摘要
目的分析米诺环素对三叉神经痛(TN)大鼠三叉神经节卫星胶质细胞(SGCs)炎症反应的影响。方法通过激光化学法激发三叉神经损伤,建立TN大鼠模型。按照随机原则,将24只大鼠分为Ⅰ组(正常大鼠,未给予任何用药)、Ⅱ组(构建TN模型)、Ⅲ组(建立TN模型,并给予米诺环素进行灌胃),每组各8只。比较三组大鼠神经颜面部支配区机械疼痛阈值,并比较三组大鼠三叉神经节白细胞介素-1β(IL-1β)和内核转录因子-κB(NF-κB) mRNA及蛋白表达的差异,对NF-κB采取免疫组化染色处理,并用胶质纤维酸性蛋白染色法以观察SGCs的激活情况。通过硝酸甘油建立TN三叉神经节SGCs的炎症模型,经体外培养大鼠三叉神经节SGCs,并将细胞模型分为A组(常规培养)、B组(给予硝酸甘油0. 55 mmol/L)、C组(给予硝酸甘油0. 55 mmol/L+米诺环素15μmol/L)、D组(给予硝酸甘油0. 55mmol/L+米诺环素30μmol/L)。对各组细胞内NF-κB与IL-1β表达水平进行检测,并用Fluo-3/AM探针负载对SGCs中钙离子Ca^(2+)的浓度进行检测。结果相比Ⅰ组,Ⅱ、Ⅲ组大鼠疼痛阈值均明显下降,而Ⅲ组疼痛阈值较Ⅱ组明显升高(P <0. 05)。相比Ⅰ组,Ⅱ、Ⅲ组大鼠三叉神经节IL-1β和NF-κB蛋白及mRNA表达水平均明显升高,Ⅲ组IL-1β和NF-κB蛋白及mRNA的表达水平较Ⅱ组明显下降(P <0. 05)。Ⅰ组NF-κB阳性细胞数量极少;Ⅱ组NF-κB阳性细胞数量较多;Ⅲ组NF-κB阳性细胞数量较Ⅱ组明显减少。Ⅲ组SGCs细胞数量明显减少,Ⅱ组SGCs细胞数量较Ⅰ、Ⅲ组明显增多。相比B组,A、C、D组NF-κB与IL-1β蛋白表达水平明显下降,其mRNA相对表达量明显下降(P <0. 05)。相比A组,B、C组大鼠Ca^(2+)浓度均明显升高,C、D组Ca^(2+)浓度较B组明显下降(P <0. 05)。结论米诺环素可通过阻滞三叉神经节SGCs的激活,下调炎症因子IL-1β与NF-κB的水平以发挥减轻TN症状的作用。
Objective To investigate the effects of minocycline on the inflammatory response of trigeminal ganglion satellite glia cells(SGCs)in rats with trigeminal neuralgia(TN).Methods Trigeminal nerve injury was induced by laser chemical method and TN rat model was established.24 rats were randomized into groupⅠ(normal rats without any intervention),groupⅡ(establishment of TN model),groupⅢ(establishment of TN model and intragastric administration of minocycline)(n=8 in each group).The mechanical pain threshold of facial nerve innervation area in three groups of rats was compared.The expression of interleukin-1β(IL-1β)and nuclear transcription factor-κB(NF-κB)in trigeminal ganglion of three groups of rats was compared.The expression of NF-κB was under immunohistochemical staining and the activation of SGCs was observed by glial fibrillary acidic protein staining.Inflammatory model of TN trigeminal ganglion SGCs was established by nitroglycerin.Rat trigeminal ganglion SGCs were cultured in vitro.Cell models were divided into group A(routine culture),group B(nitroglycerin 0.55 mmol/L),group C(nitroglycerin 0.55 mmol/L+minocycline 15 mol/L),group D(nitroglycerin 0.55 mmol/L+minocycline 30 mol/L).The expression levels of NF-κB and IL-1βin cells of each group were detected,and the concentration of Ca^2+in SGCs was detected by fluo-3/AM probe loading.Results Compared with group I,the pain threshold of groupⅡ(higher than groupⅢ,P P<0.05)and groupⅢdecreased significantly(P<0.05).Compared with groupⅠ,the expression levels of IL-1βand NF-κB protein and RNA in trigeminal ganglion of rats in groupⅡandⅢ(lower than groupⅡ,P<0.05)were significantly higher(P<0.05).The number of NF-κB positive cells was very few in groupⅠ,more in groupⅡ,and much fewer in groupⅢthan in groupⅡ.The number of SGCs in groupⅢsignificantly decreased,but in groupⅡsignificantly increased than in groupⅠandⅢ.Compared with group B,the expression levels of NF-κB and IL-1βprotein in group A,C and D significantly decreased
作者
郭宇鹏
张婵娟
王晓松
王林
黄乙洋
李其壮
陈国强
GUO Yu-peng;ZHANG Chan-juan;WANG Xiao-song(Department of Neurosurgery,Aviation General Hospital,Beijing 100012,China.)
出处
《临床和实验医学杂志》
2019年第9期901-905,共5页
Journal of Clinical and Experimental Medicine
基金
北京市医药卫生计划支持项目基金批准书(编号:20160105679005)
