CAJ | 학술논문

目的观察白藜芦醇对脂多糖(LPS)诱导肺泡巨噬细胞微小RNA-146a(miR-146a)表达的影响.方法将体外培养的大鼠肺泡巨噬细胞(NR8383)按2×106个/孔接种于六孔板,细胞贴壁90 min后,分为磷酸盐缓冲液(PBS)对照组、终质量浓度LPS(1μg/mL)处理组、白藜芦醇(1 μg/mL)预处理30 min+终质量浓度LPS(1μg/mL)处理组、白藜芦醇(1 0μg/mL)预处理30 min+终质量浓度LPS(1μg/mL)处理组.细胞培养6h后,收集细胞和细胞培养上清液,采用实时荧光定量PCR检测细胞中miR-146a的表达变化,采用酶联免疫吸附实验(ELISA)检测细胞培养上清液中TNF-α蛋白的表达变化.结果与PBS对照组相比,LPS处理组细胞中miR-146a和TNF-α蛋白表达量均明显升高[miR-146a表达倍数(5.92±1.57)倍vs.(1.03±0.58)倍,P＜0.01;TNF-α质量浓度(644.2±36.82) pg/mL vs.(26.15±13.43)pg/mL,P＜0.01].与LPS处理组相比,白藜芦醇预处理组(1μg/mL和10 μg/mL)细胞中miR-146a表达量均明显升高[miR-146a表达倍数(8.67±1.18)倍、(11.17±1.40)倍vs.(5.92±1.57)倍,P＜0.01],TNF-α蛋白质量浓度均显著降低[TNF-α质量浓度(447.7±41.48) pg/mL、(345.0±42.54)pg/mL vs.(644.2±36.82)pg/mL,P＜0.01],且相比于较低质量浓度白藜芦醇组(1μg/mL组),较高质量浓度白藜芦醇组(10μg/mL)更能诱导miR-146a的表达、抑制TNF-α蛋白的分泌(P＜0.01).结论白藜芦醇可以上调肺泡巨噬细胞内miR-146a的表达水平,推测miR-146a参与了白藜芦醇的抗炎过程.
목적 관찰백려호순대지다당(LPS)유도폐포거서세포미소RNA-146a(miR-146a)표체적영향.방법 장체외배양적대서폐포거서세포(NR8383)안2×106개/공접충우륙공판,세포첩벽90 min후,분위린산염완충액(PBS)대조조、종질량농도LPS(1μg/mL)처리조、백려호순(1 μg/mL)예처리30 min+종질량농도LPS(1μg/mL)처리조、백려호순(1 0μg/mL)예처리30 min+종질량농도LPS(1μg/mL)처리조.세포배양6h후,수집세포화세포배양상청액,채용실시형광정량PCR검측세포중miR-146a적표체변화,채용매련면역흡부실험(ELISA)검측세포배양상청액중TNF-α단백적표체변화.결과 여PBS대조조상비,LPS처리조세포중miR-146a화TNF-α단백표체량균명현승고[miR-146a표체배수(5.92±1.57)배vs.(1.03±0.58)배,P＜0.01;TNF-α질량농도(644.2±36.82) pg/mL vs.(26.15±13.43)pg/mL,P＜0.01].여LPS처리조상비,백려호순예처리조(1μg/mL화10 μg/mL)세포중miR-146a표체량균명현승고[miR-146a표체배수(8.67±1.18)배、(11.17±1.40)배vs.(5.92±1.57)배,P＜0.01],TNF-α단백질량농도균현저강저[TNF-α질량농도(447.7±41.48) pg/mL、(345.0±42.54)pg/mL vs.(644.2±36.82)pg/mL,P＜0.01],차상비우교저질량농도백려호순조(1μg/mL조),교고질량농도백려호순조(10μg/mL)경능유도miR-146a적표체、억제TNF-α단백적분비(P＜0.01).결론 백려호순가이상조폐포거서세포내miR-146a적표체수평,추측miR-146a삼여료백려호순적항염과정.
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.