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目的研究亚低温对新生大鼠缺氧缺血性脑损伤脑红蛋白表达的影响,探讨亚低温的脑保护作用机制。方法随机将出生7d的Wistar大鼠110只分为3组:假手术组10只,常温脑缺氧缺血组50只,亚低温脑缺氧缺血组50只。常温组和亚低温组再根据动物模型建立后1、5、15、30、60min5个时间点随机分为5个亚组,每个亚组10只大鼠,动物模型通过结扎双侧颈总动脉建立。常温组直肠温度控制在36~37℃,亚低温组直肠温度控制在32~33℃。所有动物均断头取脑,通过RT-PCR及免疫组化的方法检测脑红蛋白的表达。结果RT-PCR结果显示亚低温组在1、5、15、30min脑红蛋白相对表达水平(A:1.10±0.05、0.96±0.06、0.79±0.06、0.67±0.08)均高于常温组(A:0.98±0.07、0.81±0.11、0.53±0.08、0.36±0.04)有统计学意义(P<0.05),免疫组化显示亚低温组在5、15、30min脑红蛋白的积分光密度值IOD[(23.26±2.83)、(32.06±2.58)、(19.39±3.16)×105]均较常温组明显增高IOD[(19.16±2.16),(25.15±4.05),(13.78±2.40)×105],经比较差异有统计学意义(P<0.05)。结论亚低温增加脑红蛋白的表达可能是亚低温脑保护机制之一。
Objective To investigate the effects of mild hypothermia on the expression of neuroglobin in neonatal rats with hypoxic-ischemic brain damage (HIBD) and to explore the mechanism of hypothermia. Methods One hundred and ten Wistar rats were randomly divided into three groups: sham operation group (n = 50), hypothermia hypoxic ischemic group (n = 50) and hypothermia hypoxic ischemic group (n = 50). The normal temperature group and the mild hypothermia group were divided into 5 subgroups at 5, 15, 30 and 60 min after establishment of the animal model at each time point, and each subgroup was given 10 rats. The animal model was established by ligating bilateral common carotid arteries set up. The rectal temperature was controlled at 36 ~ 37 ℃ in the normal temperature group and 32 ~ 33 ℃ in the mild hypothermia group. All animals were decapitated and brains were removed. The expression of neuroglobin was detected by RT-PCR and immunohistochemistry. Results RT-PCR results showed that the relative expression levels of neuroglobin at 1, 5, 15, 30 min in mild hypothermia group were higher than those in normal temperature group (A: 1.10 ± 0.05, 0.96 ± 0.06, 0.79 ± 0.06, 0.67 ± 0.08) 0.98 ± 0.07,0.81 ± 0.11,0.53 ± 0.08,0.36 ± 0.04) (P <0.05), immunohistochemistry showed that the integral optical density value of IOD at 5, 15 and 30 min in mild hypothermia group [ 23.16 ± 2.83), (32.06 ± 2.58) and (19.39 ± 3.16) × 105, respectively. Compared with the normal temperature group, the IOD was significantly higher (19.16 ± 2.16, (25.15 ± 4.05), (13.78 ± 2.40) × 105) The difference was statistically significant (P <0.05). Conclusion Mild hypothermia increases the expression of neuroglobin may be one of the mechanisms of hypothermia brain protection.