首页|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

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

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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2.29 μg·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28.0 μL、c-Jun抑制药AS601245 2.50 μg、布地格福 28.0 μL+c-Jun 抑制药 AS601245 2.50 μg.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,在统计学上差异均有统计学意义(均P<0.05).结论 布地格福显著增加NR8383细胞的存活率,并显著降低氧化损伤代表因子ROS、GSH-Px、8-OHdG、TRX-1的浓度,其氧化损伤保护机制可能与调控c-Jun蛋白相关.
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 enzyme-linked 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±0.04,0.89±0.12 and 0.08±0.01;c-Jun/GAPDH protein relative expression were 3.15±0.22,2.36±0.14,1.55±0.13,4.02±0.22 and 0.88±0.12.Compared with the LPS group,the differences of indicators in the budesonide group,c-Jun inhibitor group and budesonide+c-Jun inhibitor group possessed statistical significance(all P<0.05).Conclusion Budesonide significantly increased the survival rate of NR8383 cells and significantly decreased the concentrations of oxidative damage representative factors ROS,GSH-Px,8-OHdG and TRX-1,its oxidative damage protection mechanism may be related to the regulation of c-Jun protein.

budesonidealveolar macrophageschronic obstructive pulmonary diseasec-Jun proteinoxidative stresscell apoptosis

张孝侠、吴友涛

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济宁医学院医学检验实验室,山东济宁 272067

济宁市第二人民医院急诊科,山东济宁 272049

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

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(4)
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