衰老对大鼠脑出血后脑损伤的影响

doi:10.11817/j.issn.1673-9248.2021.06.003

目的

探讨不同年龄阶段脑出血后脑损伤的变化。

方法

取3月龄及22月龄SD大鼠，给予纹状体自体血注射构建脑出血模型，记录2组大鼠脑出血后7 d的生存率，脑出血第3天的神经功能损伤评分、脑含水量，并通过Nissl染色、TUNEL染色、Fluoro-Jade B染色检测脑出血后2组大鼠神经元损伤的差异。采用Kaplan-Meier生存分析生存率，并采用Log-rank（Mantel-Cox）检验比较2组的差异；采用独立样本t检验比较神经功能损伤评分、脑含水量及3种染色方法下神经元损伤的差异。

结果

22月龄脑出血大鼠的7 d生存率仅为20%，远低于3月龄组（80%），差异具有统计学意义（χ²=9.845，P=0.002）；22月龄脑出血大鼠3 d的神经损伤评分为（15.00±0.43）分，显著高于3月龄脑出血大鼠[（10.58±0.51）分]，差异具有统计学意义（t=6.611，P＜0.001）；22月龄脑出血大鼠3 d的脑含水量为[（85.10±0.60）]%，亦显著高于3月龄脑出血大鼠[（80.53±0.41）%]，差异具有统计学意义（t=6.335，P＜0.001）；Nissl、TUNEL及Fluoro-Jade B染色均发现22月龄脑出血大鼠的神经元损伤和凋亡较3月龄组显著增多，差异均具有统计学意义（P均＜0.01），提示22月龄大鼠脑出血神经元损伤更重。

结论

年龄的增大可加速脑出血所致的神经元损伤，导致神经功能恶化、水肿增多、耐受性下降。

关键词: 脑出血, 衰老, 脑损伤

Objectives

To compare the changes of brain injury after cerebral hemorrhage in different age stages in a rat model.

Methods

A model of cerebral hemorrhage was constructed using SD rats aged 3 months and 22 months with striatum autologous blood injection. The survival of 7 days after intracerebral hemorrhage was recorded and Kaplan-Meier survival analysis and Log-rank (Mantel-Cox) test were performed. The neurological deficient score and the content of brain edema were recorded on the third day of intracerebral hemorrhage. The difference of neuronal injury between the two groups was detected by Nissl, TUNEL and Fluoro-Jade B staining with Student's t-test for two groups' data comparison.

Results

7 days survival of 22-month-old rats with cerebral hemorrhage was 20%, which was significantly lower, than that of 3-month-old group (80%, χ²=9.845, P=0.002). Neurological deficient score of 22-month-old rats (mean±SD: 15.00±0.43) was higher than that of 3-month-old (10.58±0.5, t=6.611, P＜0.001), and the brain water content of 22-month-old rats (85.10±0.60)% was higher than that of 3-month-old [(80.53±0.41)%, t=6.335, P＜0.001]. Nissl, TUNEL and Fluoro-Jade B staining showed that the damage and apoptosis of neurons in 22-month-old cerebral hemorrhage rats increased significantly compared with 3-month-old group (P＜0.01), suggesting that cerebral hemorrhage induced neural damage was more serious in 22-month-old.

Conclusion

The increase of age can accelerate the neuronal injury caused by cerebral hemorrhage, which leads to the deterioration of neural function, the increase of edema and the decrease of injury tolerance.

Key words: Intracerebral hemorrhage, Aging, Cerebral injury

表1 实验大鼠体质量和血压数据比较（

x ˉ

±s）

大鼠月龄	样本量（只）	体质量（g）	收缩压（mmHg）	舒张压（mmHg）	脉压差（mmHg）
3月龄	48	348.3±42.3	115.6±1.34	84.23±1.20	31.40±1.05
22月龄	48	525.2±58.4	127.6±1.24	89.45±0.90	38.14±1.38
t值		12.185	7.126	3.494	3.888
P值		＜0.001	＜0.001	0.001	＜0.001

