丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响

doi:10.3969/j.issn.1006-9771.2021.05.005

《中国康复理论与实践》 ›› 2021, Vol. 27 ›› Issue (5): 522-529.doi: 10.3969/j.issn.1006-9771.2021.05.005

丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响

周文美^1,²,陶陶^2,³(),吴霜¹,王廷龙⁴,杨正奕⁴,张莹^1,²

^1.贵州医科大学附属医院康复医学科，贵州贵阳市 550001
^2.贵州省人民医院康复医学科，贵州贵阳市 550002
^3.贵州医科大学附属人民医院康复医学科，贵州贵阳市 550002
^4.贵州省食品药品检验所，贵州贵阳市 550004

收稿日期:2020-06-09 修回日期:2020-11-27 出版日期:2021-05-25 发布日期:2021-05-26
通讯作者: 陶陶 E-mail:835707237@qq.com
作者简介:周文美(1995-)，女，穿青人，贵州毕节市人，硕士研究生，主要研究方向：脑血管疾病治疗与康复|陶陶(1964-)，女，汉族，贵州贵阳市人，博士，教授、主任医师，硕士生导师，主要研究方向：脑血管疾病治疗与康复。
基金资助:
贵州省科技计划项目(黔科合基础[2019]1207号)

Effects of Enriched Environment on Neurological Function and Glucose Metabolism in Ischemic Penumbra in Cerebral Ischemia-reperfusion Injury Rats

Wen-mei ZHOU^1,²,Tao TAO^2,³(),Shuang WU¹,Ting-long WANG⁴,Zheng-yi YANG⁴,Ying ZHANG^1,²

^1.Department of Rehabilitation Medicine, the Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550001, China
^2.Department of Rehabilitation Medicine, Guizhou Provincial People's Hospital, Guiyang, Guizhou 550002, China
^3.Department of Rehabilitation Medicine, the Affiliated People's Hospital of Guizhou Medical University, Guiyang, Guizhou 550002, China
^4.Guizhou Institute for Food and Drug Control, Guiyang, Guizhou 550004, China

Received:2020-06-09 Revised:2020-11-27 Published:2021-05-25 Online:2021-05-26
Contact: Tao TAO E-mail:835707237@qq.com
Supported by:
Guizhou Science and Technology Plan(2019-1207)

摘要/Abstract

摘要： 目的

探讨丰富环境对脑缺血再灌注损伤大鼠神经功能及缺血半暗带区葡萄糖代谢的影响。

方法

成年雄性Sprague-Dawley大鼠72只随机分为假手术组(n = 24)、模型组(n = 24)和丰富环境组(n = 24)。后两组采用改良Longa法制备大鼠脑缺血60 min再灌注模型。丰富环境组造模后立即饲养在丰富环境中。分别于造模前及造模后1 d、7 d、14 d、21 d、28 d采用改良神经损害严重程度评分(mNSS)进行评估。术后1 d和28 d，神经功能评估后各组取12只，HE染色观察脑组织病理结构，逆转录实时定量聚合酶链反应(RT-qPCR)和Western blotting检测缺血半暗带缺氧诱导因子-1α (HIF-1α)、葡萄糖转运蛋白1 (GLUT1)、6-磷酸果糖-2-激酶/果糖-2,6-双磷酸酶3 (PFKFB3) mRNA和蛋白表达；高效液相色谱法测定缺血半暗带ATP、ADP和AMP变化，计算能量负荷(EC)。

结果

术后14 d后，丰富环境组mNSS评分持续低于模型组(P < 0.05)。术后1 d，与假手术组相比，模型组和丰富环境组缺血半暗带细胞出现水肿、核固缩边缘化、成空泡状排列等病理改变，ATP含量及EC下降(P < 0.05)，HIF-1α、GLUT1和PFKFB3 mRNA和蛋白表达增高(P < 0.05)。术后28 d，与模型组相比，丰富环境组病理改善，缺血半暗带区ATP含量及EC增高(P < 0.05)，HIF-1α、GLUT1和PFKFB3 mRNA和蛋白表达增高(P < 0.05)。

结论

丰富环境可促进脑缺血再灌注损伤大鼠神经功能恢复，可能与上调HIF-1α及其下游基因GLUT1和PFKFB3表达，改善葡萄糖代谢有关。

关键词: 脑缺血再灌注, 丰富环境, 葡萄糖代谢, 缺氧诱导因子-1α, 葡萄糖转运蛋白-1, 6-磷酸果糖-2-激酶/果糖-2,6-双磷酸酶3, 大鼠

Abstract: Objective

To explore the effects of enriched environment on neurological function in cerebral ischemia-reperfusion injury rats and the glucose metabolism in ischemic penumbra.

