Effect Of Resveratrol Oil Expression Of Lipopolysaccharide-induced MicroRNA-146a In Alveolar Macrophages

Objective：To observe the effect of resveratrol on expression of lipopolysaccharide(LPS)-induced microRNA-146a(miR-146a)in alveolar macrophages, and investigateregulative effect and potential mechanism of miR-146a in alveolar macrophagesinflammatory responses, then provide a novelty approach to treat sepsis-inducedlung injury.Methods：Rat alveolar macrophages(NR8383)were seeded at2×106cell/pore in6wellplates in vitro and incubated for90min, then NR8383were randomly divided intofour groups: control group that stimulated with phosphate buffer solution (PBS);LPS-stimulated group that stimulated with1μg/ml of LPS; resveratrol-treated groupthat treated cells with different concentrations resveratrol (1μg/ml or10μg/ml) for30min, then stimulated with1μg/ml of LPS. After incubation for6h, observed thechange of cellular morphology and harvested the cells and collected the supernatant.The expression of TNF-α mRNA and miR-146a of cells were detected by Real-timequantitative PCR and the production of TNF-α protein in the supernatant of cellswere assayed by enzyme-linked immunosorbent assay (ELISA).Results：1． After incubation for6h, LPS inhibited cell growth and cell adhesion, alsoinduced one part of cells death, but treated cells with resveratrol could attenuate thecellular injury and protect cell growth and cell adhesion, the protection had beendose dependent;2． After incubation for6h, compared with control group, the expression ofTNF-α mRNA was significantly increased (50.05±8.08fold, P<0.01), the productionof TNF-α in the supernatant of cells was significantly increased (from26.15±13.43pg/ml to644.2±36.82pg/ml, P<0.01) in LPS-stimulated group, but inresveratrol-treated group (1μg/ml and10μg/ml), the expression of TNF-α mRNA were increased less (29.64±8.39fold, P<0.01) and (13.54±4.50fold, P<0.01), theproduction of TNF-α in the supernatant of cells were increased less(447.7±41.48pg/ml, P<0.01)and(345.0±42.54pg/ml, P<0.01);3． After incubation for6h, compared with control group, the expression ofmiR-146a was increased (5.92±1.57, P<0.01) in LPS-stimulated group, but inresveratrol-treated group (1μg/ml and10μg/ml), the expression of miR-146a weresignificantly increased (8.67±1.18fold, P＜0.01) and (11.17±1.40fold, P＜0.01);Conclusion：Treating NR8383with resveratrol, could attenuate LPS-induced cellular injuryand inhibit expression of the inflammatory factor: TNF-α, also up-regulate theexpression of miR-146a, so it could be supposed that resveratrol had been blockedrelease of inflammatory mediators in alveolar macrophage inflammatory response byup-regulating the expression of miR-146a inhibiting TLRs/NF-κB pathway.