Effect of mild hypothermia on nerve regeneration microenvironment of infarcted area in rat models of cerebral infarction

doi:10.3969/j.issn.2095-4344.2016.27.009

Abstract

Abstract:

BACKGROUND: Numerous studies have demonstrated that mild hypothermia has a better protective effect on neurons after cerebral infarction.
OBJECTIVE: To investigate the effect of mild hypothermia on nerve regeneration microenvironment of infarcted area in rat models of cerebral infarction and analyze its possible effects on neural functional recovery after cerebral infarction.
METHODS: Twenty out of 65 adult female Sprague-Dawley rats were randomly selected as the sham group. The remaining 45 rats were subjected to carotid artery ligation to establish rat models of cerebral infarction. Five rats were rejected because of modeling failure or death, the remaining 40 rats were randomly and evenly divided into cerebral infarction and mild hypothermia groups. The head temperature of rats in the cerebral infarction group was downregulated to (37±1) ℃ using a semiconductor refrigeration instrument. The rats were transferred to the room with the temperature of 25 ℃ after the operation. Brain hypothermia was also induced in rats from the mild hypothermia group. At 13.0-14.0 minutes after establishing rat models of cerebral ischemia, the head on the side of cerebral ischemia was tightly connected with the probe of the semiconductor refrigeration instrument. The refrigerator temperature was adjusted to 6-8 ℃, so as to make the temperature of brain tissue on the lesion side at 32.0-33.0 ℃ for 4 hours.
RESULTS AND CONCLUSION: Compared with the cerebral infarction group, the BBB scores of rats in the mild hypothermia group were distinctly increased, and the volume of infarcted area decreased. At 1 day after modeling, the expression level of growth associated protein 43 mRNA in brain tissue of rats in the mild hypothermia group was close to that in the cerebral infarction group. At 2 weeks after modeling, the expression level of growth associated protein 43 mRNA in brain tissue of rats in the mild hypothermia group was significantly increased compared with that in the cerebral infarction group. These results suggest that mild hypothermia therapy can protect nerve cells against injury caused by cerebral infarction and promote the recovery of neurological function. Its underlying mechanism may be related to the up-regulation of the expression of growth associated protein 43 in ischemic penumbra.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Infarction, Middle Cerebral Artery, Hypothemia, GAP-43 Protein, Apoptosis

CLC Number:

R318

Zheng Rui-juan, Gao Yu-hong. Effect of mild hypothermia on nerve regeneration microenvironment of infarcted area in rat models of cerebral infarction[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(27): 4013-4019.

Figures/Tables 3

2.1 造模成功动物数量及过程从65只大鼠中随机取20只作为假手术组，其余45只结扎颈动脉建立脑梗死模型，排除造模失败及死亡的5只，余下40只大鼠随机等分为脑梗死组及亚低温组。最终共60只大鼠进入结果分析。造模流程图见图1。 2.2 模型创新性结扎颈内动脉制备的局灶性脑缺血大鼠模型可以选择性地阻断进入大脑中动脉的血流，阻断颈内动脉的下丘脑分支，仅少量出现下丘脑梗死，更符合临床情况。 2.3 模型更接近临床实践实验以结扎颈动脉方法建立脑梗死模型，经TTC染色确认脑梗死灶明显。 2.4 脑组织病理变化苏木精-伊红染色显示，假手术组大鼠脑组织未见明显病理改变；脑梗死组大鼠脑组织可见大面积梗死灶，神经元出现典型的缺血性改变，在梗死区周围及梗死区可见散在胶质细胞增生，病变部位与周围组织界限较清楚，其中心区神经元数量明显减少；亚低温组大鼠脑组织较脑梗死组梗死灶明显减小，梗死灶内残存的神经元较多，可见轻度缺血性改变(图2)。 2.5 运动功能造模前各组大鼠BBB评分差异无显著性意义(P > 0.05)。与假手术组相比，脑梗死组大鼠BBB评分降低(P < 0.05)。治疗2周后，亚低温组大鼠BBB评分较脑梗死组显著增加(P < 0.05；表2)。 2.6 脑梗死体积 TTC染色发现，假手术组大鼠脑组织未见梗死灶，脑梗死组大鼠梗死灶主要位于皮质和纹状体，而亚低温组大鼠梗死灶仅限于纹状体，且亚低温组大鼠梗死灶体积小于脑梗死组(P < 0.05；表3)。 2.7 脑组织中生长相关蛋白43 mRNA表达反转录PCR检测结果显示，造模后1 d，亚低温组大鼠脑组织生长相关蛋白43 mRNA表达水平与脑梗死组接近(P > 0.05)，2周时亚低温组大鼠脑组织生长相关蛋白43 mRNA表达较脑梗死组显著升高(P < 0.05；图3，表4)。"

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