Every-other-day fasting improves motor functional recovery of rats with clip-compression injury of the spinal cord

doi:10.3969/j.issn.2095-4344.0091

Abstract

Abstract:

BACKGROUND: Dietary intervention may be the most available, safe, economic and efficient approach to treat spinal cord injury.
OBJECTIVE: To observe the effects of dietary restriction in the form of every-other-day fasting (EODF) on the pathological changes and functional recovery of rats with clip-compression injury of the spinal cord.
METHODS: The models of spinal cord injury were established by a medical aneurysm clip in 36 Sprague-Dawley rats, and model rats were then randomized into four groups: group A with spinal cord injury and EODF, group B with spinal cord injury, group C with sham operation and EODF, and group D with sham operation. Food intake and body mass were observed. Motor functional recovery in rats was assessed by Basso, Beattie, Bresnahan scores at preoperative 1 day and postoperative 1 day, 1, 2, 4, 6, 8, 10, and 12 weeks. At 12 weeks after operation, the morphological changes of the spinal cord were observed through hematoxylin-eosin staining.
RESULTS AND CONCLUSION: The Basso, Beattie, Bresnahan score in the group B was decreased to the lowest at postoperative 1 day, and gradually increased with time. At 8-12 weeks after operation, the scores in the group A were significantly superior to those in the group B. At 12 weeks after operation, hematoxylin-eosin staining showed less lesion of the spinal cord in the group A than the group B. These results indicate that EODF can improve the motor functional of rats with spinal cord injury, and exerts effective protection on the injured spinal cord.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Spinal Cord Injuries, Intervention Studies, Caloric Restriction, Tissue Engineering

CLC Number:

R318

Sun Nian-yi1, 2, Xiong Xing-juan1, He Yu3, Wang Wen-chun2, Zhang An-ren2. Every-other-day fasting improves motor functional recovery of rats with clip-compression injury of the spinal cord[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(4): 564-569.

Figures/Tables 1

2.1 实验动物数量分析整个实验中有4只大鼠死亡，均按照实验步骤予以补充。实验最终纳入分析的大鼠数量是36只。 2.2 一般情况 4组大鼠均于术后2 h左右苏醒，脊髓损伤+隔日限食组、脊髓损伤组大鼠术后呈双足背着地、拖地而行的后肢瘫痪表现。假手术+隔日限食组、假手术组部分大鼠出现轻度步态异常，术后一两天完全恢复正常。所有大鼠手术切口愈合良好、无感染，手术缝线于术后2周左右自动脱落。术前各组大鼠每日进食量与体质量没有显著差异(图1)。术后4周当中，隔日限食组大鼠在进食日的进食量显著多于自由进食组(P < 0.01；图1 A)。其中，在第2，3周，脊髓损伤+隔日限食组大鼠进食日的进食量较脊髓损伤组大鼠相比增加了约90%(P < 0.01，P=0.028)，在第2-4周，假手术+隔日限食组大鼠进食日的进食量较假手术组大鼠相比增加了约75%(P < 0.01)。不过，尽管隔日限食组大鼠在进食当日的进食量对禁食期间的零摄入量进行了一定补偿，但是其总体摄食量较自由进食组减少(约10%)。术后，脊髓损伤+隔日限食组、脊髓损伤组、假手术+隔日限食组大鼠体质量呈现出1-3周的下降趋势后逐渐增加，脊髓损伤组和假手术+隔日限食组大鼠分别在术后8周和3周左右恢复至术前水平，在术后12周脊髓损伤+隔日限食组大鼠体质量仍未恢复至术前水平(图1B)。 2.3 运动功能评定结果术前各组大鼠BBB评分均为21分，评分差异均无显著性意义(P > 0.05)。脊髓损伤+隔日限食组、脊髓损伤组大鼠BBB评分在术后1 d降至最低，随时间逐渐增加，术后第8，10，12周，脊髓损伤+隔日限食组优于脊髓损伤组，组间差异有显著性意义(P < 0.05)；各时间点各组大鼠BBB评分见表1。 2.4 脊髓组织病理变化术后12周，脊髓损伤+隔日限食组(图2A)：视野中脊髓白质内可见空泡，可见脊髓神经纤维轴突萎缩、髓鞘肿胀，如红色箭头所示。脊髓损伤组(图2B)：视野中脊髓神经纤维中有许多明显可见的空泡，如黑色箭头所示；可见纤维细胞增生，如黄色箭头所示；可见有炎性细胞浸润，如绿色箭头所示。假手术+隔日限食组(图2C)：组织形态结构正常，未见明显病理改变。D组(图2D)：组织形态结构正常，未见明显病理改变。苏木精-伊红染色结果显示脊髓损伤+隔日限食组大鼠较脊髓损伤组大鼠脊髓损伤病变程度轻，隔日限食对大鼠脊髓损伤具有一定的保护作用。"

