于对照组和Sham组(P<0.05);Neuritin组建模后6、12、24、48、72 h大鼠Garcia神经功能评分均高于SAH组(P<0.05)。SAH组大鼠大脑表面弥散分布积血,其中在willis环、小脑延髓池及脑干腹侧有大量积血存在。Neuritin组大脑腹面有亚甲蓝循环存在,大脑背面无亚甲蓝,Neuritin侧脑室注射成功。SAH组建模后3、6、12、24、48、72 h GRP78表达水平高于对照组、Sham组(P<0.05);Neuritin组建模后6、12、24、48、72 h GRP78表达水平低于SAH组(P<0.05)。SAH组大鼠建模后3、6、12、24、48、72 h脑组织EB含量均高于对照组、Sham组(P<0.05);Neuritin组大鼠建模后3、12 h脑组织EB含量高于SAH组,建模后6、48、72 h脑组织EB含量低于SAH组(P<0.05)。结论 SAH后早期脑损伤过程中存在内质网应激反应,神经营养因子Neuritin可显著改变内质网应激因子GRP78表达水平,改善早期脑损伤过程中大鼠的神经行为学评分,并对血-脑脊液屏障通透性产生影响。
Objective To investigate the effect of Neuritin on endoplasmic reticulum stress factor GRP78 and blood-cerebrospinal fluid barrier permeability in early brain injury following subarachnoid hemorrhage (SAH) in rats. METHODS: From April 2014 to September 2015, 144 healthy male SD rats were selected and divided into control group, sham operation group (Sham group), SAH group and Neuritin group according to random number table. Six rats in each group were sacrificed at 3, 6, 12, 24, 48, and 72 h according to a simple random sampling method. The Neuritin group was injected with Neuritin into the lateral ventricle by stereotactic technique 1 h before sacrifice. The rats were sacrificed at 6, 12, 24, 48 and 72 h after modeling, and the neurological function scores of Garcia were recorded. After the rats were killed at 3, 6, 12, 24, 48 and 72 h after modeling, The establishment of SAH was observed by staining, and the injection of Neuritin in the lateral ventricle was detected by microinjection of methylene blue. The expression of GRP78 in the cerebral cortex was detected by Western blotting. The expression of GRP78, EB) content to assess blood-brain barrier permeability. Results The neurological scores of Garcia in SAH group and Neuritin group were lower than those in control group and Sham group at 6, 12, 24, 48 and 72 h after modeling (P <0.05) At 48 and 72 h, the neurological function of Garcia was higher than that of SAH group (P <0.05). SAH group diffusely distributed hemorrhage on the surface of the brain, including a large amount of hemorrhage in the willis ring, cerebellar cistern and ventral brainstem. Neuritin group had ventral circulation of methylene blue, no methylene blue on the back of brain, Neuritin lateral ventricle injection was successful. GRP78 expression in SAH group was higher than that in control group and Sham group at 3, 6, 12, 24, 48 and 72 h after model establishment (P <0.05); GRP78 expression in Neuritin group at 6, 12, The level was lower than SAH group (P <0.05). The content of EB in brain of SAH group was higher than that of control group and Sham group at 3, 6, 12, 24, 48 and 72 h after modeling (P <0.05) EB content was higher in SAH group than in SAH group (P <0.05) at 6, 48 and 72 h after model establishment. Conclusions Endoplasmic reticulum stress response exists in the process of early brain injury after SAH. Neuritin, a neurotrophic factor, can significantly change the expression of endoplasmic reticulum stress factor GRP78 and improve neurobehavioral scores of rats during early brain injury. - Cerebrospinal fluid barrier permeability effects.