毛蕊异黄酮介导GP130/JAK/STAT通路对脊髓星形胶质细胞氧化损伤的影响  

Effect of mullein isoflavones mediated GP130/JAK/STAT pathway on oxidative injury of spinal astrocytes

作　　者：宋颖军[1] 李旭[1] 刘小舟 张国福[1] SONG Ying-Jun;LI Xu;LIU Xiao-Zhou(Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,Jiangxi,China)

机构地区：[1]江西中医药大学附属医院,江西南昌330006

出　　处：《中国老年学杂志》2023年第9期2145-2150,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81860856)。

摘　　要：目的探究毛蕊异黄酮介导糖蛋白(GP)130/Janus酪氨酸蛋白激酶(JAK)/信号转导及转录激活因子(STAT)通路对脊髓星形胶质细胞损伤的影响。方法原代分离大鼠脊髓星形胶质细胞,并利用免疫荧光检测胶原纤维酸性蛋白(GFAP)。分别加入50、100、150μmol/L毛蕊异黄酮预处理脊髓星形胶质细胞12 h,然后加入H_(2)O_(2)处理24 h造氧化损伤模型,CCK8检测各组细胞增殖情况,选择合适的毛蕊异黄酮处理浓度。实验分组:空白对照(Control)组、模型(H_(2)O_(2))组、模型+毛蕊异黄酮预处理(H_(2)O_(2)+Calycosin)组、模型+毛蕊异黄酮预处理+抑制剂Ly294002(H_(2)O_(2)+Calycosin+Ly294002)组、模型+毛蕊异黄酮处理+抑制剂Stattic(H_(2)O_(2)+Calycosin+Stattic)组。CCK8检测各组细胞增殖情况,流式检测各组细胞凋亡和周期情况,免疫荧光检测各组细胞样本中Brdu水平;Western印迹检测各组细胞中p-JAK2、p-STAT3、p-蛋白激酶B(AKT)、GP130、白细胞介素(IL)-6蛋白表达水平。结果H_(2)O_(2)处理能够抑制脊髓星形胶质细胞的增殖并诱导其凋亡,氧化损伤模型细胞组中p-JAK2、p-STAT3、p-AKT、GP130、IL-6的蛋白表达水平显著增加,而毛蕊异黄酮预处理后能够减轻H_(2)O_(2)造成的氧化损伤,促进增殖和抑制凋亡,并显著抑制p-JAK2、p-STAT3、p-AKT、GP130、IL-6蛋白表达水平(P<0.05)。结论蕊异黄酮能够促进氧化损伤的星形胶质细胞增殖并抑制其凋亡,并能通过抑制磷脂酰肌醇-3激酶(PI3K)/AKT通路磷酸化、JAK2/STAT3通路磷酸化起作用。Objective To explore the effects of mullein isoflavones mediated glycoprotein(GP)130/Janus tyrosine protein kinase(JAK)/signal transduction and transcriptional activators(STAT)pathway on spinal cord astrocytes injury.Methods Primary isolation of rat spinal astrocytes and glial fibrillary acidic protein(GFAP)was detected by immunofluorescence.The spinal astrocytes were pretreated with 50,100,150μmol/L of mullein isoflavones for 12 h,and then treated with H_(2)O_(2) for 24 h to induce oxidative damage model.The cell proliferation of each group was detected by CCK8 and the appropriate concentration of mullein isoflavones was selected.The experimental groups included blank control(Control)group,model(H_(2)O_(2))group,model+mullein isoflavones pretreatment(H_(2)O_(2)+Calycosin)group,model+mullein isoflavones pretreatment+inhibitor Ly294002(H_(2)O_(2)+Calycosin+Ly294002)group and model+mullein isoflavones treatment+inhibitor Stattic(H_(2)O_(2)+Calycosin+Stattic)group.Cell proliferation in each group was detected by CCK8,cell apoptosis and cell cycle were detected by flow cytometry,Brdu level in cell sample was detected by immunofluorescence.The protein expression levels of p-JAK2,p-STAT3,p-protein kinase B(AKT),GP130 and interleukin(IL)-6 in each group were detected by Western blot.Results H_(2)O_(2) treatment could inhibit the proliferation and induce the apoptosis of spinal astrocytes,and the protein expression levels of p-JAK2,p-STAT3,p-AKT,GP130 and IL-6 in the oxidative injury model group were significantly increased,while the preconditioning of mullein isoflavones could reduce the oxidative damage caused by H_(2)O_(2),promote the proliferation and inhibit the apoptosis,and the protein expression levels of p-JAK2,p-STAT3,p-AKT,GP130 and IL-6 were significantly inhibited(P<0.05).Conclusions Mullein isoflavones could promote the proliferation and inhibit the apoptosis of oxidative injured astrocytes,and could inhibit the phosphorylation of phosphatidylinositol-3 kinase(PI3K)/AKT pathway and JAK2/STAT3 pathway.

关 键 词：脊髓星形胶质细胞 毛蕊异黄酮 氧化损伤 糖蛋白(GP130)/Janus酪氨酸蛋白激酶(JAK)/信号转导及转录激活因子(STAT)通路 

分 类 号：R446[医药卫生—诊断学]