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02 June 2016 : Animal Research  

MicroRNA-219-5p Inhibits Morphine-Induced Apoptosis by Targeting Key Cell Cycle Regulator WEE1

Wei LouABCDEFG, Xingwang ZhangBCDE, Xiao-Ying HuABDE, Ai-Rong HuABDE

DOI: 10.12659/MSM.895439

Med Sci Monit 2016; 22:1872-1879

Abstract

BACKGROUND: To identify the effects of microRNA (miR)-219-5p on morphine-induced apoptosis by targeting WEE1.

MATERIAL AND METHODS: Forty Balb/C mice (Toll-like receptor 9, TLR9 knockout) were randomly allocated to the experimental and control groups (20 in each group). The baseline miR-219-5p expression was detected using quantitative real-time PCR (qRT-PCR). After morphine was injected at 6 h on the 2nd and 6th days, experimental and control groups received miR-219-5p mimics or miRNA-negative control (NC), respectively, compound injection. Tissues and cells were later obtained from subjects in each group separately after mice were killed. TUNEL assay was used to investigate apoptosis in both groups. RAW264.7 cells were treated with miR-219-5p mimics and controls, respectively. After 24 h, 10 μM of morphine was added at 24 h. Cell apoptosis was assessed by flow cytometer. The WEE1 and Phospho-cdc2 (Tyr15) expressions were examined by Western blotting.

RESULTS: MiR-219-5p expression in the experimental group was significantly lower than that in the control group (P<0.05). Mice injected with miR-219-5p mimic experienced an evident increase in apoptosis rate compared with the control group (P<0.05). The miR-219-5p NC group and the morphine group both presented an elevated apoptosis rate compared with the blank control group (both, P<0.05). The apoptosis rate in the miR-219-5p mimic group was 10.06%, remarkably lower than in the miR-219-5p NC group and blank control group (both P<0.05). WEE1 and Tyr15 protein expressions in the miR-219-5p NC group and morphine group were obviously stronger than those in the blank control group (all P<0.05). In the miR-219-5p mimic group, WEE1 and Tyr15 protein expressions were significantly lower compared with those in the miR-219-5p NC group and morphine group (all P<0.05).

CONCLUSIONS: Morphine significantly downregulated the expression of miRNA-219-5p, which targets WEE1 to suppress Tyr15 expressions and activate Cdc2, thus inhibiting the morphine-induced macrophage apoptosis.

Keywords: Apoptosis - genetics, Cell Cycle - genetics, Cell Cycle Proteins - metabolism, Down-Regulation - drug effects, Macrophage Activation - genetics, Macrophages, Peritoneal - physiology, Mice, Knockout, Morphine - pharmacology, Nuclear Proteins - metabolism, Protein-Tyrosine Kinases - metabolism, RAW 264.7 Cells, Random Allocation, Toll-Like Receptor 9 - metabolism

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750