胡桃醌抑制Ⅱ型肺泡上皮细胞凋亡减轻体外冲击波导致的细胞损伤

Juglone inhibited alveolar epithelial cells type Ⅱ A549 apoptosis caused by shock waves in vitro

目的探讨胡桃醌对体外冲击波致Ⅱ型肺泡上皮细胞损伤的保护作用及可能机制。方法通过体外冲击波碎石机建立Ⅱ型肺泡上皮细胞冲击波损伤模型,用CCK-8检测细胞活性,ELISA检测细胞上清中IL-6和IL-8含量;用0.5μg/mL的胡桃醌预处理细胞,再次经过冲击波处理后,CCK-8检测细胞活力,ELISA检测IL-6和IL-8的水平变化,Western blot检测caspase-3和caspase-9蛋白变化。结果CCK-8结果证实体外冲击波冲击1000次细胞活性降低至(49.2±3.04)%,伴有IL-6和IL-8表达升高,分别为(55.6±0.4)pg/mL和(1480.3±19.1)pg/mL,说明模型建造成功;用0.5μg/mL的胡桃醌预处理6 h后,再进行冲击波致伤,细胞活性升高至(70.1±3.1)%,IL-6、IL-8降低为(37.3±1.2)pg/mL和(1281.9±19.3)pg/mL,和冲击波损伤组相比差异有统计学意义(P<0.05)。Western blot结果显示,caspase-3和caspase-9的表达水平较冲击波损伤组降低(P<0.05)。结论胡桃醌能够减少冲击波导致的肺泡Ⅱ型上皮细胞的凋亡,降低炎症反应水平,从而发挥保护作用。

Objective waves induced injuries in typeⅡalveolar epithelial cells.MethodsThe injury of A549 cells caused by lithotripter shock wave(LSW)has been developed.CCK-8 was used to detect cell viability and ELISA was performed to detect the content of IL-6 and IL-8 in the supernatant.A549 cells were pretreated with Juglone(0.5μg/mL)for 6 hours followed by LSW for 1000 times,and the cell viability and the content of IL-6 and IL-8 in the supernatant were detected again,and the western blot analysis was applied to detect the protein expression levels of caspase-3 and caspase-9.Results viability of A549 cells to(49.2±3.04)%,while increasing the IL-6 and IL-8 level to(55.6±0.4)pg/mL and(1480.3±19.1)pg/mL respectively,which indicating that the shockwave injured cell model was established successfully.A549 cells were pretreated with Juglone(0.5μg/mL)for 6 hours before being exposed to LSW 1000 times,increasing cell viability to 70.13.1%while decreasing the content of IL-6 and IL-8 in the supernatant to(37.3±1.2)pg/mL and(1281.9±19.3)pg/mL,respectively.Compared with the shockwaves group,the difference was statistically significant(P<0.05).WB assay showed the decreased expression level of caspase-3 and caspase-9 following pre-treated with Juglone(P<0.05).Conclusion shock wave through reducing the apoptosis level and inhibiting inflammatory response.