Objective To explore whether sphingosine 1-phosphate ( S1P) is involved in pancreatic fibrosis in KkAy type 2 diabetic mice, and whether this process is related to transforming growth factor-β1 (TGF-β1). Methods Real-time (RT)-PCR was used to detect the level of sphingosine kinase 1 (SphK1), TGF-β1 and smooth muscle actin α (α-SMA), which were measured as an index that reflects the activation of pancreatic stellate cells in the pancreas in KkAy type 2 diabetic mice and C57 mice. The main components of the extracellular matrix, collagen α1( I) [Col α1( I)] and collagen α1(III) [Col α1(III)], were also measured by Real-time RT-PCR. Results SphK1, TGF-β1, α-SMA, Col α1 (I) and Col α1( III) were overexpressed in the pancreas in KkAy type 2 diabetic mice. There was a significant positive correlation between SphK1 and TGF-β1 in pancreatic tissue of KkAy type 2 diabetic mice. Conclusions S1P may be involved in the development of pancreatic fibrosis in KkAy type 2 diabetic mice, and this process may be related to TGF-β1.