Abstract: Objective Nitric oxide (NO) and soluble guanylate cyclase (sGC) are both oxidized under the condition of diabetes, resulting in abnormal conduction of the nitric oxide/soluble guanylate cyclase/cyclic guanosine monophosphate (NO-sGC-cGMP) signaling pathway, which is the important pathological basis of diabetic endothelial dysfunction. Cinaciguat (CIN) is a soluble guanylate cyclase (sGC) activator that can protect endothelial cells by activating sGC. In the present study, we investigated the effect of Cinacigua on human umbilical vein endothelial cells (HUVECs) treated with high glucose in vitro. Methods The cultured HUVECs were divided into the normal control group, high glucose group and CIN group, with 6 cases in each group. High glucose group and CIN group were treated with high glucose (33.3 mmol/L), and the CIN group was also given CIN. The levels of NO and nitric oxide synthase (NOS) were measured after the intervention with 6 h, 24 h, 48 h. Forty-eight h after the intervention, mRNA expression eNOS and iNOS were examined by using RT-PCR, and the expression of ICAM-1 and VCAM-1 were examined by using Western blotting. Results Compared with normal control group, the levels of NO and NOS in the high glucose group increased at early stage and decreased at later stage (P<0.05), but CIN could balance the levels of NO and NOS (P<0.05). In the same time, the expressions of eNOS mRNA was down-regulated, In addition, ICAM-1 and VCAM-1 were up-regulated in the state of high glucose, but the level of eNOS mRNA in CIN group was higher than that in high glucose group, and the level of iNOS mRNA, ICAM-1 and VCAM-1 decreased obviously (P<0.05). Conclusion CIN can balance the activity of NOS, adjust the expression of NO, ICAM-1 and VCAM-1 of HUVECs induced by high glucose, and improve function of injured HUVECs.