基础医学与临床 ›› 2018, Vol. 38 ›› Issue (8): 1080-1087.

• 研究论文 • 上一篇    下一篇

NR5A2减弱雨蛙素诱导大鼠胰腺腺泡细胞的炎性反应

徐志刚1,郑帅1,杨泽冉2,陈雪3,肖鲁瑶1,孙亚梅3,张杰1   

  1. 1. 首都医科大学附属北京安贞医院
    2. 首都医科大学附属北京友谊医院
    3. 首都医科大学附属北京安贞医院消化内科
  • 收稿日期:2018-04-18 修回日期:2018-06-19 出版日期:2018-08-05 发布日期:2018-07-24
  • 通讯作者: 张杰 E-mail:zhangjie4155@sina.com
  • 基金资助:
    NR5A2对急性胰腺炎腺泡细胞损伤修复的影响及其机制研究

NR5A2 attenuates cerulein-induced inflammatory response in pancreatic acinar cells

  • Received:2018-04-18 Revised:2018-06-19 Online:2018-08-05 Published:2018-07-24

摘要: 目的 探讨NR5A2对大鼠胰腺腺泡细胞系AR42J炎性反应的影响。方法用雨蛙素刺激AR42J细胞,建立胰腺炎细胞模型。分别用慢病毒和sh-RNA感染AR42J细胞,上调和下调细胞内的NR5A2,用Western blot检测NR5A2的含量。在此基础上,用流式细胞计量术检测细胞凋亡率,用ELISA检测细胞培养上清中炎性因子IL-1、TNF-α的水平。结果 用100 nmol/L雨蛙素刺激AR42J细胞,成功诱导炎性反应和细胞凋亡,刺激6 h凋亡率达到峰值;刺激24 h NR5A2含量降到最低点,刺激48 h基本恢复至正常水平。用慢病毒载体过表达NR5A2,再用雨蛙素刺激,过表达组的细胞凋亡率显著高于对照组(P<0.05);IL-1和TNF-α在过表达组均显著降低(P<0.01,P<0.05)。用shRNA沉默NR5A2,雨蛙素刺激后沉默组的细胞凋亡率显著低于对照组(P<0.05);IL-1和TNF-α在沉默组均显著升高(P<0.05,P<0.01)。在过表达NR5A2基础上沉默β-catenin,与对照组相比,凋亡率降低(P<0.05),IL-1和TNF-α显著增高(P<0.05,P<0.01)。结论 NR5A2能够促进炎性反应条件下胰腺腺泡细胞的凋亡,抑制炎性反应。β-catenin参与NR5A2在腺泡细胞炎性反应应激条件下的调节作用。

关键词: 急性胰腺炎, 雨蛙素, NR5A2, β-catenin, 炎性因子

Abstract: Objective To study the protective role of NR5A2 to acinar cells under acute pancreatitis. Methods Acute pancreatitis model was created by stimulating AR42J cells with cerulein. Virus vectors and shRNA were respectively used to upregulate or downregulate NR5A2 expression, and NR5A2 changes were verified by Western blot. Quantification of cell apoptosis was tested by flow cytometry. IL-1 and TNF-α expression were detected by ELISA. Results The apoptosis rate was significantly elevated under 100 nmol/L cerulein until the peak at 6 hours, meanwhile NR5A2 level was decreased until 24 h stimulation, and then gradually recovered to basal level after 48 h. NR5A2 overexpression further increased the apoptosis rate under cerulein stimulation (P<0.05), and reduced IL-1 and TNF-α levels (P<0.01, P<0.05). On the other hand, NR5A2 inhibition by shRNA led to decreased apoptosis rate (P<0.05) and increased IL-1 and TNF-α levels (P<0.05, P<0.01).Furthermore, silencing β-catenin attenuated the effects of NR5A2-overexpression, that the apoptosis rate was significantly reduced (P<0.05), while IL-1 and TNF-α levels were significantly increased (P<0.05, P<0.01). Conclusions NR5A2 induces apoptosis and inhibites inflammatory reaction in AR42J cells under cerulein stimulation, and such effects are mediated by β-catenin.

Key words: acute pancreatitis, cerulein, NR5A2, β-catenin, inflammatory factors