References

[1] Lee BB, Cripps RA, Fitzharris M,et al. The global map for traumatic spinal cord injury epidemiology: update 2011, global incidence rate.Spinal Cord.2014;52(2):110-116.

[2] 吴周睿,朱元贵,程黎明,等.脊髓损伤与修复的关键科学问题[J].中国科学基金,2013,3:137-153.

[3] Colman RJ, Anderson RM, Johnson SC, et al. Dietary restriction delays disease onset and mortality in rhesus monkeys. Science.2009;325(5937): 201-204.

[4] McCay CM, Maynard LA, Sperling G, et al. Retarded growth, lifespan, ultimate body size and age changes in the albino rat after feeding diets restricted in calories. J Nutr. 1939; 18(1):13.

[5] Martin B, Mattson MP, Maudsley S. Caloric restriction and intermittent fasting: two potential diets for successful brain aging. Ageing Res Rev. 2006;5(3):332-353.

[6] Meyer TE, Kovacs SJ, Ehsani AA, et al. Long-term caloric restriction ameliorates the decline in diastolic function in humans. J Am Coll Cardiol. 2006;47(2):398-402.

[7] Yu ZF, Mattson MP. Dietary restriction and 2-deoxyglucose administration reduce focal ischemic brain damage and improve behavioral outcome: evidence for a preconditioning mechanism. J Neuroscience Res.1999; 57:830-839.

[8] Vasudevan AR, Wu H, Xydakis AM, et al. Eotaxin and obesity. J Clin Endocrinol Metab.2006; 91:256-261.

[9] Colombo M, Kruhoeffer M, Gregersen S, et al. Energy restriction prevents the development of type 2 diabetes in Zucker diabetic fatty rats: coordinated patterns of gene expression for energy metabolism in insulin-sensitive tissues and pancreatic islets determined by oligonucleotide microarray analysis. Metabolism. 2006;55(1):43-52.

[10] Patel NV,Gordon MN,Connor KE,et al.Caloric restriction attenuates Abeta-deposition in Alzheimer transgenic models. Neurobiol Aging. 2005;26(7):995-1000.

[11] Qin W,Yang T,Ho L,et al. Neuronal SIRT1 activation as a novel mechanism underlying the prevention of Alzheimer disease amyloid neuropathology by calorie restriction. J Biological Chemistry.2006; 281:21745-21754.

[12] Wang J,Ho L,Qin W,et al.Caloric restriction attenuates beta-amyloid neuropathology in a mouse model of Alzheimer's disease.J Faseb.2005; 19:659-661.

[13] 侯亚萍. 热量限制及其对阿尔茨海默等退行性疾病的可能作用[J]. 解剖学研究, 2010, 32:52-58.

[14] Varady KA, Hellerstein MK. Alternate-day fasting and chronic disease prevention: a review of human and animal trials. Am J Clin Nutr.2007;86:7-13.

[15] Anson RM, Guo Z, de Cabo R, et al. Intermittent fasting dissociates beneficial effects of dietary restriction on glucose metabolism and neuronal resistance to injury from calorie intake. Proc Natl Acad Sci USA.2003;100:6216-6220.

[16] Bsso DM,Beattie MS,Bresnahan JC.A sensitive and reliable locomotor rating scale for open field testing in rats.J Neurotrauma.1995;12(01):1-21.

