Objective Puerarin can remove inflammatory mediators, but the underlying mechanisms are not yet clear.The purpose of this study is to observe the synthesis of IL-6 in co-cultured bronchial epithelial (Beas-2B) and neutrophil cells, and to investigate the inhibitory effect of puerarin on IL-6.Methods Beas-2B and neutrophil cells were extracted and divided into six groups: neutrophil cells cultured alone (NC), Beas-2B cells cultured alone (BC), Beas-2B and neutrophil cells co-cultured (BC+NC), and Beas-2B and neutrophil cells co-cultured with puerarin at 50 μg/mL (BC+NC+P50), 100 μg/mL (BC+NC+P100) and 200 μg/mL (BC+NC+P200).The concentrations of IL-6 in different groups were quantitatively measured by ELISA and the expression of the IL-6 gene detected by real-time PCR.Results After 18 hours of culturing, the concentration of IL-6 was significantly increased in the BC+NC group ([280.409±21.340] pg/mL) as compared with those in BC ([240.002±23.727] pg/mL), NC ([22.771±1.146] pg/mL), BC+NC+P50 ([244.205±9.335] pg/mL), BC+NC+P100 ([218.168±18.635] pg/mL), and BC+NC+P200 ([179.539±7.340] pg/mL) (P<0.05), but lower in the BC+NC+P200 than in the BC+NC+P50 and BC+NC+P100 groups (P<0.05).The expression of the IL-6 gene was markedly up-regulated in the BC+NC group (1.515±0.013) in comparison with those in BC (1.000±0.215), BC+NC+P50 (0.398±0.024), BC+NC+P100 (0.332±0.016), and BC+NC+P200 (0.306±0.015) (P<0.05), but lower in the BC+NC+P200 than in the BC and BC+NC+P50 groups (P<0.05).Conclusion Direct-contact co-culture of Beas-2B and neutrophil cells significantly enhances the synthesis and release of the inflammatory mediator IL-6, which can be inhibited by puerarin.%目的 葛根素可清除炎性介质,但具体机制尚不明确.文中旨在观察支气管上皮细胞(Beas-2B细胞)和嗜中性粒细胞(Neutrophils细胞)接触共培养体系中白细胞介素-6(IL-6)合成,探讨不同剂量葛根素对共培养体系IL-6的抑制作用.方法 提取支气管上皮细胞(Beas-2B细胞)和嗜中性粒细胞(Neutrophils细胞),并分为6组:粒细胞组(Neutrophils细胞单独培养)、上皮细胞组(Beas-2B细胞单独培养)、共培养组(Beas-2B+Neutrophils细胞接触共培养)以及共培养+葛根素200组、共培养+葛根素100组、共培养+葛根素50组(Beas-2B+Neutrophils+葛根素,调节葛根素终浓度分别为200、100、50μg/mL).接种密度为1×106/孔,进行上清液IL-6浓度分析.将各联合培养组中的Beas-2B和Neutrophils细胞分别分离提取,进行不同细胞IL-6基因表达分析. 结果 共培养18h,共培养组IL-6浓度[(280.409±21.340)pg/mL]较上皮细胞组[(240.002±23.727)pg/mL]、粒细胞组[(2.771±1.146)pg/mL]、共培养+葛根素50组[(244.205±9.335)pg/mL]、共培养+葛根素100组[(218.168±18.635)pg/mL]、共培养+葛根素200组[(179.539±7.340)pg/mL]明显增加(P<0.05);共培养+葛根素200组IL-6浓度较共培养+葛根素50组、共培养+葛根素100组明显降低(P<0.05).共培养组Beas-2B细胞IL-6基因表达(1.151±0.013)较上皮细胞组(1.000±0.215)、共培养+葛根素50组(0.398±0.024)、共培养+葛根素100组(0.332±0.016)、 共培养+葛根素200组(0.306±0.015)明显升高(P<0.05);共培养+葛根素200组Beas-2B细胞IL-6基因表达较上皮细胞组、共培养+葛根素50组明显降低(P<0.05). 结论 支气管上皮细胞和嗜中性粒细胞接触共培养可以促进炎性介质IL-6的合成与释放,并随着葛根素浓度升高,其合成的抑制逐渐增强.
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