Knockdown of miR-423-5p simultaneously upgrades the eNOS and VEGFa pathways in ADSCs and improves erectile function in diabetic rats

Abstract Background Erectile dysfunction (ED) is both physical and psychological problem to men. The treatment is still unsatisfied. Modified adipose derived stem cells (ADSCs) may be a good therapy. This study aim to investigate if knockout of miR-423-5p in ADSCs alleviates damage to endothelial cells in high glucose conditions and improves erectile function in diabetic rats. Methods MiR-423-5p was knocked down in ADSCs. Then, ADSCs, NC-miR-ADSCs, and miR-ADSCs were co-cultured with human umbilical vein endothelial cells (HUVECs). Normal glucose and high glucose media were supplemented. Afterwards, the supernatant and HUVECs were collected for assessment of eNOS and VEGFa expression, cell proliferation, and apoptosis. Fifteen male Sprague-Dawley (SD) rats aged 12 weeks were induced to develop diabetes mellitus by intraperitoneal injection with streptozocin (STZ), and five healthy SD rats were used as normal controls. Eight weeks after developing diabetes, the rats received ADSCs and miR-ADSCs via injection into the corpora cavernosa, whereas normal controls and DM controls were injected with saline. Erectile function and histological assessment of penile tissues were performed eight weeks after injection. Results MiR-423-5p was successfully inhibited in ADSCs, and HUVECs co-cultured with ADSCs or miR-ADSCs exhibited higher eNOS and VEGFa protein expression levels compared to DM groups (P<0.05). MiR-ADSCs enhanced HUVEC proliferation compared to the ADSCs and NC-miR-ADSCs in both normal and high glucose conditions (P<0.01). Lower apoptotic rates were observed when HUVECs were co-cultured with miR-ADSCs, compared to ADSCs and NC-miR-ADSCs (P<0.05). The ICP/MAP indicated that erectile function was severely impaired in the DM rats compared with the normal group. Injection of ADSCs and miR-ADSCs improved erectile function (P<0.01) significantly and was associated with the overexpression of eNOS and VEGFa. Masson trichrome stain showed less fibrosis formation in the miR-ADSC group than the DM group. Conclusions MiR-423-5p knockdown in ADSCs ameliorated high glucose-mediated damage to HUVECs and improved erectile function in DM rats by inducing eNOS and VEGFa overexpression, indicating that miR-423-5p may be a potential target in the treatment of DM erectile dysfunction (DMED).