Simulation for the smooth muscle layer of blood vessel plays a key role in tubular tissue engineering. However, fabrication of biocompatible tube with defined inner nano/micro-structure remains a challenge. Here, we show that a biocompatible polymer tube from poly(l-lactide) (PLLA) and polydimethylsiloxane (PDMS) can be prepared by using electrospinning technique, with assistance of rotating collector and parallel auxiliary electrode. The tube has circumferentially aligned PLLA fibers in the inner surface for cell growth regulation and has a PDMS coating for better compressive property. MTT assay showed the composite PLLA/PDMS tube was suitable for various cells growth. In vitro smooth muscle cells (SMCs) cultured in the tube showed that the aligned PLLA fibers could induce SMCs’ orientation, and different expression of α-SMA and OPN genes were observed on the aligned and random PLLA fibers, respectively. The successful fabrication of composite PLLA/PDMS tubular scaffold for regulating smooth muscle cells outgrowth has important implications for tissue-engineered blood vessels.