Ang 1-7对雨蛙素刺激的胰腺腺泡AR42J细胞p38 MAPK/NF-κB信号通路的影响

Effect of Ang 1-7 on the p38 MAPK/NF-κB signaling pathway in caerulein- stimulated pancreatic acinar AR42J cell

目的探讨血管紧张素1-7(Ang 1-7)干预雨蛙素(caerulein,CAE)刺激的胰腺腺泡细胞AR42J后,p38MAPK/NF-κB信号蛋白的表达变化。方法将AR42J细胞随机分为3组:对照组、雨蛙素组(10 nmol/L,CAE组)、Ang1-7组(10-7mol/L、10-6mol/L、10-5mol/L Ang 1-7+10 nmol/L雨蛙素)。对照组为正常生长的AR42J细胞,CAE组用10 nmol/L的雨蛙素刺激AR42J于2 h、6 h、12 h、24 h及48 h收集细胞,Ang 1-7组为不同浓度Ang 1-7作用30 min后再用雨蛙素刺激24 h收集细胞,Western blot技术检测各组细胞中ACE2、Mas受体、p38 MAPK、p-p38 MAPK及NF-κB的蛋白表达。结果 AR42J细胞中可见ACE2-Ang 1-7-Mas受体轴表达;与对照组相比,CAE组ACE2和Mas受体的表达呈先增多后减少的趋势,均于24 h达高峰(P<0.05);p38 MAPK、p-p38 MAPK及NF-κB的表达亦是24 h达峰值(P<0.05);Ang 1-7(10-7mol/L、10-6mol/L及10-5mol/L)上调Mas受体的表达,下调p38 MAPK、p-p38MAPK、NF-κB蛋白的表达,Ang 1-7的浓度为10-5mol/L时,后三者的表达水平与CAE组比较,差异均具有统计学意义(P<0.05)。结论雨蛙素刺激的AR42J细胞中,Ang 1-7可通过Mas受体抑制p38MAPK/NF-κB信号通路的活性。

Objective To study the effect of Angiotensin 1-7（Ang 1-7）on the p38 MAPK/NF-κB signaling pathway in caerulein-stimulated pancreatic acinar AR42J cell. Methods AR42J cells were randomly divided into 3 groups,including control group,caerulein group （10 nmol/L,CAE）and Ang 1-7 group（10 ^-7、10^ -6、10 ^-5 mol/L Ang 1^-7 ＋ 10 nmol/L CAE）. The control group was AR42J cell growing normally. AR42J cells were stimulated with 10 nmol/L caerulein for varying time periods and collected at 2 h,6 h,12 h,24 h and 48 h post stim-ulation. Before the stimulation of caerulein for 24 h,the Ang 1-7 group cells were mock pretreated or pretreated with different concentrations of Ang 1-7 for 30 min and 24 h post the treatments of caerulein,each group ACE2,Mas receptor,p38MAPK,p-p38MAPK,and NF-κB ex-pression levels were evaluated by Western Blotting. Results ACE2-Ang 1-7-Mas axis existed in AR42J cell. Compared with control group, the expression of ACE2 and Mas receptor of CAE group increased and then decreased ,both peaked at 24 h（ P ＆lt;0. 05）. The p38 MAPK,p-p38 MAPK and NF-κB were also peaked at 24 h（ P ＆lt;0. 05）. Ang 1-7 group（10 -7 mol/L,10 -6 mol/L and 10 -5 mol/L）up-regulated the expression of Mas receptor,down-regulated p38 MAPK,p-p38 MAPK,NF-κB expression. When the concentration of Ang 1 -7 was 10 -5 mol/L,the latter three protein levels were statistically significant（ P ＆lt;0. 05）compared with CAE group. Conclusion In the caerulein-stimulated AR42J cell,Ang（1-7）may inhibits p38MAPK/ NF-κB signaling pathway through Mas receptor.