靶向线粒体治疗脊髓损伤的疗效：基于动物实验的系统评价

doi:10.3969/j.issn.1006-9771.2021.05.010

Abstract

Abstract: Objective

To systematically evaluate the effect of targeting mitochondria on spinal cord injury animal models, and provide experimental evidence.

Methods

Literatures about animal experiments of targeting mitochondria treatment for spinal cord injury were searched in PubMed, Web of Science, Web of Knowledge, CNKI and Wanfang database from establishment to February, 2021. Three researchers independently screened the literatures and extracted the data, and they were summarized by qualitative analysis.

Results

Eleven animal experimental studies were enrolled, including 548 animals with spinal cord injury. Six studies selected male or female Sprague-Dawley rats, and the rats in eight studies weighed 150~275 g. The animal models of spinal cord injury in all studies focused on T₉~T₁₁ contusive spinal cord injury, but there were differences in the use of spinal cord strikers and striking strength. The type, time, frequency, concentration and dosage of intervention drugs were all different. Due to the large heterogeneity of the included studies in animal species, animal models and outcome measures, qualitative analysis was conducted.

Conclusion

Targeting mitochondria for spinal cord injury in animals could promote the recovery of motor function, reduce the damaged spinal cord tissue and increase the remaining tissue, enhance the ability of anti-oxidation and anti-apoptosis, and enhance mitochondrial biogenesis. Limited by the number and quality of included studies, the above conclusions need to be verified by more high-quality studies.

Key words: spinal cord injury, mitochondria, dysfunction, treatment, animal experiment, systematic review

CLC Number:

R651.2

Li-qun WANG,Ri-zhao PANG,Jian-cheng LIU,An-ren ZHANG. Effect of Targeting Mitochondria on Spinal Cord Injury: A Systematic Review Based on Animal Experiments[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(5): 574-582.

Figures/Tables 6

作用机制	在脊髓损伤中的体现	相关药物
替代性生物能源	脊髓损伤后，由于氧化损伤会造成一些线粒体基质中可用的关键线粒体酶活性降低或失活，如丙酮酸脱氢酶，是一种产生乙酰辅酶A的关键酶，乙酰辅酶A是柠檬酸循环和生成ATP合成的电子供体所必需的生物能源，而脊髓损伤后丙酮酸脱氢酶活性的显著降低减少了柠檬酸循环中可用的乙酰辅酶A^[16]，因此需要引入其他可以用作能量产生的替代性生物能源	ALC
抗氧化	脊髓损伤后活性氧的形成增加，超出抗氧化剂系统的中和能力，促使强氧化剂过氧亚硝酸盐生成增加，诱导线粒体内一氧化氮合酶的活化，还可以触发细胞膜脂质过氧化，造成线粒体功能障碍。因此需要开发抗氧化剂靶向线粒体治疗脊髓损伤	α-生育酚、Neu2000、米诺环素、痿证中药方
抑制形成mPTP	脊髓损伤会造成线粒体内膜电位的破坏和mPTP的形成，从而导致细胞坏死和死亡，加剧损伤。抑制mPTP的形成促进线粒体功能	NIM811
抗凋亡	mPTP的开放导致线粒体外膜破裂，累积活性氧和许多凋亡相关蛋白(如细胞色素C、凋亡诱导因子、SMAC/Diablo)释放到细胞质中。这会导致多种形式的程序性细胞死亡，如凋亡	利拉鲁肽、痿证中药方
增强线粒体的生物发生	线粒体生物合成(mitochondrial biogenesis, MB)是指线粒体的修复、生长和分裂，涉及调控多种线粒体DNA编码基因的多种途径的复杂网络。受损的MB可能导致线粒体和细胞功能障碍。过氧化物酶体增殖物激活的受体γ共激活因子(peroxisomal proliferator γ coactivator, PGC-1α)是MB的主要调节剂^[17]。脊髓损伤后脊髓中PGC-1α表达降低，脊髓损伤后PGC-1α的维持和过表达则均减少神经元死亡并改善功能恢复^[18]。各种药物可以通过多种途径增强MB。5-羟色胺和β-肾上腺素能受体的激动作用可以激活蛋白激酶B、内皮型一氧化氮合酶或其他途径增强MB	LY344864、福莫特罗

