Aim To study the protective effect and possible mechanism of Brazilin on high glucose induced vascular endothelial cell injury, and to provide experimental basis for the treatment of diabetic macrovascular disease with traditional Chinese medicine hematoxylin. Methods Human umbilical vein endothelial cells (HUVEC) were cultured, the concentration of Brazilin was determined by MTT method, high glucose induced injury after joining Brazilin, autophagy body was observed by transmission electron microscopy (TEM), tubule formation test was used to observe angiogenesis, the protein expression of autophagy related genes BECLIN1, LC3-Ⅱ and p62 was detected by Western blot. Results The concentration of Brazilin was 15 mg/L. Transmission electron microscopy showed that the number of autophagic cells in high glucose treatment group was less than that in normal group, but the number of autophagy in the cells of Brazilin group was higher than that in normal group and high glucose treatment group. The expression of BECLIN1 and LC3-Ⅱ protein in high glucose treated group was lower than that in normal group, the expression of BECLIN1 and LC3-Ⅱ protein in Brazilin group was higher than that in high glucose treatment group (P＜0.05); the expression of p62 protein in high glucose treated group was higher than that in normal group, the expression of p62 protein in Brazilin group was lower than that in normal group and high glucose treatment group (P＜0.05). Conclusion Brazilin may play a role of protection in high glucose induced vascular endothelial cell injury through regulating autophagy.