Effects of spautin-1 on autophagy and apoptosis of acute pancreatitis cell model induced by cerulein / 中华胰腺病杂志

ABSTRACT

Objective To investigate the potential of Spautin-1 to treat acute pancreatitis by inhibiting impaired autophagy and promoting apoptosis using an acute pancreatitis cell model induced by cerulein in vitro.Methods Pancreatic acinar cells AR42J were treated with sterile saline,100 nmol/L cerulein,and 100 nmol/L cerulein combined with 10 μmol/L spautin-1 for 24 h,respectively,and then western blot was used to detect the expression of LC3,p62 and Beclin1 in the cells to reflect autophagy;the trypsin activity in cells was detected by BGAM.Flow cytometry was used to detect the apoptosis;LDH cytotoxicity assay kit was used to detect cell necrosis.Results The expression of LC3 Ⅰ (0 h-24 h4.32±0.46,4.68±0.41,5.22± 0.38,5.88 ±0.63),LC3 Ⅱ (0 h-24 h0.36 ±0.02,0.64 ±0.05,0.93 ±0.08,2.43 ±0.23) and p62 (0 h-24 h0.24 ± 0.01,0.22 ± 0.02,0.84 ± 0.09,1.25 ± 0.13) was increased in AR42J cell model treated with cerulein (P ＜ 0.05),indicated that cells undergo autophagy,but autophagy flux is blocked.The expression of LC3 Ⅰ (0.65 ± 0.06 vs 0.24 ± 0.01),LC3 Ⅱ (1.26 ± 0.15 vs 0.71 ± 0.08) and p62 (1.06 ± 0.09 vs 0.56 ± 0.06) were decreased after incubated with spautin-1,with the decreased trypsin activity (1.65 ±0.18 vs 1.13 ±0.14),increased apoptosis (6.58 ±4.01 vs 23.64 ±2.12) (P ＜0.05) and reduced cellnecrosis (27.58± 3.46vs7.64 ± 2.12) (P＜0.05).Conclusions Inthe AR42 Jcellmodel ofacute pancreatitis induced by cerulein in vitro,spautin-1 can inhibit impaired autophagy induced by cerulein,decrease trypsin activity,increase apoptosis and reduce cell necrosis to protect pancreatic cells from damage,which has a certain value for remissing and even curing acute pancreatitis.