纳入研究	动物种属	体质量(g)	n	脊髓损伤动物模型	干预措施
纳入研究	动物种属	体质量(g)	n	脊髓损伤动物模型	实验组	对照组
Patel等^[28](2010)	雌性Sprague-Dawley大鼠	200~250	132	T₁₀挫伤(Infinite Horizons脊髓打击器，力度200 kdyn)	ALC(术后15 min、30 min、60 min，腹腔注射300 mg/kg，干预3 h后处死)	生理盐水
沈娟等^[29](2019)	雌性Sprague-Dawley大鼠	200~240	64	T₁₀挫伤(HI-0400脊髓打击器，力度200 kdyn)	ALC(术后30 min内腹腔注射100 mg/kg、200 mg/kg、300 mg/kg，每天1次，连续7 d，之后每周1次，至术后第5周)	生理盐水
Cordero等^[30](2018)	雌性Sprague-Dawley大鼠	－	60	T₉~T₁₀挫伤(NYU脊髓打击器，12.5 mm高处释放10 g杆对脊髓进行重量冲击)	α-生育酚(术前8周，补充51 IU/g的α-生育酚在常规饮食中)	常规饮食
Springer等^[31](2010)	雌性Long-Evans大鼠	225~250	18	T₁₀挫伤(Infinite Horizons脊髓打击器，力度120 kdyn)	Neu2000(术后10 min腹腔注射单剂量50 mg/kg或在此基础上，再每24 h施用25 mg/kg，持续6 d)	生理盐水
Morsy等^[32](2010)	雄性Sprague-Dawley大鼠	150~200	50	脊髓缺血再灌注损伤(剖腹手术，在分叉上初夹住腹主动45 min，释放夹钳48 h再灌注)	α-生育酚(术后肌肉注射600 mg/kg，每周2次，持续6周)	安慰剂
Aras等(2015)^[33]	雄性Wistar大鼠	200~300	38	T₉挫伤(10 g重物5 cm高度垂直打击)	米诺环素(术后1 h和24 h经口服用3 mg/kg、30 mg/kg、90 mg/kg，然后处死)	空白对照和不同剂量相互对照
洪建仁等^[34](2020)	雄性Sprague-Dawley大鼠	200~250	36	T₁₀挫伤(Allen's法：将10 g重物5 cm高度垂直打击)	利拉鲁肽(术后即刻在硬膜内给药12 μg/100 g)	生理盐水
吕鹏等^[35](2021)	雄性Sprague-Dawley大鼠	225~275	30	T₁₀挫伤(改良Allen法：10 g打击锤放置从2.5 cm高度垂直打击)	痿证中药方(术后灌胃给药，按成人用量的1/60×大鼠体质量×10，每天1次，药量2 ml，连续 30 d)	空白对照
Springer等^[36](2018)	雌性Long-Evans大鼠	225~250	56	T₁₀挫伤(Infinite Horizons脊髓打击器，力度200 kdyn)	NIM811(术后1 h、12 h、24 h经口管饲单剂量10 mg/ml、20 mg/ml、40 mg/ml)	生理盐水和不同剂量相互对照
Simmons等^[37](2020)	雌性C57bl/6J小鼠	－	28	T₁₁挫伤(IH-0400打击器，力度200 kdyn)	LY344864(术后1 h腹腔注射，2 mg/kg，每天1次，持续21 d)	0.5%二甲基亚砜盐水
Scholpa等^[38](2019)	雌性C57Bl/6小鼠	－	36	T₁₁挫伤(IH-0400打击器，力度80 kdyn)	福莫特罗(术后8 h腹腔注射，每天0.1 mg/kg，持续48 d	1%二甲基亚砜盐水

