AIM: To investigate the therapeutic effect of tetrandrine (Tet) on mouse in vivo and in vitro models of severe acute pancreatitis (SAP) and its mechanisms. METHODS: (1) The pancreatic tissues of C57BL/6 mice were freshly isolated to obtain pancreatic acinar cells (PACs). The cells were randomly divided into blank control (Ctrl) group, Tet group, injury [induced by palmitoleic acid (POA) or taurolithocholic acid sodium salt (TLCS)] groups and Tet treatment (Tet+POA or Tet+TLCS) groups. The Fluo-4 fluorescent probe was used to measure cytoplasmic calcium concentration. Propidium iodide staining was used to detect necrotic cells. The TMRM fluorescent probe was used to measure the changes of mitochondrial membrane potential. Reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe. (2) Thirty-five 8-week-old healthy male C57BL/6 mice were randomly selected and divided into Ctrl group, sham group, Tet group, fatty acid ethyl ester-induced acute pancreatitis (FAEE-AP) group, TLCS-induced acute pancreatitis (TLCS-AP) group, Tet+FAEE-AP group and Tet+TLCS-AP group, with 5 mice in each group. The morphological changes of the pancreas were observed using HE staining. The positive staining rates of C/EBP homologous protein (CHOP) in the pancreas were observed by immunohistochemistry. The expression of glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase (PERK), p-PERK and CHOP was detected by Western blot. Serum levels of amylase, interleukin-6( IL-6), malondialdehyde( MDA), glutathione peroxidase( GSH-Px), superoxide dismutase( SOD) and catalase (CAT) were assayed. Myeloperoxidase (MPO) activity was detected in mouse pancreatic and lung tissues. RESULTS: Treatment with Tet effectively inhibited POA- or TLCS-induced calcium overload and necrosis in PACs, attenuated the decrease in mitochondrial membrane potential, and reduced ROS production in vitro (P<0. 01). Compared with Ctrl group, the pancreatic pathological score, serum amylase activity, serum IL-6 level, and MPO activity in the pancreas and lung in vivo significantly increased in SAP groups( P<0. 01). However, the administration of Tet significantly alleviated the tissue damage and inflammatory response (P<0. 01). Compared with SAP groups, Tet reduced the expression of endoplasmic reticulum stress-related GRP78/PERK/CHOP signaling pathway (P<0. 01), decreased serum peroxide MDA, and increased antioxidants GSH-Px, SOD and CAT (P<0. 01). CONCLUSION: Treatment with Tet effectively attenuates SAP in mouse models by the inhibition of calcium overload, the alleviation of mitochondrial membrane potential decrease, and the reduction of endoplasmic reticulum stress response and oxidative stress damage to protect against the necrosis of PACs. [ABSTRACT FROM AUTHOR]