表1 实验大鼠体质量和血压数据比较（

x ˉ

±s）

大鼠月龄	样本量（只）	体质量（g）	收缩压（mmHg）	舒张压（mmHg）	脉压差（mmHg）
3月龄	48	348.3±42.3	115.6±1.34	84.23±1.20	31.40±1.05
22月龄	48	525.2±58.4	127.6±1.24	89.45±0.90	38.14±1.38
t值		12.185	7.126	3.494	3.888
P值		＜0.001	＜0.001	0.001	＜0.001

表2 3月龄和22月龄大鼠脑出血模型成模后不同时间点的神经功能评分比较（分，

x ˉ

±s）

大鼠月龄	样本量（只）	1 d	3 d	5 d	7 d
3月龄	12	5.75±0.43	10.58±0.51	6.33±0.47	2.67±0.31
22月龄	12	9.67±0.63	15.00±0.43	11.17±0.69	8.00±0.86
t值		5.131	6.611	5.779	5.825
P值		＜0.001	＜0.001	＜0.001	＜0.001

表2 3月龄和22月龄大鼠脑出血模型成模后不同时间点的神经功能评分比较（分，

x ˉ

±s）

大鼠月龄	样本量（只）	1 d	3 d	5 d	7 d
3月龄	12	5.75±0.43	10.58±0.51	6.33±0.47	2.67±0.31
22月龄	12	9.67±0.63	15.00±0.43	11.17±0.69	8.00±0.86
t值		5.131	6.611	5.779	5.825
P值		＜0.001	＜0.001	＜0.001	＜0.001

图1 不同月龄大鼠脑出血模型成模后7 d的Kaplan-Meier生存曲线

图2 不同月龄大鼠脑出血后第3天TUNEL染色结果。图a为脑出血后3 d，3月龄和22月龄大鼠出血侧纹状体及周边组织可见凋亡细胞（箭头所示）（n=5）；图b显示3月龄和22月龄大鼠脑出血后3 d脑组织凋亡细胞数量比较结果，差异具有统计学意义（n=5，t=21.21，^aP＜0.01）

图3 不同月龄大鼠脑出血后3 d Nissl染色结果。图a示脑出血后3 d，3月龄和22月龄大鼠脑出血对侧纹状体及周边组织可见大量Nissl小体（箭头所示），脑出血侧纹状体及周边组织Nissl小体数量显著减少；图b示3月龄和22月龄大鼠脑出血后3 d出血侧脑组织Nissl染色结果差异具有统计学意义（n=5，t=13.55，^aP＜0.01），对侧脑组织Nissl染色结果差异无统计学意义

图4 不同月龄大鼠脑出血后3 d FJB染色结果。图a示脑出血后3 d，3月龄和22月龄大鼠出血侧纹状体及周边组织可见变形神经元（箭头所示）；图b为3月龄和22月龄大鼠脑出血后3 d脑组织变性神经元数量比较结果，差异具有统计学意义（n=5，t=14.34，^aP＜0.01）

