Overexpression of microRNA‐138 alleviates human coronary artery endothelial cell injury and inflammatory response by inhibiting the PI3K/Akt/eNOS pathway

摘要

This study aimed to investigate the role of miR‐138 in human coronary artery endothelial cell (HCAEC) injury and inflammatory response and the involvement of the PI3K/Akt/eNOS signalling pathway. Oxidized low‐density lipoprotein (OX‐LDL)‐induced HCAEC injury models were established and assigned to blank, miR‐138 mimic, miR‐138 inhibitor, LY294002 (an inhibitor of the PI3K/Akt/eNOS pathway), miR‐138 inhibitor + LY294002 and negative control (NC) groups. qRT‐PCR and Western blotting were performed to detect the miR‐138, PI3K, Akt and eNOS levels and the protein expressions of PI3K, Akt, style="fixed-case">eNOS, p‐Akt, p‐ style="fixed-case">eNOS, Bcl‐2, Bax and caspase‐3. style="fixed-case">ELISAs were employed to measure the expressions of style="fixed-case">TNF‐α, style="fixed-case">IL‐4, style="fixed-case">IL‐6, style="fixed-case">IL‐8, style="fixed-case">IL‐10 and nitric oxide ( style="fixed-case">NO) and the activities of lactate dehydrogenase ( style="fixed-case">LDH) and style="fixed-case">eNOS. style="fixed-case">MTT and flow cytometry were performed to assess the proliferation and apoptosis of style="fixed-case">HCAECs. Compared to the blank group, style="fixed-case">PI3K, Akt and style="fixed-case">eNOS were down‐regulated in the miR‐138 mimic and style="fixed-case">LY294002 groups but were up‐regulated in the miR‐138 inhibitor group. The miR‐138 mimic and style="fixed-case">LY294002 groups showed decreased concentrations of style="fixed-case">TNF‐α, style="fixed-case">IL‐6, style="fixed-case">IL‐8 and style="fixed-case">NO and reduced activities of style="fixed-case">LDH and style="fixed-case">eNOS, while opposite trends were observed in the miR‐138 inhibitor group. The concentrations of style="fixed-case">IL‐4 and style="fixed-case">IL‐10 increased in the miR‐138 mimic and style="fixed-case">LY294002 groups but decreased in the miR‐138 inhibitor group. The miR‐138 mimic and style="fixed-case">LY294002 groups had significantly decreased cell proliferation and increased cell apoptosis compared to the blank group. These findings indicate that up‐regulation of miR‐138 alleviates style="fixed-case">HCAEC injury and inflammatory response by inhibiting the style="fixed-case">PI3K/Akt/ style="fixed-case">eNOS signalling pathway.