Abstract: Objective To investigate the role of resveratrol(RES) in ameliorating 4-hydroperoxy cyclophosphamide(4-HC)-induced oxidative stress damage in human granulosa cells(GCs). 
Methods The GCs were collected from patients undergoing in vitro fertilization and embryo transfer(IVF-ET) for in vitro culture. Hematoxylin-eosin staining(HE) and immunochemical staining were performed to identify GCs. Cell Counting Kit-8(CCK-8) was used to detect the toxicity of 4-HC-induced GCs and the improvement effect of RES on the toxicity of 4-HC-induced GCs. The GCs were divided into three groups: the control group, 4-HC group and RES+4-HC group. The CCK-8 was used to measure survival rate of GCs, and DCFH-DA fluorescent probe was used to detect reactive oxygen species(ROS) in GCs. Water-soluble tetrazolium-1(WST-1) was used to detect superoxide dismutase(SOD), and ammonium molybdate method was used to detect catalase(CAT). The protein expression of Silent Information Regulator 2-related enzyme 1(SIRT1) was determined by Western Blot. 
Results GCs were treated with 4-HC( 0, 1.0, 2.5, 5.0, 7.5 μmol/L). CCK-8 results showed the survival rate of GCs decreased with the increase of 4-HC, which showed significant differences, as compared with the control group(P＜0.05). 2.5 μmol/L 4-HC was selected as the subsequent concentration of 4-HC model. RES(0, 5, 10, 25, 50, 75 μmol/L) was used to treat GCs for 2 h and then added with 2.5 μmol/L 4-HC. The results showed that RES improved the survival rate of GCs, suggesting significant differences compared with the 4-HC group(P＜0.05). 50.0 μmol/L RES was selected as the subsequent improvement concentration. The detection results of GCs after treatment in the three groups showed that survival rate of GCs, SOD and CAT decreased, and ROS increased in 4-HC group, compared with CON group(P＜0.05). Compared with the 4-HC group, the survival rate of GCs, SOD and CAT in the RES+4-HC group significantly increased, and the ROS significantly decreased. Western blot showed that SIRT1 in the RES+4-HC group significantly increased, suggesting significant difference, compared with the other two groups(P＜0.05). 
Conclusion RES possibly ameliorates 4-HC-induced GCs damage by increasing SIRT1.

Key words: resveratrol, cyclophosphamide, fertilization in vitro