[1]杨 谦,曹冰清,韩 征,等.CXC趋化因子配体10在癫痫大鼠和细胞炎症反应中的作用及机制研究[J].陕西医学杂志,2022,51(12):1483-1487.[doi:DOI:10.3969/j.issn.1000-7377.2022.12.004]
　YANG Qian,CAO Bingqing,HAN Zheng,et al.Role and mechanism of CXC chemokine ligand-10 in inflammatory response of epileptic rats and cells[J].,2022,51(12):1483-1487.[doi:DOI:10.3969/j.issn.1000-7377.2022.12.004]
点击复制

CXC趋化因子配体10在癫痫大鼠和细胞炎症反应中的作用及机制研究
分享到：

《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年12期
页码:
1483-1487
栏目:
基础研究
出版日期:
2022-12-05

文章信息/Info

Title:
Role and mechanism of CXC chemokine ligand-10 in inflammatory response of epileptic rats and cells
作者:
杨 谦; 曹冰清; 韩 征; 薛延莉; 康 涛
(陕西省人民医院神经内科,陕西 西安 710068)
Author(s):
YANG Qian; CAO Bingqing; HAN Zheng; XUE Yanli; KANG Tao
(Department of Neurology,Shaanxi Provincial People's Hospital,Xi'an 710068,China)
关键词:
癫痫;  CXC趋化因子配体10;  Toll样受体4;  核因子-κB;  炎症因子;  大鼠模型
Keywords:
Epilepsy;  CXC chemokine ligand-10;  Toll-like receptor 4;  Nuclear factor-κB;  Inflammatory factors;  Rat model
分类号:
R 742.1
DOI:
DOI:10.3969/j.issn.1000-7377.2022.12.004
文献标志码:
A
摘要:
目的:探究CXC趋化因子配体10(CXCL10)在癫痫大鼠和细胞炎症反应中的作用及机制。方法:构建大鼠癫痫模型,采用实时荧光定量PCR(qRT-PCR)检测对照组和模型组大鼠中CXCL10表达。无镁细胞液处理SH-SY5Y细胞建立癫痫细胞模型(SH-SY5Y组),采用qRT-PCR检测空白组和SH-SY5Y组中CXCL10表达。用pcDNA3.1-CXCL10或si-CXCL10转染SH-SY5Y细胞,采用酶联免疫吸附(ELISA)法检测pcDNA3.1组、pcDNA3.1-CXCL10组、si-RNA组和si-CXCL10组中炎症细胞因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和IL-8]水平。SH-SY5Y细胞通过转染si-CXCL10或经Toll样受体4(TLR4)抑制剂(TAK-242)和激动剂脂多糖(LPS)处理,采用蛋白免疫印迹(Western blot)检测si-RNA组、si-CXCL10组、si-CXCL10+TAK-242组和si-CXCL10+LPS组中TLR4、NF-κB p-p65/p65蛋白表达水平。结果:模型组大鼠CXCL10 mRNA表达水平高于对照组(P<0.05)。SH-5Y5Y组细胞CXCL10 mRNA水平高于空白组(P<0.05)。过表达CXCL10升高SH-SY5Y细胞CXCL10 mRNA表达水平,而抑制CXCL10降低了SH-SY5Y细胞CXCL10 mRNA表达水平(均P<0.05)。此外,过表达CXCL10使SH-SY5Y细胞培养液中TNF-α、IL-6和IL-8水平显著上升,si-CXCL10使SH-SY5Y细胞 TNF-α、IL-6和IL-8水平显著下降(均P<0.05)。过表达CXCL10促进了TLR4和NF-κB p-p65/p65蛋白表达,而敲低CXCL10可抑制TLR4和NF-κB p-p65/p65蛋白水平(均P<0.05)。与si-CXCL10组比较,经LPS处理的癫痫模型细胞TNF-α、IL-6、IL-8水平上升,而经TAK-242处理的癫痫模型细胞TNF-α、IL-6、IL-8水平显著下降(均P <0.05)。结论:CXCL10在癫痫大鼠和细胞中相对表达量均升高,沉默CXCL10可通过TLR4/NF-κB信号通路抑制炎性细胞因子表达,在癫痫炎症反应过程中发挥重要作用。
Abstract:
Objective:To explore the role and mechanism of CXC chemokine ligand-10(CXCL10)in inflammatory response of epileptic rats and cells.Methods:The rat epilepsy model was established,and the expression of CXCL10 in the control group and model group was detected by qRT-PCR.The cells were treated with magnesium to establish the epilepsy cell model(SH-SY5Y group),and the expression of CXCL10 in the blank group and SH-SY5Y group was detected by qRT-PCR.SH-SY5Y cells were transfected with pcDNA3.1-CXCL10 or si-CXCL10 to detect inflammatory cytokines(including TNF-α,IL-6 and IL-8)in pcDNA3.1 group,pcDNA3.1-CXCL10 group,si-RNA group and si-CXCL10 group by ELISA.SH-SY5Y cells were transfected with si-CXCL10 or treated with Toll-like receptor 4(TLR4)inhibitor(TAK-242)and agonist(LPS),and Western blot was used to detect the protein expression of TLR4 and NF-κB p-p65/p65 in si-RNA group,si-CXCL10 group,si-CXCL10+TAK-242 group and si-CXCL10+LPS group.Results:The expression level of CXCL10 mRNA in model group was higher than that in control group(P<0.05).The level of CXCL10 mRNA in SH-5Y5Y group was higher than that in blank group(P<0.05).Overexpression of CXCL10 increased CXCL10 mRNA expression in SH-SY5Y cells,whereas inhibition of CXCL10 decreased CXCL10 mRNA expression in SH-SY5Y cells(all P<0.05).In addition,overexpression of CXCL10 significantly increased the levels of TNF-α,IL-6,and IL-8 in SH-SY5Y cell cultures,and si-CXCL10 decreased the levels of TNF-α,IL-6,and IL-8 in SH-SY5Y cells(all P<0.05).Overexpression of CXCL10 promoted TLR4 and NF-κB p-p65/p65 protein expression,while knockdown of CXCL10 inhibited TLR4 and NF-κB p-p65/p65 protein levels(all P<0.05).Compared with the si-CXCL10 group,the levels of TNF-α,IL-6 and IL-8 in the epileptic model cells treated with LPS were increased,while the levels of TNF-α,IL-6 and IL-8 in the epileptic model cells treated with TAK-242 were significantly decreased(all P<0.05).Conclusion:The relative expression of CXCL10 is increased in both epileptic rats and cells.Silencing CXCL10 can inhibit the expression of inflammatory cytokines through TLR4/NF-κB signaling pathway,which plays an important role in the inflammatory response of epilepsy.

