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  

Role and mechanism of CXC chemokine ligand-10 in inflammatory response of epileptic rats and cells

作　　者：杨谦[1] 曹冰清[1] 韩征 薛延莉 康涛[1] YANG Qian;CAO Bingqing;HAN Zheng;XUE Yanli;KANG Tao(Department of Neurology,Shaanxi Provincial People’s Hospital,Xi’an 710068,China)

机构地区：[1]陕西省人民医院神经内科,陕西西安710068

出　　处：《陕西医学杂志》2022年第12期1483-1487,共5页Shaanxi Medical Journal

基　　金：陕西省自然科学基础研究计划项目(2022JM-590)。

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。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 oRsi-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 oRtreated with Toll-like receptoR4(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 higheRthan that in control group(P<0.05).The level of CXCL10 mRNA in SH-5Y5Y group was higheRthan 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

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 

分 类 号：R742.1[医药卫生—神经病学与精神病学]