Methods

A total of 72 adult male Sprague-Dawley rats were randomly divided into sham group (n = 24), model group (n = 24) and enriched environment group (n = 24). The latter two groups suffered cerebral ischemia 60 minutes and reperfused with modified Longa's method. The enriched environment group was fed in enriched environment after operation. All the rats were assessed with modified Neurological Severity Score (mNSS) before, and one, seven, 14, 21 and 28 days after operation. One and 28 days after operation, twelve rats from each groups were sacrificed after mNSS assessment, respectively. The histopathology was observed with HE staining. The expressions of hypoxia-inducible factor-1α (HIF-1α), glucose transporter 1 (GLUT1) and 6-phosphofructo-2-kinase/fructose-2-bisphosphatase 3 (PFKFB3) in ischemic penumbra were determined with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. The levels of ATP, ADP and AMP in ischemic penumbra were measured with high performance liquid chromatography (HPLC) and the energy charge (EC) was calculated.

Results

Compared with the model group, the scores of mNSS decreased in the enriched environment group since 14 days after operation (P < 0.05). The cells in the penumbra presented edema, nuclear pyknosis marginalization, vacuolar arrangement and other pathological changes in the model group and the riched environment group one day after operation; while compared with the model group, the levels of ATP and EC decreased and the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased in the enriched environment group (P < 0.05). The pathology improved in the riched environment group compared with that in the model group 28 days after operation; while the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased, as well as the levels of ATP and EC (P < 0.05).

Conclusion

Enriched environment can promote the recovery of neurological function in rats after cerebral ischemia-reperfusion, which may associate with promoting expression of HIF-1α and downstream GLUT1 and PFKFB3, and improving glucose metabolism.

Key words: cerebral ischemia-reperfusion, riched environment, glucose metabolism, hypoxia inducible factor-1α, glucose carrier-1, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3, rats

中图分类号:

R743.3

周文美,陶陶,吴霜,王廷龙,杨正奕,张莹. 丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响[J]. 《中国康复理论与实践》, 2021, 27(5): 522-529.

Wen-mei ZHOU,Tao TAO,Shuang WU,Ting-long WANG,Zheng-yi YANG,Ying ZHANG. Effects of Enriched Environment on Neurological Function and Glucose Metabolism in Ischemic Penumbra in Cerebral Ischemia-reperfusion Injury Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(5): 522-529.