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功能	相关评定方法或指标
运动功能	①BBB评分(Basso-Beattie-Bresnahan Locomotor Scale, BBB)^[19]；②Rivlin斜板实验^[20]；③Basso小鼠评分(Basso Mouse Scale, BMS)^[21]
组织病理学	①HE染色(观察脊髓组织形态学结构)；②铬花青染色^[22](eriochrome cyanine R)；③受损组织和残余组织体积^[23]
抗氧化功能	①丙二醛(malondialdehyde, MDA)；②超氧化物歧化酶(superoxide dismutase, SOD)；③谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)；④过氧化氢酶(catalase, CAT)
抗凋亡功能	①TUNEL染色；②凋亡相关蛋白和基因^[24]：Smac/Diablo、细胞色素C、FAS配体、肿瘤坏死因子-α、缺氧诱导因子-1α、P53、Bax和Bcl-2等
增强线粒体生物发生	①损伤部位的线粒体DNA和蛋白质含量；②线粒体生物发生相关蛋白^[25]：PGC-1α和ATP合酶β (ATP synthase β, ATPSynβ)；③线粒体总体生物能

结局指标	纳入研究	方法学限制	相关性	结果一致性	数据充分性	总体评价
运动功能	9项^{[29-32,34-38]}	9项研究中，2项存在选择、实施和失访偏倚，3项测量存在偏倚	9项研究纳入标准与拟解决问题基本一致	9项研究均报告运动功能改善	9个研究均进行定量描述，但使用的评定方法，数据的测量时间点和单位不一致，无法进行合并分析	高
脊髓组织病理学	8项^{[28-19,32,34-38]}	8项研究中，3项存在选择和实施偏倚，3项测量存在偏倚	8项研究纳入标准与拟解决问题基本一致	8项研究报告受损组织面积减少和残存组织面积增多，5项进行定量分析	8项研究中，5项研究进行定量描述，但使用的测量方法，数据的测量时间点和单位不一致，无法进行合并分析	高
抗氧化功能	6项^[29-33,35]	6项研究中，2项存在选择、实施、测量和报告偏倚	6项研究纳入标准与拟解决问题基本一致	6项研究报告抗氧化能力增强	6项研究进行定量描述，但具体功能指标，数据的测量时间点和单位不一致，无法进行合并分析	中
抗凋亡功能	2项^[29,34]	2项研究均存在选择、实施和测量偏倚	6项研究纳入标准与拟解决问题基本一致	2项研究报告抗凋亡能力增强	2项研究进行定量描述，但具体功能指标，数据的测量时间点和单位不一致，无法进行合并分析	低
增强线粒体生物发生	3项^[28,37-38]	3项研究中，1项存在选择、实施、测量和失访偏倚	6项研究纳入标准与拟解决问题基本一致	3项研究报告线粒体生物发生增强	3项研究进行定量描述，但具体功能指标，数据的测量时间点和单位不一致，无法进行合并分析	低

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Effect of Targeting Mitochondria on Spinal Cord Injury: A Systematic Review Based on Animal Experiments

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纳入研究	1	2	3	4	5	6	7	8	9	10	条目符合率
Patel等^[28](2010)	+	+	0	+	0	+	+	+	+	+	8/10
沈娟等^[29](2019)	+	+	0	+	0	0	0	+	+	+	6/10
Cordero等^[30](2018)	+	+	+	+	+	+	+	+	+	+	10/10
Springer等^[31](2010)	+	+	+	+	+	+	+	—	0	+	8/10
Morsy等^[32](2010)	+	+	+	+	+	+	+	+	+	+	10/10
Aras等(2015)^[33]	+	+	0	+	0	0	0	—	+	+	5/10
洪建仁等^[34](2020)	+	+	0	+	0	+	0	+	+	+	7/10
吕鹏等^[35](2021)	+	+	+	+	+	+	+	+	+	+	10/10
Springer等^[36](2018)	+	+	+	+	+	+	+	+	+	+	10/10
Simmons等^[37](2020)	+	+	+	+	+	0	0	—	+	+	7/10
Scholpa等^[38](2019)	+	+	+	+	+	+	+	+	+	+	10/10