摘要
目的探讨姜黄素对高氧诱导肺泡Ⅱ型上皮细胞(AEC2)损伤的保护作用,以及β-catenin/p300相互作用与姜黄素保护AEC2的关系。方法采用分离培养的AEC2建立高氧诱导的细胞损伤模型。培养AEC2细胞,随机分为对照组、高氧组、姜黄素组(分为高、中、低剂量3个亚组)、IQ-1(β-catenin/p300相互作用抑制剂)组、p300 siRNA组、siRNA阴性对照组。药物处理24 h后,CCK-8 法和细胞计数板检测AEC2细胞增殖活力和细胞总数,定量PCR检测AEC2、肺泡Ⅰ型上皮细胞(AEC1)的标记基因SP-C和AQP5 mRNA表达量,流式细胞仪检测AQP5、SP-C阳性细胞百分比,Western blot 检测β-catenin、p300蛋白表达量。结果与对照组比较,高氧组AEC2细胞增殖活力、细胞计数、SP-C mRNA表达以及AEC2百分比均明显减少( P <0.01),而AEC1标记基因AQP5 mRNA表达、AEC1百分比、AEC2向AEC1转分化的中间型细胞百分比则明显增加( P <0.01),β-catenin、p300蛋白表达量明显增加( P <0.01)。姜黄素组、IQ-1组和p300 siRNA组AEC2细胞增殖活力、细胞计数、SP-C mRNA表达以及AEC2百分比均明显增加( P <0.01),而AQP5 mRNA表达、AEC1百分比、转分化中间型细胞百分比均减少( P <0.01),β-catenin、p300蛋白表达量明显减小( P <0.01)。结论姜黄素对高氧诱导培养AEC2损伤具有保护作用,其机制与抑制β-catenin/p300相互作用有关。
Objective To investigate the protective effect of curcumin on hyperoxia-induced injury of alveolar type Ⅱ epithelial cells (AEC2) and the relationship between beta-catenin/p300 and the protective effect of curcumin.Methods The models of hyperoxia-induced cellular injury were established in the primary cultured AEC2.AEC2 were randomly divided into 8 groups including the control group, hyperoxia model group, curcumin treatment group which was subdivided into the high-dose, medium-dose and low-dose groups, IQ-1 (β-catenin/p300 interaction inhibitor) group, p300 siRNA group and siRNA negative control group.After 24 hours of the drug treatment, the proliferation activity of AEC2 was detected by CCK-8 assay and the total amount of AEC2 cells was measured by blood cell counting board.The mRNA expression of the marker genes SP-C for AEC2 and AQP5 for alveolar type I epithelial cells (AEC1) was detected by quantitative Polymerase Chain Reaction (qPCR), the percentage of the AQP5 and SP-C positive cells was detected by flow cytometry, and the protein expression of β-catenin and P300 was detected by Western blot.Results When compared with the control group, there was a significant decrease in the AEC2 proliferation, total cell amount, mRNA expression of the AEC2 marker gene SP-C and percentage of AEC2 ( P <0.01) and a significant increase in the mRNA expression of the AEC1 marker gene AQP5, percentage of AEC1 and percentage of the transdifferentiated intermediate cells from AEC2 to AEC1, and protein expression of β-catenin and p300 ( P <0.01).In contrast, the AEC2 proliferation activity, cell count, SP-C mRNA expression and AEC2 percentage increased significantly ( P <0.01) while the AQP5 mRNA expression, AEC1 percentage, percentage of the transdifferentiated intermediate cells, and protein expression of β-catenin and p300 decreased significantly ( P <0.01) in the curcumin treatment group, IQ-1 group and p300 siRNA group.Conclusion Curcumin can protect AEC2 against the hyperoxia-induced injury in vitro, and the mechanism is related to its inhibition of the interaction between β-catenin and p300.
作者
方恩容
杨凯
何杰
邱静
黄娜
Fang Enrong;Yang Kai;He Jie;Qiu Jing;Huang Na(Department of Respiratory Medicine, The First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, China)
出处
《成都医学院学报》
CAS
2019年第3期303-308,共6页
Journal of Chengdu Medical College
