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目的研究四氯化碳(CCl4)对大鼠脑海马CA1区神经元低亲和力神经生长因子受体p75的表达和凋亡的影响。方法将24只健康雄性Wistar大鼠随机分为对照组和CCl4组。对照组每周一、四背部皮下注射橄榄油(0.12ml/100g),CCl4组分别在每周一或/和周四背部皮下注射60%CCl4-橄榄油溶液(0.3ml/100g)1次(1d组)2、次(1周组)和4次(2周组),实验结束时处死大鼠。取脑后沿正中矢状面切开,右侧半脑石蜡切片,行Nissl染色,观察海马CA1区神经元的形态学改变;行p75、Bax免疫组织化学染色和Western blotting分析,检测海马CA1区神经元p75、Bax的表达,行原位缺口末端标记法(TUNEL)观察海马CA1区神经元的凋亡。结果Nissl染色显示对照组CA1区神经元染色较深,细胞数目较多,排列整齐,神经元内可见大量尼氏体;CCl4组CA1区神经元排列较对照组紊乱,部分锥体细胞体积缩小,核固缩,呈三角形,胞浆内尼氏体数目减少或消失。对照组海马CA1区可见少量表达p75和Bax的阳性细胞;CCl4组海马CA1区表达p75和Bax的阳性细胞数较对照组明显增多,以1周CCl4组增多最为明显;Western blotting结果与免疫组织化学染色结果一致。对照组海马CA1区未见TUNEL阳性细胞;CCl4组海马CA1区可见大量细胞核呈棕黄色着色的TUNEL阳性细胞,以1周CCl4组的阳性细胞数最多。结论注射CCl4后,大鼠脑海马CA1区p75的表达明显增强,Bax的表达也相应增强,凋亡的神经元明显增多。
Objective To investigate the effect of carbon tetrachloride (CCl4) on the expression and apoptosis of low affinity nerve growth factor receptor p75 in rat hippocampal CA1 neurons. Methods Twenty-four healthy male Wistar rats were randomly divided into control group and CCl4 group. The control group was injected with olive oil (0.12ml / 100g) subcutaneously on the first and fourth back, and CCl4 subcutaneously injected with 60% CCl4-olive oil solution (0.3ml / 100g) ) 2 times (1 week group) and 4 times (2 weeks group), rats were sacrificed at the end of the experiment. The brain was cut along the median sagittal plane and the right hemisphere paraffin sections were taken for Nissl staining. Morphological changes of hippocampal CA1 neurons were observed. Immunohistochemical staining and Western blotting were used to detect the hippocampal CA1 region The expression of p75 and Bax in neurons and the apoptosis of hippocampal CA1 neurons were observed by TUNEL. Results Nissl staining showed that neurons in CA1 area of the control group were stained darker with more cells and neatly arranged, and a large number of Nissl bodies could be seen in the neurons. The arrangement of neurons in CA1 area of CCl4 group was disordered compared with the control group, Nuclear pyknosis, triangular, intracytoplasmic Niger reduce or disappear. The number of positive cells expressing p75 and Bax in CA1 area of hippocampus in CCl4 group was significantly higher than that in control group, and the most significant increase was in CCl4 group of 1 week. Western blotting results were compared with immunohistochemistry The same staining results. There were no TUNEL positive cells in CA1 area of hippocampus in control group. A large number of TUNEL-positive cells were observed in CA1 area of CCl4 hippocampus, and the number of positive cells in CCl4 group was the highest at 1 week. Conclusion After injection of CCl4, the expression of p75 in hippocampal CA1 region of rat hippocampus significantly increased, and the expression of Bax increased correspondingly. The number of apoptotic neurons increased significantly.