| 钟超超,徐兴国,苗海航,等.亚低温对缺血性脑卒中小鼠血脑屏障损伤保护作用[J].安徽医药,2023,27(2):337-340. |
| 亚低温对缺血性脑卒中小鼠血脑屏障损伤保护作用 |
| Protective effect of subhypothermia on blood.brain barrier injury in mice with ischemic stroke |
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| DOI：10.3969/j.issn.1009-6469.2023.02.028 |
| 中文关键词: 低温，人工 脑梗死 缺血性脑卒中 血脑屏障 |
| 英文关键词: Hypothermia,induced Brain infarction Ischemic stroke Blood.brain barrier |
| 基金项目:南通市科技发展计划项目( MS12020026) |
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| 摘要点击次数: 734 |
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| 中文摘要: |
| 目的探讨亚低温对缺血性脑卒中小鼠血脑屏障的保护作用，寻找亚低温发挥保护作用的机制。方法选取由南通大学实验动物中心提供的 SPF级别雄性 C57/BL6小鼠 54只，分为假手术组( n=18)和卒中组( n=36)，卒中组分为亚低温组( n= 18)和常温组( n=18)。卒中组在行大脑中动脉阻塞手术后对亚低温组小鼠予以亚低温处理，在口腔温度监测下维持小鼠温度为( 34±0.5)℃，其他组术后均置于保温毯上正常复温。术后每日对各组神经功能进行评分，同时使用错步时间检测卒中后小鼠行为运动功能。于术后第 3天处死小鼠，取梗死脑区行蛋白质印迹法( Western blotting)检测血脑屏障黏附分子紧密连接蛋(ZO-1)紧密连接跨膜蛋白(Claudin-5)的蛋白表达以判断血脑屏障损伤的程度。结果假手术组、亚低温组、常温组小鼠存活白率分别为，100%(18/18)，94.44%(17/18)，88.89%(16/18)。三组存活率比较，差异无统计学意义( χ2=53.72，P=0.122)。卒中组小鼠术后第 1、2、3天神经功能评分［(2.8±0.2)、(2.6 ± 0.3)、(2.4±0.3)分］明显高于假手术组［( 0.6 ± 0.04)、(0.5 ± 0.03)、(0.4 ± 0.01)分］，差异有统计学意义( t=2.94，P<0.001；t=2.86，P<0.001；t=2.98，P<0.001)；卒中组中，常温组小鼠术后第 1、2、3天神经功能评分均明显高于亚低温组［(2.0±0.3)、(1.9±0.2)、(1.8±0.2)分］差异有统计学意义。卒中组小鼠术后第 1、2、3天前后肢的平均错步比例( 26.8%前、32%后；29.0%前、31.1%后；31.8%前、33.1%后)均，高于假手术组(均为 3%前、3%后)差异有统计学意义( χ2=29.05，P=0.002；χ2=28.76，P=0.003；χ2=26.42，P=0.005)且常温组小鼠术后第 1、2、3天前后肢平均的比例高于亚低温组错步，(16.6%前16.9%后；18.6%前、17.9%后；20.8%前、19.9%后)有统计学意义( χ2=27.35，P=0.004；χ2=26.59，P=0.004；χ2=24.12，P=差异，0.007)活的小鼠，脑卒中可以显著降低小鼠脑脑组织中血脑屏障黏附分子 ZO-1，Claudin-5的蛋白表达( t=2.54，P=0.001；t=2.46，P=0.003)，亚低温处理组小鼠脑组织血脑屏障黏附分子 ZO-1，Claudin-5的蛋白表达比常温处理组显著增高( t=2.48，P=0.003；t=1.68，P=0.043)。结论脑卒中术后予以 6h的亚低温处理可降低卒中后小鼠神经功能评分，改善术后短期运动行为功能，减少血脑屏障的破坏。 |
| 英文摘要: |
| Objective To investigate the protective effect of subhypothermia on blood.brain barrier injury in mice with ischemicstroke, and to explore the mechanism of the protective effect of subhypothermia. Methods A total of 54 SPF-grade male C57/BL6 mice provided by the Experimental Animal Center of Nantong University were selected and divided into a sham-operated group (n=18) and a stroke group (n=36), and the stroke group was divided into a subhypothermia group (n=18) and a normothermic group (n=18). Inthe stroke group, mice in the subcritical group were treated with subcritical temperature after middle cerebral artery obstruction sur-gery, and the temperature of the mice was maintained at (34±0.5)℃ under oral temperature monitoring, while all other groups wereplaced on a thermal blanket for normal rewarming after surgery. The neurological function of each group was scored daily after surgery,and the poststroke behavioral-motor function of the mice was also measured using staggered time. The mice were sacrificed on the thirdday after surgery, and the protein expression of ZO-1 and Claudin-5 was measured by western blotting to determine the extent of blood.brain barrier damage. Results The survival rates of the sham group, subhypothermia group and normothermic groups of mice were100% (18/18), 94.44% (17/18), and 88.89% (16/18), respectively. There was no significant difference in the survival rate among thethree groups (χ2=53.72, P=0.122). The neurological function scores on postoperative days 1, 2, and 3 were significantly higher in thestroke group [(2.8±0.2), (2.6±0.3), (2.4±0.3)] than in the sham group [(0.6±0.04), (0.5±0.03), and (0.4±0.01)], with statistically signifi- cant differences (t=2.94,P<0.001;t=2.86,P<0.001;t=2.98,P<0.001). In the stroke group, the neurological function scores on postopera-tive days 1, 2 and 3 were significantly higher in the normothermic group than in the subhypothermia group [(2.0±0.3), (1.9±0.2), and(1.8±0.2)], and the differences were statistically significant. In the stroke group, the average proportion of hind limb missteps before the1st, 2nd, and 3rd postoperative days was higher in the mice (26.8% forelimb and 32% posterior limb; 29.0% forelimb and 31.1% poste-rior limb; 31.8% forelimb and 33.1% posterior limb) than in the sham-operated group (both forelimb and posterior limb 3%), and the differences were statistically significant (χ2=29.05,P=0.002; χ2= 28.76, P=0.003; χ2=26.42, P=0.005). The average proportion of hindlimb missteps before the 1st, 2nd, and 3rd postoperative days were higher in the normothermic group than in the sham-operated group(16.6% forelimb and 16.9% posterior limb; 18.6% forelimb and 17.9% posterior limb; 20.8% forelimb and 19.9% posterior limb), andthe difference was statistically significant (χ2=27.35, P=0.004; χ2=26.59, P=0.004; χ2= 24.12, P=0.007). In surviving mice, stroke sig-nificantly reduced the protein expression of ZO-1 and Claudin-5 in the brain tissue of mice with stroke (t=2.54, P=0.001; t=2.46, P= 0.003), and the protein expression of ZO-1 and Claudin-5 in the brain tissue of mice treated with subhypothermia was significantly high-er than that in the normothermic treatment group (t=2.48, P=0.003; t=1.68, P=0.043).Conclusion Subhypothermia treatment for 6 h after stroke surgery reduced the neurological scores, improved the short-term postoperative motor behavioral function, and reduced the damage to the blood.brain barrier. |
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