GSTP1通过调节JNK通路抑制癫痫大鼠海马区星形胶质细胞炎性激活  

GSTP1 inhibits inflammasome activation of astrocytes in the hippocampus of epileptic rats through regulation of the JNK pathway

作　　者：赵增霞 刘波 刘淑云 黄正义 ZHAO Zengxia;LIU Bo;LIU Shuyun(Department of Neurology,Shenzhen Longhua District Central Hospital,Shenzhen 518110,China)

机构地区：[1]深圳市龙华区中心医院神经内科,广东深圳518110 [2]深圳市龙华区中心医院老年医学科,广东深圳518110

出　　处：《中风与神经疾病杂志》2023年第2期138-142,共5页Journal of Apoplexy and Nervous Diseases

基　　金：深圳市龙华区医疗卫生机构区级科研项目(No.2022100)。

摘　　要：目的明确谷胱甘肽硫转移酶P1(GSTP1)对星形胶质细胞炎性激活的影响及相关分子机制。方法10周龄雄性SD大鼠采用腹腔注射氯化锂建立癫痫模型(n=8)并收集海马区脑组织。大鼠原代星形胶质细胞给予不同浓度脂多糖(0μg/ml、0.1μg/ml、1μg/ml、10μg/ml、100μg/ml)诱导处理48 h。利用蛋白质印迹检测组织与细胞中GSTP1、JNK、p-JNK蛋白表达水平,以酶联免疫吸附试验检测TNF-α、IL-1β、IL-6浓度,以高效液相色谱测定谷氨酸(Glu)浓度。将GSTP1过表达载体瞬时转染至脂多糖诱导星形胶质细胞,并给予茴香霉素处理,观察星形胶质细胞炎性激活情况。结果癫痫大鼠海马区脑组织中GSTP1蛋白表达水平低于正常大鼠,而p-JNK蛋白表达水平与TNF-α、IL-1β、IL-6、Glu浓度高于正常大鼠(P<0.05)。GSTP1与p-JNK蛋白表达水平呈负相关(P<0.05)。脂多糖诱导星形胶质细胞炎性激活,表现为GSTP1蛋白表达水平呈剂量依耐性降低,而p-JNK蛋白表达水平与TNF-α、IL-1β、IL-6、Glu浓度呈剂量依耐性升高(P<0.05)。过表达GSTP1抑制脂多糖诱导星形胶质细胞炎性激活,而茴香霉素可以部分逆转GSTP1的抑制作用。结论GSTP1对癫痫大鼠海马区星形胶质细胞炎性激活具有抑制作用,其分子机制与抑制JNK通路激活相关。Objective To investigate the effects and related molecular mechanisms of glutathione S-transferase pi 1(GSTP1)on the inflammasome activation of astrocytes.Methods A model of epilepsy was established in 10-week-old male SD rats by intraperitoneal injection of lithium chloride(n=8),and brain tissues were collected from the hippocampus.Rat primary astrocytes were treated with different concentrations of lipopolysaccharide(0μg/ml,0.1μg/ml,1μg/ml,10μg/ml and 100μg/ml)for 48 h.The protein levels of GSTP1,JNK and p-JNK in tissues and cells were measured by Western blotting.The levels of TNF-α,IL-1β,IL-6 were detected by enzyme-linked immunosorbent assay.The levels of glutamate(Glu)were detected by high pressure liquid chromatography.The lipopolysaccharide-induced astrocytes were transiently transfected with GSTP1 overexpression vector and were treated with Anisomycin(JNK activator),and the inflammatory activation of astrocytes was observed.Results The protein levels of GSTP1 were lower in the hippocampal brain tissues of epileptic rats than those in normal rats,while the protein levels of p-JNK and the levels of TNF-α,IL-1β,IL-6 and Glu were higher in epileptic rats than those in normal rats(P<0.05).GSTP1 was negatively correlated with p-JNK protein expression level(P<0.05).Lipopolysaccharide-induced inflammasome activation in astrocytes,as evidenced by a dose-dependent decrease in the protein expression levels of GSTP1 and a dose-dependent increase in the protein levels of p-JNK and the levels of TNF-α,IL-1β,IL-6 and Glu(P<0.05).Overexpression of GSTP1 inhibited lipopolysaccharide-induced inflammasome activation of astrocytes,while Anisomycin partially reversed the inhibitory effect of GSTP1.Conclusion GSTP1 inhibits inflammasome activation of astrocytes in the hippocampus of epileptic rats,and its molecular mechanism is related to JNK pathway inhibition.

关 键 词：谷胱甘肽硫转移酶P1 癫痫 星形胶质细胞 炎性激活 JNK通路 

分 类 号：R742.1[医药卫生—神经病学与精神病学]