莲心碱通过抑制细胞自噬减轻体外胰腺腺泡细胞急性炎症反应OACSTPCD

Objective To explore the protective role of liensinine(LSN)in a cellular model of cerulein-induced acute pancreatitis and its underlying mechanism.Methods Pancreatic acinar cells AR42J were divided into control group,model group,20 μmol/L LSN group,and 40 μmol/L LSN group.AR42J cells in model group were stimulated by cerulein,and AR42J cells in 20 μmol/L LSN group and 40 μmol/L LSN group were pretreated with LSN for 12 h followed by stimulation with cerulein.Protein levels of LC3,Beclin-1 and p62 were measured by Western blot.The levels of inflammatory cytokines including interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)were assessed by ELISA.Amylase level was analyzed by amylase assay kit.Cell survival rate was measured by Calcein AM cell viability assay kit.Cell injury was determined by lactic dehydrogenase(LDH)cytotoxicity assay kit.Cell apoptosis was detected by TUNEL assay.Results Compared with control group,LC3-Ⅱ/LC3-Ⅰ ratio and Beclin-1 protein level were significantly increased(P＜0.01),while p62 protein level was significantly decreased in model group(P＜0.01).Compared with model group,LC3-Ⅱ/LC3-Ⅰ ratio and Beclin-1 protein level were significantly decreased(P＜0.05),while p62 protein level was significantly increased in 20 μmol/L LSN group and 40 μmol/L LSN group(P＜0.05).Compared with 20 μmol/L LSN group,LC3-Ⅱ/LC3-Iratio and Beclin-1 protein level were significantly reduced(P＜0.05),while p62 protein level was significantly upregulated in 40 μmol/L LSN group(P＜0.05).Compared with control group,the levels of IL-1β,IL-6 and TNF-α were significantly increased(P＜0.01),the amylase level was significantly increased(P＜0.01),the cell survival rate was significantly reduced(P＜0.01),LDH activity was significantly elevated(P＜0.01),and the cell apoptosis rate was increased in model group(P＜0.01).Compared with model group,the levels of IL-1 β,IL-6 and TNF-α were significantly decreased(P＜0.05),the amylase level was significantly decreased(P＜0.05),the cell survival rate was significantly improved(P＜0.05),LDH activity was significantly reduced(P＜0.05),and the cell apoptosis rate was significantly decreased in 20 μmol/L LSN group and 40 μmol/L LSN group(P＜0.05).Compared with 20 μmol/L LSN group,the levels of IL-1β,IL-6 and TNF-α were significantly decreased(P＜0.05),the amylase level was significantly reduced(P＜0.05),the cell survival rate was significantly increased(P＜0.05),LDH activity was significantly decreased(P＜0.05),and the cell apoptosis rate was significantly decreased in 40 μmol/L LSN group(P＜0.05).Conclusion LSN can effectively protect against cerulein-induced injury of pancreatic acinar cells possibly through the suppression of autophagy.