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目的 :观察核因子-κB p50(nuclear factor-κB subunit p50,NF-κB p50)特异性抑制剂吡咯烷二硫代氨基甲酸(pyrrolidine dithiocarbamate,PDTC)抑制NF-κB p50后,炎症因子TNF-α及相关康复蛋白Reelin、突触素、Lingo-1表达的变化,进一步观察大鼠神经功能评分、海马神经细胞形态的变化。方法:本实验的研究对象为雄性健康SD大鼠,随机分为3组:假手术组(S组,n=6)、缺血组(I组,n=18)、PDTC干预组(P组,n=18)。脑缺血模型采用永久性MCAO法。P组的大鼠在造模后30 min进行腹腔注射PDTC(100 mg/kg),每天1次。S组大鼠于第3天处死,I组和P组大鼠分别于1、3、7 d处死。用Longa评分对3组大鼠进行神经功能损害程度进行评估,用Western blot检测大鼠缺血海马NF-κBp50、TNF-α、Reelin、突触素蛋白的表达,用HE染色方法观察3组大鼠海马神经元细胞的形态,用免疫组化法检测Lingo-1在脑组织内的表达情况。结果:1P组及I组大鼠Longa评分较S组均有所升高,术后第1天和第3天P组较I组Longa评分降低,但无统计学意义(P>0.05);在术后第7天,P组较I组显著降低(P<0.05)。2I组大鼠海马神经细胞出现异常,而P组大鼠中,海马神经元细胞的结构基本保持正常。3术后1、3天,I组和P组大鼠缺血海马中NF-κB p50较S组升高(P<0.05),但P组NF-κB p50表达较I组降低(P<0.05);术后7天,3组变化无统计学意义。4术后1、3、7天,I组和P组大鼠TNF-α表达较S组升高,但P组TNF-α较I组降低(P<0.05);5术后第1、3天,P组大鼠缺血海马中突触素和Reelin蛋白的表达明显高于I组(P<0.05),第7天两组差别无统计学意义(P>0.05)。S组大鼠脑组织内可见少量Lingo-1阳性细胞表达,P组大鼠脑组织内Lingo-1阳性细胞数在第7天时明显低于I组。结论:PDTC可改善神经功能,维持海马神经元正常形态,这可能与其抑制NF-κB p50表达,减少TNF-α等相关炎症因子生成及调节Reelin、突触素及Lingo-1表达有关。
OBJECTIVE: To observe the effects of pyrrolidine dithiocarbamate (PDTC), a specific inhibitor of nuclear factor-κB subunit p50 (NF-κB p50), on the expression of inflammatory cytokines TNF-α And reelin, synaptophysin and Lingo-1 expression in rats were observed. The changes of neurological function score and hippocampal neuron morphology were further observed. Methods: The male Sprague-Dawley rats were randomly divided into 3 groups: sham operation group (n = 6), ischemia group (n = 18), PDTC intervention group , n = 18). Cerebral ischemia model using permanent MCAO method. Rats in group P were given intraperitoneal injection of PDTC (100 mg / kg) 30 min after modeling, once daily. Rats in group S were sacrificed on day 3, and rats in group I and group P were sacrificed on days 1,3,7. Longa score was used to evaluate the degree of neurological dysfunction in rats. Western blot was used to detect the expression of NF-κBp50, TNF-α, Reelin and synaptophysin in hippocampus of rats. Immunohistochemistry was used to detect the expression of Lingo-1 in brain tissue of hippocampal neurons. Results: The Longa score of rats in 1P group and I group was higher than that of S group. The Longa score of P group was lower than that of I group on the first day and the third day after operation (P> 0.05) On the 7th day after operation, the P group was significantly lower than the I group (P <0.05). The neurons in hippocampus of 2I group were abnormal, but the structure of neurons in hippocampus of P group rats remained normal. (P <0.05) .3 The expression of NF-κB p50 in hippocampus of I and P groups was significantly lower than that of S group (P <0.05) ); 7 days after operation, there was no significant difference in the three groups. The levels of TNF-α in group I and group P were significantly higher than those in group S (P <0.05), but the level of TNF-α in group P was lower than that in group I (P <0.05) Day, the expression of synaptophysin and Reelin protein in hippocampus of group P was significantly higher than that of group I (P <0.05). There was no significant difference between the two groups on the seventh day (P> 0.05). A small amount of Lingo-1 positive cells were observed in the brain tissue of S group. The number of Lingo-1 positive cells in brain tissue of P group was significantly lower than that of I group on the 7th day. CONCLUSION: PDTC can improve neurological function and maintain the normal morphology of hippocampal neurons, which may be related to its inhibition of NF-κB p50 expression, reduction of TNF-α-related inflammatory cytokines and regulation of Reelin, Synaptophysin and Lingo-1.