布地格福调节c-Jun表达抑制肺泡巨噬细胞NR8383氧化应激损伤的机制研究  

Study on the mechanism of regulating c-Jun expression by budesonide to inhibit oxidative stress damage in alveolar macrophages NR8383

作　　者：张孝侠[1] 吴友涛 ZHANG Xiao-xia;WU You-tao(Laboratory of Medical Laboratory,Jining Medical UniversityJ,ining 272067,Shandong Province,China;Department of Emergency,Jining No.2 People'sHospital,Jining 272049,Shandong Province,China)

机构地区：[1]济宁医学院医学检验实验室,山东济宁272067 [2]济宁市第二人民医院急诊科,山东济宁272049

出　　处：《中国临床药理学杂志》2024年第4期534-538,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的 研究布地格福对SD大鼠NR8383氧化应激损伤保护机制。方法 将大鼠肺泡巨噬细胞NR8383分为5组:空白组给予无血清K12培养基,不作任何处理;脂多糖组(LPS组)给予2.29μg·mL^(-1) LPS标准品溶液;布地格福组、c-Jun抑制药组(AS601245)、布地格福+c-Jun抑制药组(Budesonide+AS601245)在LPS组基础上分别给予布地格福28.0μL、c-Jun抑制药AS601245 2.50μg、布地格福28.0μL+c-Jun抑制药AS601245 2.50μg。5组细胞均置于无血清K12培养基恒温培养24 h。以细胞计数试剂盒-8(CCK-8)法检测24 h细胞凋亡率,以实时荧光定量聚合酶链反应(q-PCR)检测c-Jun mRNA表达,以酶联免疫吸附实验(ELISA)法检测氧化应激损伤因子活性氧(ROS)、8-羟基脱氧鸟苷(8-OHdG)、谷胱甘肽过氧化物酶(GSH-Px)、硫氧还蛋白(TRX-1)表达,以蛋白质印迹(Western Blot)法检测各组c-Jun信号通路蛋白的表达。结果 与LPS组(29.88±5.98)%相比,布地格福组、c-Jun抑制药组、布地格福+c-Jun抑制药组、空白组24 h细胞凋亡率显著下降[(20.15±6.66)%、(15.39±3.54)%、(12.11±2.55)%和(8.52±1.27)%],在统计学上差异均有统计学意义(均P<0.05)。布地格福组、c-Jun抑制药组、布地格福+c-Jun抑制药组、LPS组、空白组ROS浓度分别为(3.16±0.19)、(4.15±0.33)、(2.21±0.21)、(6.52±0.36)和(1.06±0.23) U·g^(-1);8-OHdG分别为(10.55±1.23)、(11.14±1.06)、(9.55±1.00)、(15.66±1.99)和(8.27±1.13)ng·mL^(-1);GSH-Px分别为(188.52±12.33)、(200.52±27.97)、(144.52±20.55)、(335.14±30.10)和(126.55±12.52)U·mL^(-1);TRX-1分别为(40.11±6.66)、(50.55±10.07)、(60.25±10.55)、(115.36±20.03)和(16.55±2.33)ng·mL^(-1);c-Jun mRNA相对表达水平分别为0.56±0.03、0.44±0.11、0.25±0.04、0.89±0.12和0.08±0.01;c-Jun/GAPDH蛋白相对表达水平为3.15±0.22、2.36±0.14、1.55±0.13、4.02±0.22和0.88±0.12;与LPS组相比,布地格福组、c-Jun抑制药组、布地格福+c-Jun抑制药组指标差异均显著下降,在统计学上�Objective To investigate the protective mechanism of oxidative stress injury of SD rats NR8383 by budesonide.Methods Rat alveolar macrophages NR8383 were divided into 5 groups:blank group was given serum-free K12 medium without any treatment;lipopolysaccharide(LPS) group was given 2.29 μg · mL^(-1) LPS standard solution;budesonide group(budesonide),c-Jun inhibitor group(AS601245) and budesonide+c-Jun inhibitor group(budesonide+AS601245) were given budesonide 28.0 μL,c-Jun inhibitor AS601245 2.50 μg,and budesonide 28.0 μL+c-Jun inhibitor AS601245 2.50 μg based on the LPS group,respectively.Cells in 5 groups were incubated in serum-free K12 medium at constant temperature for 24 h.The apoptosis rate at 24 h was examined by cell counting kit-8(CCK-8)assay;c-Jun mRNA expression was detected by real-time quantitative polymerase chain reaction(q-PCR);oxidative stress damage factor reactive oxygen species(ROS),8-hydroxy-2-deoxyguanosine(8-OHdG),glutathione peroxidase(GSH-Px),thioredoxin reductase-1(TRX-1) expression were detected by enzymelinked immunosorbent assay(ELISA);c-Jun signaling pathway protein expression in each group by Western blot.Results Compared with LPS group(29.88±5.98) %,24 h apoptosis rate was significantly decreased in budesonide group,c-Jun inhibitor group,budesonide+c-Jun inhibitor group and blank group [(20.15±6.66) %,(15.39±3.54) %,(12.11±2.55) % and(8.52±1.27) %,respectively],the differences were statistically significant(all P <0.05).The ROS in budesonide group,c-Jun inhibitor group,budesonide+c-Jun inhibitor group,LPS group and blank group were(3.16±0.19),(4.15±0.33),(2.21±0.21),(6.52±0.36) and(1.06±0.23) U·g^(-1);8-OHdG were(10.55±1.23),(11.14±1.06),(9.55±1.00),(15.66±1.99) and(8.27±1.13) ng·mL^(-1);GSH-PX were(188.52±12.33),(200.52±27.97),(144.52±20.55),(335.14±30.10)and(126.55±12.52) U·mL^(-1);TRX-1 were(40.11±6.66),(50.55±10.07),(60.25±10.55),(115.36±20.03) and(16.55±2.33) ng · mL^(-1);the relative c-Jun mRNA expressions were 0.56±0.03,0.44±0.11,0.25±0.

关 键 词：布地格福 肺泡巨噬细胞 慢性阻塞性肺疾病 C-JUN蛋白 氧化应激 细胞凋亡 

分 类 号：R97[医药卫生—药品]