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参考文献 31

1	PANISELLO-ROSELLÓ A, ROSELLÓ-CATAFAU J. Molecular mechanisms and pathophysiology of ischemia-reperfusion injury [J]. Int J Mol Sci, 2018, 19(12): 4093.
2	ASTRUP J, SIESJÖ B K, SYMON L. Thresholds in cerebral ischemia-the ischemic penumbra [J]. Stroke, 1981, 12(6): 723-725.
3	ZHANG Y J, XU D, QI H, et al. Enriched environment promotes post-stroke neurogenesis through NF-κB-mediated secretion of IL-17A from astrocytes [J]. Brain Res, 2018, 1687: 20-31.
4	BALAMURUGAN K. HIF-1 at the crossroads of hypoxia, inflammation, and cancer [J]. In J Cancer, 2016, 138(5): 1058-1066.
5	MASOUD G N, LI W. HIF-1α pathway: role, regulation and intervention for cancer therapy [J]. Acta Pharm Sin B, 2015, 5(5): 378-389.
6	HU L, ZENG Z, XIA Q, et al. Metformin attenuates hepatoma cell proliferation by dcreasing glycolytic flux through the HIF-1α/PFKFB3/PFK1 pathway [J]. Life Sci, 2019, 239: 116966.
7	邓敏,章军建,沈军,等. 丰富环境对慢性脑低灌注大鼠低氧诱导因子血管内皮生长因子的表达及其认知功能的影响[J]. 中华老年医学, 2016, 35(1): 79-84.
	DENG M, ZHANG J J, SHEN J, et al. Effect of environmental enrichment on cognitive function and the expressions of hypoxia inducible factor 1α and vascular endothelial growth factor in rats with chronic cerebral hypoperfusion [J]. Chin J Geriatr, 2016, 35(1): 79-84.
8	WU X, LIU S, HU Z, et al. Enriched housing promotes post-stroke neurogenesis through calpain 1-STAT3/HIF-1α/VEGF signaling [J]. Brain Res Bull, 2018, 139: 133-143.
9	YAN R Y, WANG S J, YAO G T, et al. The protective effect and its mechanism of 3-n-butylphthalide pretreatment on cerebral ischemia reperfusion injury in rats [J]. Eur Rev Med Pharmacol Sci, 2017, 21(22): 5275-5282.
10	ZHU Y, DENG L, TANG H J, et al. Electroacupuncture improves neurobehavioral function and brain injury in rat model of intracerebral hemorrhage [J]. Brain Res Bull, 2017, 131: 123-132.
11	XIE H, YU K, ZHOU N, et al. Enriched environment elicits proangiogenic mechanisms after focal cerebral ischemia [J]. Transl Stroke Res, 2019, 10(2): 150-159.
12	THIRUGNANACHANDRAN T, MA H, SINGHAL S, et al. Refining the ischemic penumbra with topography [J]. Int J Stroke, 2018, 13(3): 277-284.
13	ZHAN C, YANG J, ZHAN L. RP-HPLC determination of effects of isoliquiritigenin on brain energy metabolism in repeated cerebral ischemia-reperfusion mice [J]. Chin J Pharm Anal, 2005, 25: 639-642.
14	PANISELLO-ROSELLÓ A, ROSELLÓ-CATAFAU J. Molecular mechanisms and pathophysiology of ischemia-reperfusion injury [J]. Int J Mol Sci, 2018, 19 (12): 4093.
15	MALÁ H, RASMUSSEN C P. The effect of combined therapies on recovery after acquired brain injury: systematic review of preclinical studies combining enriched environment, exercise, or task-specific training with other therapies [J]. Restor Neurol Neuros, 2017, 35(1): 25-64.
16	吕富岩,张雷红,宫兆帅,等. 丰富环境促进缺氧缺血性脑损伤神经可塑性的研究进展[J]. 中国康复理论与实践, 2018, 24(5): 509-512.
	LÜ F Y, ZHANG L H, GONG Z S, et al. Advance of enriched environment in neural plasticity post hypoxic-ischemic brain damage (review) [J]. Chin J Rehabil Theory Pract, 2018, 24(5): 509-512.
17	李敏,陶陶,张继荣. 丰富环境对脑缺血再灌注损伤后相关凋亡基因的研究进展[J]. 中国康复医学杂志, 2018, 33(10): 1238-1241.
	LI M, TAO T, ZHANG J R. Chin J Rehabil Med, 2018, 33(10): 1238-1241.
18	WANG D, YUAN X, LIU T, et al. Neuroprotective activity of lavender oil on transient focal cerebral ischemia in mice [J]. Molecules, 2012, 17(8): 9803-9817.
19	KANG H, CHOI D H, KIM S K, et al. Alteration of energy metabolism and antioxidative processing in the hippocampus of rats reared in long-term environmental enrichment [J]. Dev Neurosci, 2016, 38(3): 186-194.
20	BRIONES T L, WOODS J, ROGOZINSKA M. Decreased neuroinflammation and increased brain energy homeostasis following environmental enrichment after mild traumatic brain injury is associated with improvement in cognitive function [J]. Acta Neuropathol Com, 2013, 1: 57.
21	UZDENSKY A B. Regulation of apoptosis in the ischemic penumbra in the first day post-stroke [J]. Neural Regen Res, 2020, 15(2): 253-254.
22	AMARAL A I, TEIXEIRA A P, MARTENS S, et al. Metabolic alterations induced by ischemia in primary cultures of astrocytes:merging 13C NMR spectroscopy and metabolic flux analysis [J]. Neurochem, 2010, 113(3): 735-748.
23	MARIN-VALENCIA I, GOOD L B, MA Q, et al. Glut1 deficiency (G1D): epilepsy and metabolic dysfunction in a mouse model of the most common human phenotype [J]. Neurobiol Dis, 2012, 48(1): 92-101.
24	WU X, WANG C, WANG J, et al. Hypoxia preconditioning protects neuronal cells against traumatic brain injury through stimulation of glucose transport mediated by HIF-1α/GLUTs signaling pathway in rat [J]. Neurosurg Rev, 2021, 44(1): 411-422.
25	LV Y, ZHANG B, ZHAI C, et al. PFKFB3-mediated glycolysis is involved in reactive astrocyte proliferation after oxygen-glucose deprivation/reperfusion and is regulated by Cdh1 [J]. Neurochem Int, 2015, 91: 26-33.
26	LI C, ZHANG B, ZHU Y, et al. Post-stroke constraint-induced movement therapy increases functional recovery, angiogenesis, and neurogenesis with enhanced expression of HIF-1α and VEGF [J]. Curr Neurovasc Res, 2017, 14(4): 368-377.
27	ZHANG Z, YAN J, SHI H. Role of hypoxia inducible factor 1 in hyperglycemia-exacerbated blood-brain barrier disruption in ischemic stroke [J]. Neurobiol Dis, 2016, 95: 82-92.
28	SADLECKI P, BODNAR M, GRABIEC M, et al. The role of hypoxia-inducible factor-1α, glucose transporter-1 (GLUT-1) and carbon anhydrase IX in endometrial cancer patients [J]. Biomed Res Int, 2014, 2014: 616850.
29	FUKASAWA M, TSUCHIYA T, TAKAYAMA E, et al. Identification and characterization of the hypoxia-responsive element of the human placental 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase gene [J]. J Biochem, 2004, 136(3): 273-277.
30	VRIEND J, REITER R J. Melatonin and the von Hippel-Lindau/HIF-1 oxygen sensing mechanism: a review [J]. Biochim Biophys Acta, 2016, 1865(2): 176-183.
31	金信浩,郑兵,吴雪莲,等. 丰富环境对脑卒中患者神经功能康复的效果[J]. 中国康复理论与实践, 2017, 23(3): 323-325.
	JIN X H, ZHENG B, WU X L, et al. Effects of environmental enrichment on neurological functions of patients with stroke [J]. Chin J Rehabil Theory Pract, 2017, 23(3): 323-325.

