TLR4及NF-kB在脑出血大鼠血肿周围组织的表达

The Expression and Significance of TLR4,NF-Kb in Hamatoma Tissues of Rats with Cerebral Hemorrhage

目的:探讨核因子-k B(NF-k B)和Toll样受体4(TLR4)在大鼠脑出血(ICH)后继发炎症损伤中的作用。方法:40只SD雄性大鼠随机均分为模型组和假手术组,模型组大鼠于前囟旁1 cm处颅骨钻孔并缓慢注入自体动脉血50μL建立ICH模型,假手术组注入等量生理盐水;两组大鼠又分为12、24、72 h和第7天组(每组5只),于相应时间点对大鼠进行神经功能障碍评分后,取大鼠部分脑组织切片,HE染色和尼氏染色观察大鼠脑组织病理改变,免疫组化染色观察TLR4、NF-k B阳性神经元,;同时取脑组织匀浆,采用Western blot检测血肿周围组织TLR4、NF-k B蛋白表达水平。结果:模型组各时点神经行为学评分、血肿周围组织未受损神经元数均明显低于假手术组(P<0.05);模型组72 h时神经行为学评分和血肿周围组织未受损神经元数少于模型组其他时间点(P<0.05);模型组各时点血肿周围组织TLR4、NF-k B蛋白阳性细胞数及蛋白表达量明显高于假手术组(P<0.05);模型组72 h时基底神经节TLR4、NF-k B蛋白表达阳性细胞数及表达量明显高于其他模型组(P<0.05)。结论:大鼠脑出血后,TLR4、NF-k B表达上调可能加重脑组织继发性损伤。

Objective： To investigate the expression and significance of nuclear factor-μB （NF-μB） and Toll like receptor 4 （TLR4） in rats with inflammation injury after intracerebral hemorrhage （ICH）. Methods： 40 male SD rats were randomly divided into two groups： sham operation group and model group （n = 20） ; rats in model group were established ICH rat model （performed I cm trepana- tion on anterior fontanelle and slowly injected 50 μL autologous arterial blood） ; at the same time, sham operation group and model group were divided into 12 h, 24 h, 72 h and 7 d groups by time point （5 in each group） ; used Garcia method to evaluate the neurological dysfunction, used HE stai- ning and Nissl staining to observe rat brain tissue pathological changes, used immunohistochemical staining to observe TLR4 and NF-kB NOS positive neurons, and Western blot to detect the protein ex- pression level of TLR4 and NF-κB protein in hematoma surrounding tissue. Results： Neuroethology score and the number of uninjured neurons surrounding the hematoma of model group at different time points were significantly lower than that of sham operated group （P 〈 0.05 ） ; at 72 h, neurobehavioral score of model group and the number of uninjured neurons in the hematoma surrounding tissues were less than other time points of model group （P 〈 0.05 ） ; the number of TLR4 and NF-κB protein posi tive cells and protein expression in the tissues around the hematoma were significantly higher than those in the sham operation group at different time points （ P 〈 0.05 ） ; at 72 h in the model group, number of positive cells and the expression of TLR4 and NF-kB protein in basal ganglia were significantly high- er than other time points of model group （ P 〈 0.05 ）. Conclusion ： After cerebral hemorrhage in rats, the expression up regulation of TLR4 and NF-κB may aggravate the secondary injury of brain tissue around hematoma.