热休克蛋白A5介导的自噬在小鼠脑缺血/再灌注损伤中的作用

doi:10.12047/j.cjap.5472.2017.058

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (3): 234-238.doi: 10.12047/j.cjap.5472.2017.058

热休克蛋白A5介导的自噬在小鼠脑缺血/再灌注损伤中的作用

彭志锋¹, 孟健², 张继红¹

1. 山西大同大学医学院生理学教研室, 大同 037009;
2. 山西大同大学医学院人体解剖学教研室, 大同 037009

收稿日期:2016-07-12 修回日期:2017-01-23 出版日期:2017-05-28 发布日期:2018-06-20
通讯作者: 彭志锋,Tel:15910699956,E-mail:pzf181@126.com E-mail:pzf181@126.com
基金资助:
山西省基础研究项目（2015021178）；山西大同大学博士科研启动经费（2014-B-01）

Effects of heat shock protein A5 induced autophagy on cerebral ischemia/reperfusion injury in mice

PENG Zhi-feng¹, MENG Jian², ZHANG Ji-hong¹

1. Department of Physiology, School of Medicine, Shanxi Datong University, Datong 037009, China;
2. Department of Anatomy, School of Medicine, Shanxi Datong University, Datong 037009, China

Received:2016-07-12 Revised:2017-01-23 Online:2017-05-28 Published:2018-06-20
Supported by:
山西省基础研究项目（2015021178）；山西大同大学博士科研启动经费（2014-B-01）

摘要/Abstract

摘要： 目的：探讨热休克蛋白A5（HSPA5）诱导的自噬在小鼠脑缺血/再灌注损伤中的作用。方法：将36只BALB/c小鼠随机分为sham、缺血再灌注（I/R）、vehicle + I/R、3-甲基腺嘌呤（3-MA） + I/R、scramble siRNA + I/R和HSPA5 siRNA + I/R组（n=6）。Sham组只进行手术操作，不插入线栓。I/R采用大脑中动脉阻塞（MCAO）60 min后再灌注24 h。Vehicle + I/R组和3-MA + I/R将5μl 0.9% NaCl或3-MA （30 mg/ml）在MCAO前30 min侧脑室注射。scramble siRNA + I/R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。检测神经细胞内自噬体、缺血大脑皮层（LC3）-Ⅱ/LC3-I表达、神经元损伤程度及神经功能缺损。结果：显微镜下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，自噬体形成。与sham组比较，I/R组缺血大脑皮层LC3-Ⅱ/LC3-I蛋白表达水平显著增高（P < 0.05）；与I/R组相比，3-MA + I/R组或HSPA5 siRNA + I/R组缺血大脑皮层LC3-Ⅱ/LC3-I蛋白表达明显减少（P < 0.05）；3-MA + I/R组及HSPA5 siR-NA + I/R组I/R后脑缺血性损伤及神经系统症状加重（P < 0.05）。结论：HSPA5诱导自噬可能在小鼠局灶性I/R损伤中发挥保护作用。

关键词: 热休克蛋白A5, 自噬, 缺血/再灌注损伤, 小鼠

Abstract: Objective: To determine the role of heat shock protein A5 (HSPA5) induced autophagy on cerebral ischemia/reperfusion in-jury in mice. Methods: Thirty-six BALB/c mice were randomly divided into sham group, ischcmia/reperfusion (I/R) group, vehicle + I/R group, 3-Methyladenine(3-MA) + I/R group, scramble siRNA group and HSPA5 siRNA + I/R group(n=6). In sham group, the operation was only performed, did not insert line switch. Focal cerebral ischemia was performed using the method of middle cerebral artery occlusion (MCAO) for 60 min and 24 h reperfusion. In vehicle + I/R group and 3-MA + I/R group, 2μl 0.9% NaCl or 3-MA(30 mg/ml) was admin-istered by intracerebroventricular injection 30 min before MCAO; In scramble siRNA + I/R group and HSPA5 siRNA + I/R group, 5μl scram-ble siRNA or HSPA5 siRNA(2μg/μl) was administered by intracerebroventricular injection 24 h before MCAO. Autophagosome in neuron, the expression of microtubule-associated protein light chain 3 (LC3)-Ⅱ/LC3-I in ischemic cortex, the degree of cerebral ischemic injury and neu-rological function score were detected. Results: Initial electron microscopy showed that neuronal morphology appeared to be normal in the sham group. At 24 h after I/R, cell shrinkage, loss of cellular organelles and formation of autophagosomes were observed in the ischemic cerebral cortex of I/R group. In addition, autophagosomes were less frequently observed than that in I/R group. The expressions of LC3-Ⅱ/LC3-I and Beclin-1 protein were increased significantly in I/R group compared with that in sham group(P < 0.05). Compare with I/R group, the LC3-Ⅱ/LC3-I protein levels induced by I/R in 3-MA + I/R group or HSPA5 siRNA + I/R group was decreased effectively (P < 0.05). In addi-tion, the cerebral ischemic injury and neurological symptoms after I/R in 3-MA + I/R group or HSPA5 siRNA + I/R group were exacerbated significantly (P < 0.05). Conclusion: These results suggest that HSPA5 induced autophagy may play a protective role in focal I/R damage in mice.

Key words: HSPA5, autophagy, ischemia/reperfusion injury, mice

彭志锋, 孟健, 张继红. 热休克蛋白A5介导的自噬在小鼠脑缺血/再灌注损伤中的作用[J]. 中国应用生理学杂志, 2017, 33(3): 234-238.

PENG Zhi-feng, MENG Jian, ZHANG Ji-hong. Effects of heat shock protein A5 induced autophagy on cerebral ischemia/reperfusion injury in mice[J]. CJAP, 2017, 33(3): 234-238.

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热休克蛋白A5介导的自噬在小鼠脑缺血/再灌注损伤中的作用

Effects of heat shock protein A5 induced autophagy on cerebral ischemia/reperfusion injury in mice

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