论文部分内容阅读
目的探讨高糖对肺腺癌A549细胞血红素加氧酶-1(HO-1)表达的影响及机制。方法应用Western blot技术和逆转录PCR技术检测高糖对人肺腺癌上皮细胞系A549细胞HO-1的表达。应用酶联免疫吸附试验检测HO-1酶活性和氧化应激产物。结果用25 mmol/L高糖处理肺A549细胞0、24、48、72 h,以及用5、10、25、40 mmol/L葡萄糖处理A549细胞48 h,在蛋白水平和RNA水平,高糖诱导肺A549细胞HO-1表达呈浓度和时间依赖性。高糖诱导A549细胞活性氧(ROS)和转化生长因子β_1(TGF-β_1)产生增加,并介导HO-1表达增加。伴随HO-1表达增加,HO-1酶活性也相应增加。抗氧化剂N-乙酰半胱氨酸(NAC)和PI3K/Akt抑制剂可抑制高糖诱导的肺上皮细胞HO-1表达。结论高糖促进肺上皮细胞ROS和TGF-β_1产生,介导HO-1表达增加,并伴随HO-1酶活性增加。
Objective To investigate the effect and mechanism of high glucose on the expression of heme oxygenase-1 (HO-1) in lung adenocarcinoma A549 cells. Methods The expression of HO-1 in human lung adenocarcinoma cell line A549 was detected by Western blot and reverse transcription PCR. Enzyme-linked immunosorbent assay was used to detect HO-1 enzyme activity and oxidative stress products. Results A549 cells were treated with 25 mmol / L high glucose for 0, 24, 48, and 72 h, and A549 cells were treated with 5, 10, 25 and 40 mmol / L glucose for 48 h at high protein and RNA levels The lung A549 cell HO-1 expression in a concentration and time-dependent manner. High glucose induced the production of reactive oxygen species (ROS) and transforming growth factor-β 1 (TGF-β 1) in A549 cells and increased the expression of HO-1. With the increase of HO-1 expression, HO-1 enzyme activity also increased accordingly. Antioxidants N-acetylcysteine (NAC) and PI3K / Akt inhibitors can inhibit high glucose-induced HO-1 expression in lung epithelial cells. Conclusion High glucose can promote the production of ROS and TGF-β 1 in lung epithelial cells, and increase the expression of HO-1 with the increase of HO-1 enzyme activity.