参考文献/References:

[1] Xiang J,Wen F,Zhang L,et al.FOXD3 inhibits SCN2A gene transcription in intractable epilepsy cell models[J].Experimental Neurology,2018,302:14-21.
[2] Wang L,Song LF,Chen XY,et al.MiR-181b inhibits P38/JNK signaling pathway to attenuate autophagy and apoptosis in juvenile rats with kainic acid-induced epilepsy via targeting TLR4[J].CNS Neuroscience & Therapeutics,2019,25(1):112-122.
[3] Fu J,Tao T,Li Z,et al.The roles of ER stress in epilepsy:Molecular mechanisms and therapeutic implications[J].Biomedicine & Pharmacotherapy,2020,131:110658.
[4] Pernhorst K,Herms S,Hoffmann P,et al.TLR4,ATF-3 and IL-8 inflammation mediator expression correlates with seizure frequency in human epileptic brain tissue[J].Seizure,2013,22(8):675-678.
[5] Matin N,Tabatabaie O,Falsaperla R,et al.Epilepsy and innate immune system:A possible immunogenic predisposition and related therapeutic implications[J].Hum Vaccin Immunother,2015,11(8):2021-2029.
[6] Zhang B,Zou J,Rensing NR,et al.Inflammatory mechanisms contribute to the neurological manifestations of tuberous sclerosis complex[J].Neurobiology of Disease,2015,80:70-79.
[7] Wang Y,Li Z.RNA-seq analysis of blood of valproic acid-responsive and non-responsive pediatric patients with epilepsy[J].Experimental and Therapeutic Medicine,2019,18(1):373-383.
[8] Paudel YN,Angelopoulou E,Akyuz E,et al.Role of innate immune receptor TLR4 and its endogenous ligands in epileptogenesis[J].Pharmacological Research,2020,160:105172.
[9] Kan M,Song L,Zhang X,et al.Circulating high mobility group box-1 and toll-like receptor 4 expressions increase the risk and severity of epilepsy[J].Brazilian Journal of Medical and Biological Research,2019,52(7):e7374.
[10] Tülay K,Beril D,Serap-Özer Y,et al.HMGB-1,TLR4,IL-1R1,TNF-α,and IL-1β:Novel epilepsy markers?[J].Epileptic Disorders,2020,22(2):183-193.
[11] Qu Z,Jia L,Xie T,et al.Epigallocatechin-3-gallate protects against lithium-pilocarpine-induced epilepsy by inhibiting the toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway[J].Med Sci Moni,2019,25:1749-1758.
[12] 张亚琼,姬冬林.奥扎格雷钠对癫痫小鼠应激、神经损伤程度和神经功能重建相关因子及凝血指标的影响[J].陕西医学杂志,2022,51(7):787-790.
[13] 王 倩,曾 培,袁莉莉,等.柴胡加龙骨牡蛎汤调控NF-κB信号通路对癫痫大鼠神经炎症及海马神经元损伤的影响[J].陕西中医,2021,42(10):1327-1332,1362.
[14] 刘源源.左乙拉西坦治疗成人癫痫疗效及对患者血清脑源性神经营养因子和miR-134-5p的影响[J].陕西医学杂志,2021,50(3):349-352.
[15] Manford M.Recent advances in epilepsy[J].J Neurol,2017,264(8):1811-1824.
[16] Shi Y,Zhang L,Teng J,et al.HMGB1 mediates microglia activation via the TLR4/NF-κB pathway in coriaria lactone induced epilepsy[J].Molecular Medicine Reports,2018,17(4):5125-5131.
[17] Sorensen EW,Lian J,Ozga AJ,et al.CXCL10 stabilizes T cell-brain endothelial cell adhesion leading to the induction of cerebral malaria[J].JCI Insight,2018,3(8):e98911.
[18] Bradburn S,Mcphee J,Bagley L,et al.Dysregulation of C-X-C motif ligand 10 during aging and association with cognitive performance[J].Neurobiology of Aging,2018,63:54-64.
[19] Wilson NO,Solomon W,Anderson L,et al.Pharmacologic inhibition of CXCL10 in combination with anti-malarial therapy eliminates mortality associated with murine model ofcerebral malaria[J].PLoS One,2013,8(4):e60898.
[20] Zhang Y,Thai K,Kepecs DM,et al.Reversing CXCL10 deficiency ameliorates kidney disease in diabetic mice[J].Am J Pathol,2018,188(12):2763-2773.
[21] Berger TC,Vigeland MD,Hjorthaug HS,et al.Differential glial activation in early epileptogenesis-insights from cell-specific analysis of DNA methylation and gene expression in the contralateral hippocampus[J].Frontiers in Neurology,2020,11:573575.
[22] Yu L,Yang J,Yu W,et al.Rhein attenuates PTZ induced epilepsy and exerts neuroprotective activity via inhibition of the TLR4-NFκB signaling pathway[J].Neuroscience Letters,2021,758:136002.
[23] Liu L,Xu Y,Dai H,et al.Dynorphin activation of kappa opioid receptor promotes microglial polarization toward M2 phenotype via TLR4/NF-κB pathway[J].Cell & Bioscience,2020,10:42-52.
[24] Kleen JK,Holmes GL.Taming TLR4 may ease seizures[J].Nature Medicine,2010,16(4):369-370.
[25] Iori V,Maroso M,Rizzi M,et al.Receptor for advanced glycation endproducts is upregulated in temporal lobe epilepsy and contributes to experimental seizures[J].Neurobiology of Disease,2013,58:102-114.
[26] Dey A,Kang X,Qiu J,et al.Anti-inflammatory small molecules to treat seizures and epilepsy:From bench to bedside[J].Trends in Pharmacological Sciences,2016,37(6):463-484.

