Effect of resveratrol on expression of lipopolysaccharide-induced microRNA-146a in alveolar macrophages
Zhenguo Zeng, Qiang Shao, Chengzhi Ding, Qing Cheng, Fen Liu, Yian Zhan
Abstract
Zhenguo Zeng, Qiang Shao, Chengzhi Ding, Qing Cheng, Fen Liu, Yian Zhan
Abstract
Objective To observe the effect of resveratrol on expression of lipopolysaccharide (LPS) -induced microRNA-146a (miR-146a) in alveolar macrophages.Methods Rat alveolar macrophages (NR8383) were seeded at 2 × 106cell/pore in 6 well plates in vitro and incubated for 90 min.NR8383 were randomly divided into four groups:control group,NR8383 stimulated with phosphate buffer solution (PBS) ; LPS-stimulated group,NR8383 stimulated with 1 μg/mL of LPS ; resveratrol-treated group,NR8383 treated with different concentrations of resveratrol (1 μg/mL or 10 μg/mL) for 30 min,then stimulated with 1 μg/ mL of LPS.After incubation for 6 h,the cells were harvested and the supernatant of cell culture medium was collected.The expression of miR-146a of cells was detected by using fluoroilluminance real-time quantitative PCR device,and the levels of TNF-α protein in the supernatant were assayed by using enzyme-linked immunosorbent assay (ELISA).Results Compared with control group,the expression of miR-146a and the level of TNF-oα were significantly increased in LPS stimulated group (miR-146a' s expression folds:5.92 ±l.57vs 1.03±0.58,P<0.01; TNF-α:644.2±36.82 pg/mL vs.26.15 ±13.43 pg/mL,P<0.01).Compared with LPS-stimulated group,the expressions of miR-146a were significantly increased in resveratrol-treated groups (1 μg/mL and 10 μg/mL) (miR-146a' s expression folds:8.67 ± 1.18,11.17± 1.40 vs.5.92 ± 1.57,both P < 0.01),but the level of TNF-oα in the supernatant was significantly reduced (TNF-α:447.7 ± 41.48 pg/mL,345.0 ± 42.54 pg/mL vs.644.2 ± 36.82 pg/mL,both P < 0.01).Compared with the low concentration of group (1 μg/mL),the expression of miR-146a were significantly increased and the production of TNF-α in the supernatant of cells was significantly reduced in high concentration of resveratrol group (10 μg/mL) (P < 0.01).Conclusions Resveratrol could upregulate the expression of miR-146a in alveolar macrophage inflammatory response,suggesting miR-146a involved in the anti-inflammtory processes with resveratrol treatment. Key words: Resveratrol; MicroRNA-146a; Alveolar macrophages; Tumor necrosis factor-α
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Objective To observe the effect of resveratrol on expression of lipopolysaccharide (LPS) -induced microRNA-146a (miR-146a) in alveolar macrophages.Methods Rat alveolar macrophages (NR8383) were seeded at 2 × 106cell/pore in 6 well plates in vitro and incubated for 90 min.NR8383 were randomly divided into four groups:control group,NR8383 stimulated with phosphate buffer solution (PBS) ; LPS-stimulated group,NR8383 stimulated with 1 μg/mL of LPS ; resveratrol-treated group,NR8383 treated with different concentrations of resveratrol (1 μg/mL or 10 μg/mL) for 30 min,then stimulated with 1 μg/ mL of LPS.After incubation for 6 h,the cells were harvested and the supernatant of cell culture medium was collected.The expression of miR-146a of cells was detected by using fluoroilluminance real-time quantitative PCR device,and the levels of TNF-α protein in the supernatant were assayed by using enzyme-linked immunosorbent assay (ELISA).Results Compared with control group,the expression of miR-146a and the level of TNF-oα were significantly increased in LPS stimulated group (miR-146a' s expression folds:5.92 ±l.57vs 1.03±0.58,P<0.01; TNF-α:644.2±36.82 pg/mL vs.26.15 ±13.43 pg/mL,P<0.01).Compared with LPS-stimulated group,the expressions of miR-146a were significantly increased in resveratrol-treated groups (1 μg/mL and 10 μg/mL) (miR-146a' s expression folds:8.67 ± 1.18,11.17± 1.40 vs.5.92 ± 1.57,both P < 0.01),but the level of TNF-oα in the supernatant was significantly reduced (TNF-α:447.7 ± 41.48 pg/mL,345.0 ± 42.54 pg/mL vs.644.2 ± 36.82 pg/mL,both P < 0.01).Compared with the low concentration of group (1 μg/mL),the expression of miR-146a were significantly increased and the production of TNF-α in the supernatant of cells was significantly reduced in high concentration of resveratrol group (10 μg/mL) (P < 0.01).Conclusions Resveratrol could upregulate the expression of miR-146a in alveolar macrophage inflammatory response,suggesting miR-146a involved in the anti-inflammtory processes with resveratrol treatment. Key words: Resveratrol; MicroRNA-146a; Alveolar macrophages; Tumor necrosis factor-α
Key concepts: Resveratrol, Lipopolysaccharide, Alveolar macrophage, Incubation, Molecular biology, In vitro, Medicine, Tumor necrosis factor alpha