脑舒明方对局灶性脑缺血大鼠NF-κB,TNF-α和IL-1β表达的影响

Effect of Naoshuming decoction on expression of NF-κB, TNF-α, and IL-1β in focal cerebral ischemia rats

目的:探讨脑舒明方对脑缺血大鼠的影响。方法:采用大脑中动脉阻塞法复制大鼠中脑动脉闭塞(middle cerebral artery occlusion,MCAO)模型,将造模成功大鼠随机分为模型组(n=36)、脑舒明方低剂量组(n=36)、中剂量组(n=36)及高剂量组(n=36),另设正常组(n=12)及假手术组(n=12)。每组分别于造模后3,7,14 d处死并检测相关指标,参照Ayelet Levy 14分评分法进行神经功能损伤评分,采用免疫组织化学检测核转录因子(nuclear factor kappa-B,NF-κB)/p50,NF-κB/p65,肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和IL-1β的蛋白质表达;采用实时荧光定量PCR检测NF-κB,TNF-α和IL-1β的mRNA表达。结果:与假手术组比较,各个时间点的MCAO模型组及脑舒明方各剂量组大鼠炎性指标显著增强(均P<0.05或P<0.01),都有神经功能障碍,但随着时间推移炎症指标及神经功能障碍会逐渐改善。用药治疗3,7,14 d时,分别与模型组比较,脑舒明方各剂量组大鼠的神经功能评分均得到改善,NF-κB/p50,NF-κB/p65,TNF-α和IL-1β的蛋白质表达,以及NF-κB,TNF-α和IL-1β的mRNA表达均降低,差异均有统计学意义(均P<0.05或P<0.01)。结论:脑舒明方可减轻脑缺血大鼠的损伤。

Objective:To explore the effect of Naoshuming decoction on cerebral ischemic rats.Methods:The model of cerebral ischemia in rats was established via middle cerebral artery occlusion(MCAO).The MCAO model rats were randomly divided into a model group(n=36),a Naoshuming decoction at high dose group(n=36),a Naoshuming decoction at middle dose group(n=36)and a Naoshuming decoction at low dose group(n=36).In addition,a normal group(n=12)and a sham operation group(n=12)were included.Rats in each group were killed on the 3rd,7th,and 14th day to detect relevant indicators.The Ayelet Levy 14 method was used to score the neurological function.Immunohistochemical method was used to detect the protein expression of nuclear factor kappa-B(NF-κB)/p50,NF-κB/p65,tumor necrosis factor-α(TNF-α),and IL-1β.The quantitative real-time PCR were used to detect the mRNA expression of NF-κB,TNF-αand IL-1β.Results:Compared with the sham group,at each time point,the inflammation indexes in the model group and different dose of Naoshuming decoction groups were significantly enhanced,and all of them showed neurological dysfunction.But the inflammatory indexes and neurological function scores would were gradually improved with the pass of time.Compared with the model group,the neurological dysfunction,the protein levels of NF-κB/p50,NF-κB/p65,TNF-αand IL-1β,and the mRNA of NF-κB,TNF-αand IL-1βin the high,middle and low dose of Naoshuming decoction groups were reduced at 3,7 and 14 d,with statistical difference(all P<0.05 or P<0.01).Conclusion:Naoshuming decoction can alleviate the cerebral ischemic injury in rats.