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目的研究丁苯酞对大鼠急性重度CO中毒后脑组织中半胱氨酸蛋白-3(Caspase-3)和半胱氨酸蛋白酶-9(Caspase-9)表达的影响。方法通过数字表法将大鼠随机分为3组:正常组、模型组和实验组(均n=10)。用CO染毒法制作大鼠急性重度CO中毒模型。实验组大鼠给予丁苯酞80 mg·kg~(-1),灌胃1次/天,连续7 d,正常组和模型组给予等量的0.9%NaCl。在7 d后,用免疫组化染色法观测Caspase-3和Caspase-9阳性细胞数,用反转录酶-聚合酶链锁反应(RT-PCR)法及蛋白质印迹法检测Caspase-3和Caspase-9mRNA和蛋白的表达水平。结果正常组大鼠脑组织中Caspase-3和Caspase-9的阳性细胞数分别为4.98±1.34,8.65±1.89,mRNA表达水平分别为0.19±0.02,0.18±0.03,蛋白表达水平分别为0.09±0.01,0.08±0.01;模型组大鼠脑组织中Caspase-3和Caspase-9阳性细胞数分别为29.43±4.53,40.45±5.23,mRNA表达水平分别为0.47±0.05,0.75±0.05,蛋白表达水平分别为0.78±0.05,0.86±0.07。与正常组对比,模型组的Caspase-3和Caspase-9mRNA和蛋白表达水平显著增高,差异均有统计学意义(均P<0.05)。实验组大鼠脑组织中的Caspase-3和Caspase-9阳性细胞数分别为22.26±3.12,25.09±4.25,mRNA表达水平分别为0.35±0.05,0.41±0.05,蛋白表达水平分别为0.37±0.02,0.45±0.02,与模型组对比,Caspase-3和Caspase-9 mRNA和蛋白表达水平较明显下调,差异均有统计学意义(均P<0.05)。结论丁苯酞可通过下调Caspase-3和Caspase-9的表达水平来抑制神经细胞凋亡进程,从而减轻CO对大鼠脑神经的损害。
Objective To investigate the effects of butylphthalide on the expression of Caspase-3 and Caspase-9 in rats with acute severe CO poisoning. Methods The rats were randomly divided into 3 groups: normal group, model group and experimental group (n = 10). Acute Severe CO Poisoning Model in Rats Treated with CO. Rats in the experimental group were given butylphthalide (80 mg · kg -1) once a day for 7 days. The rats in the normal group and model group were given the same amount of 0.9% NaCl. After 7 days, the number of Caspase-3 and Caspase-9 positive cells was observed by immunohistochemical staining. Caspase-3 and Caspase-3 were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting -9 mRNA and protein expression levels. Results The positive cells of Caspase-3 and Caspase-9 in the normal group were 4.98 ± 1.34 and 8.65 ± 1.89, the mRNA expression levels were 0.19 ± 0.02 and 0.18 ± 0.03 respectively, the protein expression levels were 0.09 ± 0.01 , 0.08 ± 0.01 respectively. The positive cells of Caspase-3 and Caspase-9 in the model group were 29.43 ± 4.53 and 40.45 ± 5.23 respectively, the mRNA expression levels were 0.47 ± 0.05 and 0.75 ± 0.05 respectively 0.78 ± 0.05, 0.86 ± 0.07. Compared with the normal group, the expression of Caspase-3 and Caspase-9 mRNA and protein in the model group were significantly increased, the differences were statistically significant (all P <0.05). The number of Caspase-3 and Caspase-9 positive cells in the experimental group were respectively 22.26 ± 3.12 and 25.09 ± 4.25, the mRNA expression levels were 0.35 ± 0.05 and 0.41 ± 0.05 respectively, the protein expression levels were 0.37 ± 0.02, 0.45 ± 0.02. Compared with the model group, the mRNA and protein expressions of Caspase-3 and Caspase-9 were significantly down-regulated (all P <0.05). Butylphthalide can inhibit the neuronal apoptosis process by down-regulating the expression of Caspase-3 and Caspase-9, so as to reduce the damage of CO to the cranial nerves in rats.