A multi-layer interconnection structure is a basic component of electronic devices, and printing of the multi-layer interconnection structure is the key process in printed electronics. In this work, electrohydrodynamic direct-writing (EDW) is utilized to print the conductor-insulator--conductor multi-layer ~nterconne^ction structure. Silver ink is chosen to print the conductor pattern, and a polyvinylpyrrolidone (PVP) solution is util^zed to f^bricate the insulator layer between the bottom and top conductor patterns. The influences of EDW process parameters on the line width of the printed conductor and insulator patterns are studied systematically. The obtained ~es^l~s show that the line width of the printed structure increases with the increase of the flow rate, but decreases with the increase of applied voltage and PVP content in the solution. The average resistivity values of the bottom and top silver conductor tracks are determined to be 1.34 × 10-7 Ω.m and 1.39×10-7 Ω.m, respectively. The printed PVP layer between the two conductor tracks is well insulated, which can meet the insulation requirement of the electronic devices. This study offers an alternative, fast, and cost-effective method of fabricating conductor-insulator-conductor multi-layer interconnections in the electronic industry.