【摘要】 目的·探讨3,3’-二吲哚基甲烷(3,3’-diindolylmethane,DIM)对脂多糖(lipopolysaccharide,LPS)诱导的人牙周膜细胞(human periodontal ligament cells,hPDLCs)分泌炎症因子的影响,并初步探讨其相关作用机制。方法·分离培养hPDLCs,CCK-8法测定DIM对hPDLCs增殖的影响,检测DIM毒性浓度范围。将hPDLCs分为4组:空白组加入不含LPS和DIM的无血清DMEM;LPS组仅加入含LPS(终浓度为10μg/mL)的无血清DMEM;低浓度组含10μg/mL LPS+6.25μmol/L DIM;高浓度组含10μg/mL LPS+12.50μmol/L DIM。培养12 h,酶联免疫吸附试验检测各组上清液中TNF-α、IL-1β和IL-6浓度,Western blotting检测hPDLCs内丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)和核因子κB(nuclear factorκB,NF-κB)信号通路中蛋白的变化。结果·DIM在50μmol/L范围内时细胞活力不受影响(P>0.05)。低浓度和高浓度DIM均抑制LPS刺激下hPDLCs分泌炎症因子TNF-α、IL-1β和IL-6(P<0.05),抑制效果随DIM的浓度升高而增强。DIM可显著抑制LPS激活NF-κB信号通路。结论·DIM可能通过抑制NF-κB信号通路的激活,抑制LPS诱导的hPDLCs分泌促炎因子TNF-α、IL-1β和IL-6。更多还原

【Abstract】 Objective·To investigate the effect of 3,3’-diindolylmethane(DIM) on the expression of inflammatory cytokines in human periodontal ligament cells(hPDLCs) induced by lipopolysaccharide(LPS) and to study the related mechanism. Methods·hPDLCs were isolated and cultured, and CCK-8 method was used to detect the effect of DIM on the proliferation of hPDLCs. hPDLCs were randomly divided into 4 groups: blank group(without LPS and DIM), LPS group(10 μg/mL LPS), 10 μg/mL LPS+6.25 μg/mL DIM, 10 μg/mL LPS+12.50 μg/mL DIM. The cells of all groups were cultured for 12 h. The protein levels of TNF-α, IL-1β and IL-6 in supernatant were detected by enzyme linked immunosorbent assay. The change of mitogenactivated protein kinase(MAPK) and nuclear factor κB(NF-κB) signaling pathways were detected by Western blotting. Results·The cell viability was not affected when the DIM concentration was less than 50 μmol/L(P>0.05). DIM at 6.25 and 12.50 μg/mL reduced the LPS-induced expression of TNF-α, IL-1β and IL-6 at protein levels(P<0.05). DIM inhibited the activation of the NF-κB signaling pathway. Conclusion·DIM can reduce the LPS-induced inflammatory cytokine expression in hPDLCs via restraining the activation of the NF-κB signaling pathway.更多还原