Background and Aim: Type‐2 diabetes (T2D) is a highly prevalent, progressive disease constituting the major challenge to global health. MicroRNAs (miRNAs) play an important role in regulating physiological processes in diabetes through binding multiple target mRNAs, resulting in gene silencing. In the present study, the primary objective is to explore the role of miR221/222 and miR103/107 in diabetes.Materials and Methods: Mouse microvascular endothelial cells (MMECs) were cultured for 48 hours under conditions designed to mimic the mileu of T2D mice: normal glucose (NG=11mM) and high glucose (HG=40mM). The levels of miRNA221/222 and mir103/107 expression were analysed by real‐time PCR. MMECs were transfected with specific miR inhibitors/ mimics and then cells were exposed to normal/ high glucose in the presence or absence of metformin. The effects of miR expressions on Akt, P‐Akt, c‐kit, eNOS, P‐eNOS and Cav1 protein levels were determined through western blotting.Results: We found that exposure to high glucose increased the expression of miR‐221/222 but decreased the expression of miR‐103/107. Treatment with metformin partially restored HG‐induced expression of miRNAs towards that seen in control, NG, mileau. Furthermore, we observed that protein levels of total eNOS and P‐eNOS decreased in HG in comparison with NG. The eNOS/P‐eNOS protein levels were restored upon inhibition of miR221 but not with miR‐222, miR‐103 or miR‐107, thus indicating correlation of miR‐221 with eNOS signalling. Results were validated by using miR‐221 mimics (overexpression) and known target (c‐kit) of miR221.Conclusion: These findings suggest that miR221 may offer a new therapeutic strategy for treatment of endothelial dysfunction in diabetic patients or may work as a therapeutic modulator.