title: Overexpression of microRNA-138 alleviates human coronary artery endothelial cell injury and inflammatory response by inhibiting the PI3K/Akt/eNOS pathway.
abstract: 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, eNOS, p-Akt, p-eNOS, Bcl-2, Bax and caspase-3. ELISAs were employed to measure the expressions of TNF-α, IL-4, IL-6, IL-8, IL-10 and nitric oxide (NO) and the activities of lactate dehydrogenase (LDH) and eNOS. MTT and flow cytometry were performed to assess the proliferation and apoptosis of HCAECs. Compared to the blank group, PI3K, Akt and eNOS were down-regulated in the miR-138 mimic and LY294002 groups but were up-regulated in the miR-138 inhibitor group. The miR-138 mimic and LY294002 groups showed decreased concentrations of TNF-α, IL-6, IL-8 and NO and reduced activities of LDH and eNOS, while opposite trends were observed in the miR-138 inhibitor group. The concentrations of IL-4 and IL-10 increased in the miR-138 mimic and LY294002 groups but decreased in the miR-138 inhibitor group. The miR-138 mimic and 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 HCAEC injury and inflammatory response by inhibiting the PI3K/Akt/eNOS signalling pathway.
authors.affiliation: Department of Cardiac Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Jing-Bo
authors.initials: JB
authors.lastname: Li
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Hai-Yang
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Yao
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Qing-Feng
authors.initials: QF
authors.lastname: Sun
authors.affiliation: Department of Cardiac Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Zong-Hong
authors.initials: ZH
authors.lastname: Liu
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Si-Qi
authors.initials: SQ
authors.lastname: Liu
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Zhuang
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Yun-Peng
authors.initials: YP
authors.lastname: Wang
authors.affiliation: Department of Cardiac Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Hong-Yu
authors.initials: HY
authors.lastname: Liu
mesh_terms: D000070416:Antagomirs / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D007378:Interleukins / Q000235:genetics / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics* / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000070416:Antagomirs; D053148:Caspase 3; D002867:Chromones; D007378:Interleukins; D008077:Lipoproteins, LDL; D035683:MicroRNAs; D009025:Morpholines; D052250:Nitric Oxide Synthase Type III; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D019253:Proto-Oncogene Proteins c-bcl-2; D014409:Tumor Necrosis Factor-alpha; D051028:bcl-2-Associated X Protein; C000595707:BCL2 protein, human; C546110:MIRN138 microRNA, human; C099252:oxidized low density lipoprotein; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C496314:NOS3 protein, human; C505407:CASP3 protein, human
keywords: PI3K/Akt/eNOS signalling pathway; human coronary artery endothelial cells; inflammatory response; microRNA-138