Abstract: OBJECTIVE To observe the efficiency of sanguinarine (SAN) acting on oxidative stress of H9c2 cell in vitro induced by lipopolysaccharide (LPS), and investigate its relationship with HO-1/NOX2 pathway. METHODS After H9c2 cells received intervention by different factors, cell growth was determined by CCK8 assay, reactive oxygen species (ROS) generation was detected by a Micro plate reader and Fluorescence microscopy, the mRNA expression of NOX2, P47PHOX, SOD1, SOD2 and HO-1 were determined by real time RT-PCR, MDA detection kit was determined to test the change of MDA. RESULTS SAN alone had no effect on cell activity; LPS reduced cell viability significantly, while co-treated the H9c2 cells with SAN and LPS could reverse this condition, improve cell activity and reduce ROS; the mRNA expression of NOX2, p47phox were increased and HO-1 decreased after LPS treated for 12 h, pre-treated with SAN, the mRNA expression of NOX2, p47phox were decreased and HO-1 was increased, while, pre-treated with SnPPIX and SAN could reverse this condition. SAN could increase the mRNA expression of SOD1 and SOD2, decrease the MDA level. CONCLUSION SAN inhibits the ROS generation induced by LPS via activate the HO-1/NOX2 pathway and also enhance ROS scavenging ability of H9c2 cell, and enhance the cell activity in the end.