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目的观察微管相关蛋白Tau在缺氧缺血性脑损伤(HIBD)新生大鼠脑组织中的表达变化,探讨HIBD早期Tau蛋白与细胞凋亡的关系。方法新生7日龄Wistar大鼠80只,随机分为2组:假手术组和HIBD组,每组各40只。HIBD组采用Rice方法建立新生大鼠HIBD模型。分别于手术后3、6、12、24、48 h取其脑组织切片,采用免疫组织化学方法检测其脑室周围白质区Tau的表达,末端脱氧核苷酸转移酶缺口标志(TUNEL)法检测其神经细胞凋亡。假手术组不行缺氧缺血处理,采用同样方法检测其Tau蛋白及神经细胞凋亡。结果假手术组脑室周围白质有少量Tau表达,且有少量凋亡细胞,各时间点无明显变化(Pa>0.05);HIBD组Tau的表达在6 h后开始增加,24 h达高峰,并持续至48 h,与假手术组比较差异均有统计学意义(Pa<0.05);细胞凋亡在6 h即开始增加,在12、24、48 h逐渐增加,与假手术组比较差异均有统计学意义(Pa<0.05);直线相关回归分析显示不同时间点HIBD组Tau蛋白表达与细胞凋亡呈显著正相关(Pa<0.05)。结论HIBD可造成新生大鼠脑室周围白质中神经纤维变性及Tau蛋白表达增加;Tau蛋白表达异常对促进神经细胞凋亡起重要作用。
Objective To observe the changes of microtubule-associated protein Tau in brain tissue of neonatal rats with hypoxic-ischemic brain damage (HIBD) and to explore the relationship between Tau protein and cell apoptosis in early stage of HIBD. Methods Eighty newborn 7-day-old Wistar rats were randomly divided into 2 groups: sham operation group and HIBD group, 40 rats in each group. HIBD group using Rice method to establish neonatal rat HIBD model. The brain tissue sections were taken at 3, 6, 12, 24, and 48 h after operation, respectively. The Tau expression in periventricular white matter was detected by immunohistochemistry. The TUNEL method was used to detect the Tau expression Nerve cell apoptosis. Sham-operation group was not hypoxic-ischemic treatment, using the same method to detect Tau protein and neuronal apoptosis. Results There was a small amount of Tau expression in the periventricular white matter and a small amount of apoptotic cells in the sham-operation group, with no significant changes at each time point (P> 0.05). The expression of Tau in HIBD group began to increase after 6 hours and peaked at 24 hours (P <0.05). The apoptosis rate began to increase at 6 h and gradually increased at 12 h, 24 h and 48 h, and the difference was statistically significant compared with the sham-operated group (P <0.05). Linear regression analysis showed that there was a significant positive correlation between Tau protein and apoptosis in HIBD group at different time points (P <0.05). Conclusions HIBD can cause neurofibrosis and Tau protein expression in periventricular white matter in neonatal rats. Abnormal expression of Tau plays an important role in promoting neuronal apoptosis.