组别	n	0 d	1 d	7 d	14 d	21 d	28 d
假手术组	8	0	0	0	0	0	0
模型组	8	0	10.38±0.92^a	9.25±0.71^a	8.63±0.74^a	7.75±1.04	7.13±0.64^a
丰富环境组	8	0	10.50±0.93^a	9.13±0.99^a	7.88±0.83^a,b	6.88±0.64^a,b	5.63±0.92^a,b
F值			513.800	455.675	438.300	291.723	270.900
P值			< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	n	HIF-1α		GLUT1		PFKFB3
组别	n	1 d	28 d	1 d	28 d	1 d	28 d
假手术组	6	0.02±0.02	0.02±0.03	0.01±0.02	0.01±0.02	0.01±0.01	0.03±0.04
模型组	6	0.79±0.19^a	0.10±0.01^a	0.64±0.04^a	0.14±0.06^a	2.25±0.29^a	0.28±0.03^a
丰富环境组	6	0.81±0.18^a	0.24±0.04^a,b	0.62±0.09^a	0.23±0.04^a,b	2.10±0.24^a	0.87±0.14^a,b
F值		27.281	46.799	119.965	18.401	127.415	73.408
P值		0.001	< 0.001	< 0.001	0.003	< 0.001	< 0.001

组别	n	HIF-1α		GLUT1		PFKFB3
组别	n	1 d	28 d	1 d	28 d	1 d	28 d
假手术组	6	0.38±0.04	0.43±0.03	0.41±0.08	0.48±0.08	0.51±0.05	0.41±0.06
模型组	6	1.22±0.11^a	0.75±0.08^a	1.24±0.08^a	0.75±0.03^a	1.58±0.10^a	1.04±0.12^a
丰富环境组	6	1.27±0.09^a	0.93±0.07^a,b	1.18±0.06^a	0.88±0.06^a,b	1.56±0.05^a	1.31±0.06^a,b
F值		105.196	44.379	121.472	41.537	236.893	91.819
P值		< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	n	ATP(μg/g)		ADP(μg/g)		AMP(μg/g)		EC
组别	n	1 d	28 d	1 d	28 d	1 d	28 d	1 d	28 d
假手术组	3	16.15±1.31	17.40±0.56	215.09±4.31	167.91±1.15	49.32±1.67	49.10±0.60	0.44±0.00	0.43±0.00
模型组	3	4.91±0.88^a	9.95±0.38^a	71.97±2.24^a	67.81±0.03^a	105.93±1.27^a	38.13±0.21^a	0.22±0.01^a	0.38±0.01^a
丰富环境组	3	3.83±1.65^a	13.82±1.86^a,b	67.15±3.64^a	158.70±1.58^a	87.23±1.35^a	56.24±2.96	0.24±0.02^a	0.41±0.00^a,b
F值		179.228	193.752	2994.473	13650.446	3508.858	134.914	333.841	196.000
P值		< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

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[15]	凌梦钰,杨一卓,刘帅,叶超群. 运动训练对脑缺血再灌注大鼠认知功能及前额皮层神经元核抗原和突触素1表达的效果[J]. 《中国康复理论与实践》, 2021, 27(11): 1272-1281.

丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响

Effects of Enriched Environment on Neurological Function and Glucose Metabolism in Ischemic Penumbra in Cerebral Ischemia-reperfusion Injury Rats

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