摘要
目的探讨米诺环素(Min)调控CX3C趋化因子配体1(CX3CL1)/趋化因子CX3C受体1(CX3CR1)信号通路对小鼠蛛网膜下腔出血(SAH)后迟发性脑血管痉挛的影响。方法将84只小鼠随机分为对照组、SAH组、Min低剂量组、Min高剂量组、尼莫地平组、CX3CL1中和抗体(CX3CL1抑制剂)组、Min高剂量+CX3CL1中和抗体组(每组n=12)。除对照组外,其他组小鼠均按照血管内穿线法构建SAH小鼠模型。所有小鼠均在建模前30 min和建模后1 d、2 d、3 d进行给药处理(×4次)。用改良Garcia评分法评估神经功能;ELISA法检测脑脊液中IL-1β、TNF-α水平;苏木精-伊红染色检测小鼠基底动脉病理变化;TUNEL染色检测小鼠基底动脉中内皮细胞凋亡;Western blot检测基底动脉中裂解的cleved-caspase-3、CX3CL1、CX3CR1蛋白表达。结果与对照组比较,SAH组小鼠神经功能评分及CX3CL1、CX3CR1蛋白水平降低,IL-1β、TNF-α水平、内皮细胞凋亡率及cleved-caspase-3蛋白水平升高,基底动脉管腔横截面积缩小、管壁厚度增加(均P<0.05)。与SAH组比较,Min低剂量组、Min高剂量组和尼莫地平组小鼠神经功能评分及CX3CL1、CX3CR1蛋白水平升高,IL-1β、TNF-α、cleved-caspase-3蛋白水平及内皮细胞凋亡率降低,基底动脉管腔横截面积变大、管壁厚度变小;CX3CL1中和抗体组对应指标变化趋势与上述相反(均P<0.05)。与Min高剂量组比较,Min高剂量+CX3CL1中和抗体组小鼠神经功能评分及CX3CL1、CX3CR1蛋白水平降低,IL-1β、TNF-α、cleved-caspase-3蛋白表达水平及内皮细胞凋亡率升高,基底动脉管腔横截面积缩小、管壁厚度增加(均P<0.05)。结论Min可能通过激活CX3CL1/CX3CR1信号通路改善小鼠SAH后脑血管痉挛。
Aim To investigate the impact of minocycline(Min)on delayed cerebral vasospasm after subarachnoid hemorrhage(SAH)in mice by regulating CX3C chemokine ligand 1(CX3CL1)/chemokine CX3C receptor 1(CX3CR1)signaling pathway.Methods Eighty-four mice were randomly grouped into a control group,a SAH group,a Min low dose group,a Min high dose group,a nimodipine group,a CX3CL1 neutralizing antibody(CX3CL1 inhibitor)group,a Min high dose+CX3CL1 neutralizing antibody group(12 mice in each group).Except for the control group,SAH mouse models were constructed using intravascular threading method in all other groups.All mice were administered 30 minutes before modeling and 1,2,and 3 days after modeling(×4 times).The improved Garcia scoring method was applied to evaluate neural function.ELISA method was applied to detect the levels of IL-1βand TNF-αin cerebrospinal fluid.Hematoxylin-eosin staining was applied to detect pathological changes in the basal artery of mice.TUNEL staining was applied to detect the apoptosis of endothelial cells in the basal arteries of mice.Western blot was applied to analyze the protein expression of cleved-caspase-3,CX3CL1 and CX3CR1 in basilar artery.Results Compared with the control group,the neurological function score and the protein levels of CX3CL1 and CX3CR1 in the SAH group decreased,the levels of IL-1β,TNF-α,endothelial cell apoptosis rate,and levels of cleved-caspase-3 protein increased,the crosssectional area of the basilar artery lumen decreased,and the wall thickness increased(P<0.05).Compared with the SAH group,the neurological function score and the protein levels of CX3CL1 and CX3CR1 in the Min low dose group,Min high dose group and nimodipine group increased,IL-1β,TNF-α,clevedcaspase-3 protein levels and endothelial cell apoptosis rate were decreased,the cross-sectional area of the basilar artery lumen increased,and the wall thickness decreased,the change trend of corresponding indicators in the CX3CL1 neutralizing antibody group was opposite to the above(P<0.05);Compared with the Min high dose group,the neurological function score and the protein levels of CX3CL1 and CX3CR1 in the Min high dose+CX3CL1 neutralizing antibody decreased,the expression levels of IL-1β,TNF-α,cleved-caspase-3 protein and the apoptosis rate of endothelial cells were increased,the cross-sectional area of the basilar artery lumen decreased,and the wall thickness increased(P<0.05).Conclusion Min may improve cerebral vasospasm after SAH in mice by activating CX3CL1/CX3CR1 signaling pathway.
作者
李阳阳
方建
王晓雪
LI Yang-yang;FANG Jian;WANG Xiaoxue(Department of Neurology for the Elderly,the First Affiliated Hospital of Henan University,Kaifeng 475001,China;Department of Neurology for the Second Ward,the First Affiliated Hospital of Henan University,Kaifeng 475001,China)
出处
《中国临床神经科学》
2024年第2期159-167,共9页
Chinese Journal of Clinical Neurosciences
基金
2021年度河南省医学科技攻关计划项目(编号:LHGJ20210555)。
