Figure 4.

miR‐499 from injured NRCMs internalizes into RPMECs and worsens high glucose‐induced injury. A, Cartoon showing indirect co‐culture model of NRCMs and RPMECs using CdM. The CdM from NRCMs subjected to HX/R treatment significantly increased the expression levels of IL‐1α, ICAM‐1 and MCP‐1 both at mRNA and protein levels as evaluated by qRT‐PCR. B, (n = 3 and P < 0.05 for all) and ELISA. C, (n = 3 and P < 0.05 for all), which could be significantly blocked by AntagomiR‐499. D, CCK‐8 analysis showed that the CdM from NRCMs subjected to HX/R further decreased the cell viability induced by high glucose, which could be ameliorated by down‐regulation of the endogenous miR‐499 in NRCMs (n = 3 and P < 0.05 for all). E, ROS assay showed that the CdM from NRCMs subjected to HX/R further increased the ROS levels of RPMECs caused by high glucose, which could be partially inhibited by antagomiR‐499 (n = 3 and P < 0.05 for all). Experiments were performed three times in triplicate, and data are presented as ‘Mean ± SD’. The comparison between two groups was performed using paired or non‐paired Student's t test, with * denoting P < 0.05