摘要
目的基于磷脂酰肌醇3-激酶(PI3K)/丝苏氨酸蛋白激酶(AKT)通路探究槲皮素对脂多糖(LPS)诱导的人小胶质细胞(HMC3)增殖、迁移和M1型极化的影响。方法将HMC3细胞分为对照组(正常培养)、模型组(炎症模型)、槲皮素组(造模后加入10.0μmol·L^(-1)槲皮素)、胰岛素样生长因子-Ⅰ(IGF-I)组(造模后再加入50.0 mg·mL^(-1)IGF-Ⅰ)、激动药组(造模后再加入10.0μmol·L^(-1)槲皮素+50.0 mg·mL^(-1)IGF-Ⅰ)和抑制药组(造模后再加入10.0μmol·L^(-1)槲皮素+10.0μmol·L^(-1)LY294002)。用细胞计数试剂盒-8(CCK-8)检测细胞活力;用酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-10、肿瘤坏死因子(TNF-α)水平;用蛋白质印迹法检测精氨酸酶1(Arg1)和PI3K/AKT通路相关蛋白水平。结果对照组、模型组、槲皮素组、IGF-I组、激动药组和抑制药组的增殖率分别为(42.46±3.37)%、(15.21±1.37)%、(26.86±0.21)%、(25.31±1.91)%、(35.63±1.55)%和(15.79±1.18)%,迁移细胞数分别为68.67±3.51、155.33±6.51、109.33±2.08、95.33±4.62、75.67±6.66和147.33±4.73,IL^(-1)0水平分别为(44.26±1.30)、(17.84±0.82)、(26.26±1.00)、(28.74±0.87)、(37.00±1.24)和(21.56±0.94)ng·L^(-1),TNF-α水平分别为(2.81±0.57)、(18.91±1.01)、(12.04±0.57)、(11.52±0.77)、(5.81±0.50)和(16.56±1.29)pg·mL^(-1),Arg1蛋白表达水平分别为0.95±0.01、0.68±0.01、0.95±0.01、0.90±0.01、1.18±0.03和0.79±0.01,p-PI3K蛋白表达水平分别为0.63±0.02、0.13±0.01、0.37±0.02、0.33±0.02、0.47±0.01和0.19±0.02,p-AKT蛋白表达水平分别为0.69±0.02、0.16±0.01、0.41±0.01、0.42±0.01、0.52±0.01和0.21±0.01。模型组的上述指标与对照组比较,差异均有统计学意义(均P<0.05);槲皮素组的上述指标和IGF-I组与模型组比较,差异均有统计学意义(均P<0.05);激动药组的上述指标和抑制药组与槲皮素组比较,差异均有统计学意义(均P<0.05)。结论槲皮素促进PI3K/AKT信号通路活化提升LPS诱导的HM
Objective To investigate the effects of quercetin on lipopolysaccharide(LPS)-induced proliferation,migration and M1-type polarization of human microglia(HMC3)based on the phosphatidylinositol 3-kinase(PI3K)/serthreonine protein kinase(AKT)pathway.Methods HMC3 cells were divided into control group(normal culture),model group(inflammatory model),quercetin group(adding 10.0μmol·L^(-1)quercetin after model),insulin-like growth the factor-I(IGF-I)group(adding 50.0 mg·m L^(-1)IGF-Ⅰafter model),agonist group(adding 10.0 after model)μmol·L^(-1)quercetin+50.0 mg·m L^(-1)IGF-Ⅰ)and inhibitor group(10.0μmol·L^(-1)quercetin+10.0μmol·L^(-1)LY294002 was added after molding).Cell viability was measured by living cell counting kit(CCK)-8;the levels of interleukin-10(IL-10)and tumor necrosis factor(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA);the levels of arginase 1(Arg1)and PI3K/AKT pathway associated proteins were detected by Western blot.Results The proliferation rates of control group,model group,quercetin group,IGF-I group,agonist group and inhibitor group were(42.46±3.37)%,(15.21±1.37)%,(26.86±0.21)%,(25.31±1.91)%,(35.63±1.55)%and(15.79±1.18)%,respectively;the number of migrating cells were68.67±3.51,155.33±6.51,109.33±2.08,95.33±4.62,75.67±6.66 and 147.33±4.73,respectively;the expression levels of IL-10 were(44.26±1.30),(17.84±0.82),(26.26±1.00),(28.74±0.87),(37.00±1.24)and(21.56±0.94)ng·L^(-1),respectively;the expression levels of TNF-αwere(2.81±0.57),(18.91±1.01),(12.04±0.57),(11.52±0.77),(5.81±0.50)and(16.56±1.29)pg·m L^(-1),respectively;Arg1 protein levels were 0.95±0.01,0.68±0.01,0.95±0.01,0.90±0.01,1.18±0.03 and 0.79±0.01,respectively;p-PI3K protein levels were 0.63±0.02,0.13±0.01,0.37±0.02,0.33±0.02,0.47±0.01 and 0.19±0.02,respectively;p-AKT protein levels were 0.69±0.02,0.16±0.01,0.41±0.01,0.42±0.01,0.52±0.01 and0.21±0.01,respectively.There were statistical significance between model group and control group(all P<0.05);quercetin group an
作者
张颖
王磊
永莲
樛秀俊
杨帆
ZHANG Ying;WANG Lei;YONG Lian;JIU Xiu-jun;YANG Fan(Department of Encephalopathy,Hulan Buir Zhong Meng Hospital,Hulunbeier 021000,Inner Mongolia Autonomous Region,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第22期3276-3280,共5页
The Chinese Journal of Clinical Pharmacology
基金
内蒙古自治区科技厅基金资助项目(2022YFSH0003)
呼伦贝尔市科技局基金资助项目(SF2021007)。
