丹皮酚减轻晚期糖基化终末产物诱导的人Ⅱ型肺泡上皮细胞损伤

Paeonol attenuates the injury of human type Ⅱ alveolar epithelial cells induced by advanced glycation end product

目的探究丹皮酚(Paeonol,Pae)对晚期糖基化终末产物(advanced glycationend end products,AGEs)诱导的人Ⅱ型肺泡上皮细胞(HEPApiC)损伤的影响及机制。方法用Pae预处理HEPApiC 1 h,然后用AGE-BSA刺激48 h。采用MTT法检测细胞活力;采用TUNEL法和Annexin V-FITC/PI染色法检测细胞凋亡;分别采用DCFH-DA测定法和SOD活性检测试剂盒检测细胞内ROS生成和SOD活性;采用Western blot检测细胞中cleaved-caspase3、Bax和细胞质细胞色素C(Cyt C)水平。结果Pae能抑制AGEs诱导的HEPApiC活力降低、凋亡增加,ROS生成增加和SOD活性降低。另外,在HEPApiC中,Pae能抑制AGEs诱导的Cleaved-caspase3、Bax和细胞质Cyt C水平升高。结论Pae通过抑制AGEs诱导的氧化应激和线粒体依赖性凋亡途径来抑制HEPApiC的凋亡。

Objective To investigate the effect of paeonol(Pae) on advanced glycationend end products(AGEs)-induced cell injury in human type Ⅱ alveolar epithelial cells(HEPApiCs) and its mechanism. Methods HEPApi Cs were pretreated with Pae for 1 h, then stimulated with AGE-BSA for 48 h. Cell viability was measured by MTT assay;cell apoptosis was detected by TUNEL staining and Annexin V-FITC/PI staining;intracellular ROS production and SOD activity were detected by DCFH-DA assay and SOD activity detection kit, respectively;the expression levels of cleaved caspase-3, Bax and cytoplasmic cytochrome C(Cyt C) were detected by Western blot. Results Pae inhibited the decrease of cell viability, increase of apoptosis, increase of ROS production and decrease of SOD activity of HEPApiCs induced by AGEs. In addition, Pae inhibited AGEs-induced increase of cleaved caspase3, Bax and cytoplasmic Cyt C levels in HEPApiCs. Conclusion Pae inhibits the apoptosis of HEPApiCs by inhibiting AGEs-induced oxidative stress and mitochondrial dependent apoptosis pathway.