高压氧修复缺血再灌注损伤大鼠血脑屏障的自噬机制

doi:10.3969/j.issn.1006-9771.2022.04.007

Abstract

Abstract:

Objective To observe the effect of hyperbaric oxygen chamber treatment on the expression of autophagy protein microtubule associated protein 1 light chain 3 (LC3) and Beclin-1 in cerebral vascular endothelial cells of rats with ischemia-reperfusion injury, and to explore the mechanism of hyperbaric oxygen in repairing blood-brain barrier in rats with ischemia-reperfusion injury.Methods A total of 54 Sprague-Dawley rats were randomly divided into sham operated group (n=12), cerebral ischemia-reperfusion injury (CIRI) model group (n=18), hyperbaric oxygen group (n=12) and inhibitor group (n=12). The CIRI model was made by a suture method in the model group, hyperbaric oxygen group and inhibitor group. The hyperbaric oxygen group and inhibitor group accepted hyperbaric oxygen, and the inhibitor group injected 3-methyladenine in lateral cerebral ventricle before treatment. They were stained with Evans blue and Evans blue content in the infarct area was detected 72 hours after CIRI. Double immunofluorescent staining was used to observe the expression of LC3 in vascular endothelial cells marked with CD₃₁ in the model group, and LC3-II and Beclin-1 in infarcted cortical microvascular were examined by Western blotting in all the groups.Results Compared with the sham operation group, Evans blue content in the infarct area was significantly higher in the model group (P<0.01); compared with the model group, the content of Evans blue in the infarct area decreased significantly in hyperbaric oxygen group (P<0.01); compared with the hyperbaric oxygen group, the content of Evans blue in the infarct area was significantly higher in the inhibitor group (P < 0.05). There was obvious expression of LC3 in CD₃₁⁺ cells. The expression levels of Beclin-1 and LC3-Ⅱ in the infarcted area were significantly higher in the model groupt than in the sham operation group (P<0.01); and it was significantly higher in the hyperbaric oxygen group than in the model group (P<0.05) and was lower in the inhibitor group than in the hyperbaric oxygen group and model group (P < 0.01).Conclusion Autophagy exists in vascular endothelial cells in the injured area of rats with CIRI. Hyperbaric oxygen can upregulate the expression of autophagy proteins LC3-Ⅱ and Beclin-1 in vascular endothelial cells in the infarcted area, to promote repairing blood-brain barrier.

Key words: hyperbaric oxygen, ischemia-reperfusion injury, autophagy, brain endothelial cell, blood-brain barrier

HUANG Feifei,WANG Wansong,DONG Xiaoyang,FENG Zhen. Repair of blood-brain barrier by hyperbaric oxygen via autophagy in rats with cerebral ischemia-reperfusion injury[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(4): 415-420.

Figures/Tables 4

References 35

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组别	n	EB含量
假手术照组	6	0.887±0.191
模型组	6	2.423±0.223^a
高压氧组	6	1.989±0.163^b
抑制剂组	6	2.249±0.174^c
F值		79.726
P值		< 0.001

组别	n	LC3-Ⅱ/LC3-Ⅰ	Beclin-1/β-actin
假手术组	6	0.255±0.041	0.873±0.174
模型组	6	0.385±0.064^a	1.235±0.090^a
高压氧组	6	0.487±0.066^b	1.549±0.660^b
抑制剂组	6	0.268±0.082^c,d	0.934±0.048^c,d
F值		16.927	51.700
P值		< 0.001	< 0.001

Repair of blood-brain barrier by hyperbaric oxygen via autophagy in rats with cerebral ischemia-reperfusion injury

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