比较热休克蛋白A5和3-甲基腺嘌呤和介导小鼠脑缺血再灌注损伤的保护作用

发布时间：2018-01-20 04:15

本文关键词： 热休克蛋白A(HSPA) -甲基腺嘌呤(-MA) 自噬缺血再灌注损伤小鼠　出处：《神经解剖学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的:比较研究热休克蛋白A5(HSPA5)和3-甲基腺嘌呤(3-MA)介导小鼠脑缺血再灌注(I/R)损伤性自噬的保护作用。方法:成年雄性BALB/c小鼠30只,分为6组:sham组、缺血再灌注组(I/R组)、vehicle+I/R组、3-甲基腺嘌呤(3-MA)+I/R组、scramble siRNA+I/R组和HSPA5 siRNA+I/R组,每组5只。采用大脑中动脉阻塞(MCAO)60 min后再灌注24 h制作小鼠I/R模型。Vehicle+I/R组和3-MA+I/R将5μl 0.9%Na Cl或3-MA(30 mg/ml)在MCAO前30 min侧脑室注射;scramble siRNA+I/R group组和HSPA5 siRNA+I/R组将5μl scramble siRNA或HSPA5 siRNA(2μg/μl)在MCAO前24 h侧脑室注射。通过小鼠I/R后神经功能评分确定运动功能障碍程度。小鼠神经细胞内自噬体形成用透射电子显微镜测定,再灌注24 h缺血大脑皮层(LC3)-II/LC3-I表达用Western Blot方法检测。小鼠I/R后缺血大脑神经细胞损伤用Nissl染色进行观察。结果:HSPA5和3-MA可以影响I/R小鼠行为学改变并使脑缺血性损伤和神经症状加重(P0.05)。透射电子显微镜检测可见,sham组小鼠大脑皮层神经细胞形态正常,I/R组小鼠缺血大脑皮层神经元胞质中细胞器减少,形成自噬体。与sham组小鼠比较,I/R组小鼠缺血大脑皮层LC3-II蛋白表达水平显著增高(P0.05);3-MA+I/R或HSPA5 siRNA+I/R小鼠缺血大脑皮层LC3-II蛋白表达水平明显低于I/R小鼠(P0.05)。结论:HSPA5和3-MA在介导小鼠脑缺血再灌注损伤导致的自噬可发挥相同的保护作用,并且促进神经细胞凋亡。
[Abstract]:Objective: to compare the effects of heat shock protein A5 (HSPA5) and 3-methyladenine 3-MA) on cerebral ischemia-reperfusion (I / R) in mice. Methods: 30 adult male BALB/c mice. They were divided into 6 groups: sham group, ischemia reperfusion group, I / R group, I / R group, I / R group) I / R group. Scramble siRNA I / R group and HSPA5 siRNA I / R group. Middle cerebral artery occlusion (MCAO). Mice I / R model. Vehicle I / R group and 3-MA I / R group were perfused with 5 渭 l NaCl or 3-MAR for 24 h after reperfusion for 60 min. 30 mg / ml was injected into the lateral ventricle 30 min before MCAO. The scramble siRNA I / R group group and the HSPA5 siRNA I / R group will be 5 渭 l scramble siRNA or HSPA5. SiRNAs. 2 渭 g / 渭 l). The degree of motor dysfunction was determined by the neurological function score of mice after I / R 24 hours before MCAO. The formation of autophagy in mouse nerve cells was measured by transmission electron microscope. Cerebral cortex (LC3) after 24 h reperfusion. The expression of -II / LC3-I was detected by Western Blot method. The injury of cerebral nerve cells after I / R in mice was observed by Nissl staining. HSPA5 and 3-MA could affect behavioral changes and aggravate ischemic brain injury and neurological symptoms in I / R mice (P < 0.05). P0.05. Transmission electron microscopy can be seen. The morphology of cerebral cortex neurons in sham group was normal. The organelles in ischemic cortex neurons decreased and autophagy was formed in II-R group, compared with sham group. In I / R group, the expression of LC3-II protein in ischemic cerebral cortex was significantly higher than that in control group (P 0.05). The expression of LC3-II protein in ischemic cerebral cortex of 3-MA I / R or HSPA5 siRNA I / R mice was significantly lower than that of I / R mice (P 0.05). Conclusion: HSPA5 and 3-MA may play the same protective effect on autophagy induced by cerebral ischemia-reperfusion injury in mice. And promote neuronal apoptosis.
【作者单位】：临夏州积石山县医院大外科;解放军169医院消化血液科;兰州大学附属第二医院;解放军兰州总医院干部病房;
【基金】：国家自然科学基金(81200247) 甘肃省自然科学基金(120RJZA274)
【分类号】：R743
【正文快照】： 韩世强1△,闵恒2△,任海军3,张新宇4,付学锋4*(1.临夏州积石山县医院大外科,临夏731700;2.解放军169医院消化血液科,衡阳421000;3.兰州大学附属第二医院,兰州730030;4.解放军兰州总医院干部病房,兰州730050)脑缺血/再灌注(ischemia/reperfusion,I/R)损伤是脑部缺血一定时间而

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