Abstract: Objective To probe the molecular mechanism and effect of oxymatrine (OM) on the secretion of extracellular matrix (ECM) in rat pancreatic stellate cell line LTC-14 induced by TGF-β1. Methods LTC-14 cells were divided into control group (LTC-14 cells in normal culture), TGF-β1 group (LTC-14 cells stimulated by 10 μg/L TGF-β1 for 12 h), TGF-β1+OM group (LTC-14 cells pretreated with 1 g/L OM for 30 min before TGF-β1 stimulation) and TGF-β1+ SiZNF850 group (LTC-14 cells were transiently transfected with ShRNA-ZNF580 plasmids for 24 h before TGF - β1 stimulation). After cultivation, cell supernatant was collected and total RNA and total protein were extracted. The expressions of Smad2, Smad3, ZNF580, α - SMA, MMP2 and TIMP1 were detected by Real-time PCR and Western blot assay. The secretion of ECM components, Col-I, Col-III, FN, TNF-α and IL-1β were detected by ELISA. Results The mRNA and protein expressions of Smad2, Smad3, ZNF580 and α-SMA were increased (P＜0.05), the expression ratio of MMP2/TIMP1 was down-regulated (P＜0.05), and the secretion of ECM components was increased in LTC-14 cells induced by TGF-β1 (P＜0.05). However, after OM intervention or ZNF580 gene knockdown, the mRNA and protein expressions of Smad2 and Smad3 were suppressed (P＜0.05), the mRNA and protein expressions of ZNF580 decreased, the expression ratio of MMP-2/ TIMP1 was up-regulated (P＜0.05), and the secretion of ECM components was reduced (P＜0.05). Conclusion Oxymatrine can reduce ECM secretion, increase ECM degradation and alleviate pancreatic fibrosis by inhibiting TGF- β1/ Smads/ZNF580 signaling pathway and blocking the activation of pancreatic stellate cells. ZNF580 may be a molecular target of OM in reducing pancreatic fibrosis.

Key words: oxymatrine, extracellular matrix, transforming growth factor beta 1, ZNF580, pancreatic stellate cell