[17] Hall ED, Springer JE. Neuroprotection and acute spinal cord injury: a reappraisal. NeuroRx .2004;(1):80-100.

[18] Dumont RJ, Okonkwo DO, Verma S, et al. Acute spinal cord injury, part I: pathophysiologic mechanisms. Clin Neuropharmacol.2001;24(5): 254-264.

[19] Carlson GD, Gorden C. Current developments in spinal cord injury research. Spine J.2002;2(2):116-128.

[20] 席悦.利用微阵列芯片技术探究脊髓损伤的分子机制[J]. 中国组织工程研究,2013,17(24):4529-4538.

[21] Ramer LM, Ramer MS, Bradbury EJ. Restoring function after spinal cord injury: towards clinical translation of experimental strategies. The Lancet Neurology. 2014;13(12):1241-1256.

[22] Colman RJ, Anderson RM, Johnson SC, et al. Caloric restriction delays disease onset and mortality in rhesus monkeys. Science.2009;325: 201-204.

[23] Vermeij WP, Dollé ME, Reiling E, et al. Restricted diet delays accelerated ageing and genomic stress in DNA-repair-deficient mice. Nature.2016;537(7620): 427-431.

[24] Mattison JA, Colman RJ, Beasley TM, et al. Caloric restriction improves health and survival of rhesus monkeys. Nat Commun. 2017;8:14063.

[25] Cheng CW, Villani V, Buono R, et al. Fasting-Mimicking Diet Promotes Ngn3-Driven β-Cell Regeneration to Reverse Diabetes.Cell.2017;168(5):775-788.

[26] Michalsen A, Li C.Fasting Therapy for Treating and Preventing Disease Current State of Evidence. Forsch Komplementmed. 2013;20(6):444-453.

[27] Khaleghi Ghadiri M, Tutam Y, Wassmann H,et al.Periodic fasting alters neuronal excitability in rat neocortical and hippocampal tissues. Neurobiol Dis. 2009;36(2):384-392.

[28] Fann DY, Santro T, Manzanero S, et al. Intermittent fasting attenuates inflammasome activity in ischemic stroke. Exp Neurol. 2014;257:114-119.

[29] Manzanero S, Erion JR, Santro T, et al.Intermittent fasting attenuates increases in neurogenesis after ischemia and reperfusion and improves recovery. J Cereb Blood Flow Metab. 2014;34(5):897-905 .

[30] Laurie M. Davis, James R. Pauly, Ryan D. Readnower, et al. Fasting Is Neuroprotective Following Traumatic Brain Injury. J Neurosci Res.2008;86:1812-1822.

[31] Arumugam TV, Phillips TM, Cheng A, et al. Age and energy intake interact to modify cell stress pathways and stroke outcome. Ann Neurol.2010;67(1): 41-52.

[32] Jeong JH, Yu KS, Bak DH, et al. Intermittent fasting is neuroprotective in focal cerebral ischemia by minimizing autophagic flux disturbance and inhibiting apoptosis. Exp Ther Med.2016;12(5): 3021-3028.

[33] Jeong MA, Plunet W, Streijger F, et al. Intermittent fasting improves functional recovery after rat thoracic contusion spinal cord injury. J Neurotrauma.2011;28: 479-492.

[34] 刘捷. 饮食干预疗法对急性脊髓损伤治疗作用的研究[J]. 中国骨科临床与基础研究杂志, 2012, 4(1): 75.

[35] Plunet WT, Streijger F, Lam CK, et al. Dietary restriction started after spinal cord injury improves functional recovery. Exp Neurol.2008;213(1): 28-35.

[36] Plunet WT,Lam CK,Lee JH,et al.Prophylactic dietary restriction may promote functional recovery and increase lifespan after spinal cord injury. Ann NY Acad Sci.2010;1198 Suppl 1: E1-11.

[37] 刘建成,王文春,罗少杰,等. 隔日限食疗法对大鼠急性脊髓损伤后血清TNF-α的影响[J]. 康复学报,2015,25(4):14-17.

[38] 刘建成. 隔日限食疗法对大鼠急性脊髓损伤后神经保护作用的机制研究[D]. 成都:成都中医药大学,2015.