电刺激小脑顶核对局灶脑缺血再灌注大鼠脑组织核因子-κB/p65、TNF-α蛋白和基因表达的影响

Effect of electrical stimulation to cerebellar fastigial nucleus on the protein and gene expressions of nuclear factor kappa B/p65 and TNF-α after focal cerebral ischemia reperfusion in rat brain

目的观察电刺激小脑顶核干预对局灶性缺血再灌注大鼠脑组织核因子-κB/p65(NF-κB/p65)、肿瘤坏死因子-α(TNF-α)蛋白和基因表达的影响及其对中枢神经元保护的作用机制。方法 2014年9月—2015年9月在重庆医科大学神经病学实验室进行。Wistar大鼠60只随机分为假手术组(NC组)、缺血再灌注组(I/R组)、电刺激小脑顶核组(FNS组),每组20只,采用线栓法制作脑缺血再灌注动物模型,FNS组予以电刺激小脑顶核干预,造模或干预成功后处死大鼠取脑组织,分别采用免疫组织化学法和RT-PCR检测各组NF-κB/p65、TNF-α的表达。结果与NC组比较,I/R组NF-κB/p65、TNF-α蛋白表达增加(q=0.032、0.020,P<0.05);与I/R组比较,FNS组NF-κB/p65、TNF-α的表达明显降低(q=0.182、0.013,P<0.05)。与NC组比较,L/R组NF-κB/p65mRNA和TNF-αmRNA表达量明显增高(P<0.05);与I/R组比较,FNS组NF-κB/p65 mRNA和TNF-αmRNA表达量明显减少(P均<0.05)。结论电刺激小脑顶核抑制NF-κB/p65的活化和TNF-α的转录,NF-κB/p65的活化抑制和表达减少及所致的TNF-α表达下调可能是其发挥中枢神经源性保护作用的机制之一。

Objective To investigate the mechanism of neuroprotection of cerebellar fastigial nucleus electrical stimulation（FNS） by means of observation of the effect of FNS on the protein and gene expressions of NF-κB/p65 and TNF-a after focal cerebral ischemia reperfusion in rat brain.Methods In the Medical University of Chongqing Laboratory of Neurology,from September 2014 to September 2015,sixty Wistar rats were randomly divided into normal group（NC）,cerebral ischemia reperfusion group（I/R） and cerebral ischemia reperfusion group with a FNS treatment（FNS）,models of focal cerebral ischemia reperfusion injury were established by middle cerebral artery occlusion（MCAO）.Expression of NF-KB/p65 and TNFa was measured by RT-PCR and immunohistochemistry.The absorbance value was measured by imaging analysis.Results Ischemia reperfusion group compared with the normal group,the expression of NF-κB/p65 and TNF-α increased（q =0.032,q =0.020,P〈0.05）,electrical stimulation of fastigial nuclear intervention,NF-κB/p65 and TNF-α expression were significantly reduced,and ischemia reperfusion group were significantly different（q =0.182,q =0.013,P〈0.05）.Compared with the NC group,the expression of NF kappa B/p65 mRNA and TNF-a mRNA was significandy higher in group I/R（P〈0.05）.The expression of NF kappa mRNA and TNF in FNS group was significandy decreased（P〈0.05） compared with I/R group.Conclusion Fastigial nucleus stimulation inhibits NF-κB/p65 activation and TNF-a transcription,NF-κB/p65 activation and expression was inhibited due to the reduction and the expression of TNF-a may be its one of the mechanisms of central nervous protective effects.