1	Tsai CF, Thomas B, Sudlow CL. Epidemiology of stroke and its subtypes in Chinese vs white populations: a systematic review [J]. Neurology, 2013, 81(3): 264-272.
2	Van Asch CJ, Luitse MJ, Rinkel GJ, et al. Incidence, case fatality, and functional outcome of intracerebral haemorrhage over time, according to age, sex, and ethnic origin: a systematic review and meta-analysis [J]. Lancet Neurol, 2010, 9(2): 167-176.
3	Liu M, Wu B, Wang W Z, et al. Stroke in China: epidemiology, prevention, and management strategies [J]. Lancet Neurol, 2007, 6(5): 456-464.
4	Jolink WM, Klijn CJ, Brouwers PJ, et al. Time trends in incidence, case fatality, and mortality of intracerebral hemorrhage [J]. Neurology, 2015, 85(15): 1318-1324.
5	Soo Y, Abrigo JM, Leung KT, et al. Risk of intracerebral haemorrhage in Chinese patients with atrial fibrillation on warfarin with cerebral microbleeds: the IPAAC-Warfarin study [J]. J Neurol Neurosurg Psychiatry, 2019, 90(4): 428-435.
6	Gong Y, Hua Y, Keep RF, et al. Intracerebral hemorrhage: effects of aging on brain edema and neurological deficits [J]. Stroke, 2004, 35(11): 2571-2575.
7	Wilkinson DA, Pandey AS, Thompson BG, et al. Injury mechanisms in acute intracerebral hemorrhage [J]. Neuropharmacology, 2018, 134(Pt B): 240-248.
8	Wan Y, Gao F, Ye F, et al. Effects of aging on hydrocephalus after intraventricular hemorrhage [J]. Fluids Barriers CNS, 2020, 17(1): 8.
9	Sumbria RK, Grigoryan MM, Vasilevko V, et al. Aging exacerbates development of cerebral microbleeds in a mouse model [J]. J Neuroinflammation, 2018, 15(1): 69.
10	Ni W, Okauchi M, Hatakeyama T, et al. Deferoxamine reduces intracerebral hemorrhage-induced white matter damage in aged rats [J]. Exp Neurol, 2015, 272: 128-134.
11	Chen J, Li Y, Wang L, et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats [J]. Stroke, 2001, 32(4): 1005-1011.
12	Xiong XY, Liu L, Wang FX, et al. Toll-like receptor 4/MyD88-mediated signaling of hepcidin expression causing brain iron accumulation, oxidative injury, and cognitive impairment after intracerebral hemorrhage [J]. Circulation, 2016, 134(14): 1025-1038.
13	Wang FX, Yang XL, Ma YS, et al. TRIF contributes to epileptogenesis in temporal lobe epilepsy during TLR4 activation [J]. Brain Behav Immun, 2018, 67: 65-76.
14	Tsai CF, Jeng JS, Anderson N, et al. Comparisons of risk factors for intracerebral hemorrhage versus ischemic stroke in chinese patients [J]. Neuroepidemiology, 2017, 48(1-2): 72-78.
15	An SJ, Kim TJ, Yoon BW. Epidemiology, risk factors, and clinical features of intracerebral hemorrhage: an update [J]. J Stroke, 2017, 19(1): 3-10.
16	Van Beijnum J, Lovelock CE, Cordonnier C, et al. Outcome after spontaneous and arteriovenous malformation-related intracerebral haemorrhage: population-based studies [J]. Brain, 2009, 132(Pt 2): 537-543.
17	郑嘉荣, 邓剑玲. 实验动物脑出血模型的研究进展 [J]. 医学综述, 2020, 26(5): 960-964.
18	Yuan JJ, Zhang Q, Gong CX, et al. Young plasma ameliorates aging-related acute brain injury after intracerebral hemorrhage [J]. Biosci Rep, 2019, 39(5): BSR20190537.
19	Lee JC, Cho GS, Choi BO, et al. Intracerebral hemorrhage-induced brain injury is aggravated in senescence-accelerated prone mice [J]. Stroke, 2006, 37(1): 216-222.
20	Gong Y, Xi GH, Keep RF, et al. Aging enhances intracerebral hemorrhage-induced brain injury in rats [J]. Acta Neurochir Suppl, 2005, 95: 425-427.
21	Zeng Z, Gong X, Hu Z. L-3-n-butylphthalide attenuates inflammation response and brain edema in rat intracerebral hemorrhage model [J]. Aging (Albany NY), 2020, 12(12): 11768-11780.
22	Dai S, Hua Y, Keep RF, et al. Minocycline attenuates brain injury and iron overload after intracerebral hemorrhage in aged female rats [J]. Neurobiol Dis, 2019, 126: 76-84.
23	王杰, 陈蔚翔, 夏敏, 等. 年龄对小鼠脑出血预后的影响 [J]. 中华神经外科杂志, 2018, 5(34): 524-529.
24	Wasserman JK, Yang H, Schlichter LC. Glial responses, neuron death and lesion resolution after intracerebral hemorrhage in young vs. aged rats [J]. Eur J Neurosci, 2008, 28(7): 1316-1328.
25	Lively S, Schlichter LC. SC1/hevin identifies early white matter injury after ischemia and intracerebral hemorrhage in young and aged rats [J]. J Neuropathol Exp Neurol, 2012, 71(6): 480-493.
26	Anqi X, Ruiqi C, Yanming R, et al. Neuroprotective potential of GDF11 in experimental intracerebral hemorrhage in elderly rats [J]. J Clin Neurosci, 2019, 63: 182-188.

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Effect of aging on brain injury in a rat model of cerebral hemorrhage

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Effect of aging on brain injury in a rat model of cerebral hemorrhage