实用医学杂志 ›› 2021, Vol. 37 ›› Issue (19): 2453-2457.doi: 10.3969/j.issn.1006⁃5725.2021.19.004

• 基础研究 • 上一篇    下一篇

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

田林强1 来枫1 张玥1 王玉华2 翁孝刚1 董新文3 姚三巧3 任文杰   

  1. 新乡医学院第三附属医院 1 创伤与骨科研究所,2 泌尿外科(河南新乡 453003);3 新乡医学院公共卫生学院 (河南新乡 453003) 
  • 出版日期:2021-10-10 发布日期:2021-10-10
  • 通讯作者: 任文杰 E⁃mail:xxmu_rwj@163.com
  • 基金资助:
    国家自然科学基金-河南省联合基金重点项目(编号:U1904209);河南省科技攻关项目(编号:202102310435)

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

TIAN Lin⁃ qiang*,LAI Feng,ZHANG Yue,WANG Yuhua,WENG Xiaogang,DONG Xinwen,YAO Sanqiao,REN Wen⁃ jie.   

  1. Institute of Trauma and Orthopedics,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,China

  • Online:2021-10-10 Published:2021-10-10
  • Contact: REN Wenjie E⁃mail:xxmu_rwj@163.com

摘要:

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 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 IL⁃6和IL⁃8含量;用0.5 μg/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1 000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 1 480.3 ± 19.1)pg/mL,说明模型建造成功;用0.5 μg/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(1 281.9 ± 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。

关键词:

胡桃醌, 细胞凋亡, 炎症反应, 冲击波, Ⅱ型肺泡上皮细胞

Abstract:

Objective The aim of this study is to investigate the protection effect of Juglone in shock ⁃ waves induced injuries in type Ⅱ alveolar epithelial cells. Methods The 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(0.5 μg/mL)for 6 hours followed by LSW for 1 000 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 The results of CCK⁃8 assays showed that shockwave treatment significantly decreased the 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 (1 480.3 ± 19.1)pg/mL respectively,which indicating that the shockwave injured cell model was established successfully. A549 cells were pretreated with Juglone(0.5 μg/mL)for 6 hours before being exposed to LSW 1 000 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 Juglone plays protecting role in A549 cells caused by shock wave through reducing the apoptosis level and inhibiting inflammatory response.

Key words:

Juglone, apoptosis, inflammation, shockwave, alveolar epithelial cells type Ⅱ