相似文献/References:

[1]史剑飞,卢北玲,宁远征△.儿童癫痫患者血清胱抑素C表达水平及检测意义*[J].陕西医学杂志,2019,(10):1393.
[2]刘源源.左乙拉西坦治疗成人癫痫疗效及对患者血清脑源性神经营养因子和miR-134-5p的影响[J].陕西医学杂志,2021,50(3):349.[doi:DOI:10.3969/j.issn.1000-7377.2021.03.024]
　LIU Yuanyuan.Efficacy of levetiracetam in treatment of adult epilepsy and its effect on serum BDNF and miR-134-5p[J].,2021,50(12):349.[doi:DOI:10.3969/j.issn.1000-7377.2021.03.024]
[3]张亚琼,姬冬林.奥扎格雷钠对癫痫小鼠应激、神经损伤程度和神经功能重建相关因子及凝血指标的影响[J].陕西医学杂志,2022,51(7):787.[doi:DOI:10.3969/j.issn.1000-7377.2022.07.004]
　ZHANG Yaqiong,JI Donglin.Effects of sodium ozagrel on stress,nerve injury degree,neural function reconstruction related factors and coagulation indexes in epilepsy mice[J].,2022,51(12):787.[doi:DOI:10.3969/j.issn.1000-7377.2022.07.004]
[4]曹志伟,张润春,王玉珍,等.癫痫患儿血清胱抑素C、脂蛋白相关磷脂酶A2水平与拉考沙胺添加治疗反应性关系及其预测价值[J].陕西医学杂志,2024,(3):372.[doi:DOI:10.3969/j.issn.1000-7377.2024.03.018]
[5]朱芳丽,马厉英,马晓莹,等.急性胰腺炎患者血清CXC趋化因子配体10和CC类趋化因子22水平与疾病严重程度关系及临床诊断价值研究[J].陕西医学杂志,2024,(6):788.[doi:DOI:10.3969/j.issn.1000-7377.2024.06.014]
　ZHU Fangli,MA Liying,MA Xiaoying,et al.Relationship between serum CXCL10,CCL22 levels and disease severity in patients with acute pancreatitis and their clinical diagnostic value[J].,2024,(12):788.[doi:DOI:10.3969/j.issn.1000-7377.2024.06.014]
[6]刘维伟,王翠敏,刘平安,等.糖尿病足感染患者血清CXC趋化因子配体10和可溶性髓系细胞触发受体-1水平与病情、治疗效果的关系[J].陕西医学杂志,2025,54(3):349.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.012]
　LIU Weiwei,WANG Cuimin,LIU Ping'an,et al.Relationship between serum CXCL10,sTREM-1 levels and disease condition,therapeutic efficacy in patients with diabetic foot infection[J].,2025,54(12):349.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.012]
[7]于 乐,王 沛,姜梦真.癫痫患儿血清基质金属蛋白酶-9、谷氨酸脱羧酶抗体、高迁移率族蛋白B1水平与左乙拉西坦治疗效果相关性及联合检测对治疗效果的预测价值[J].陕西医学杂志,2025,54(3):374.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.017]
　YU Le,WANG Pei,JIANG Mengzhen.Correlation of serum MMP-9,GADA,HMGB1 levels with therapeutic effect of levetiracetam in children with epilepsy,and predictive value of combined detection for therapeutic effect[J].,2025,54(12):374.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.017]

备注/Memo

备注/Memo:
基金项目:陕西省自然科学基础研究计划项目(2022JM-590)
更新日期/Last Update: 2022-12-05