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目的观察活性形式的维生素D——骨化三醇(1,25D)对牙龈卟啉单胞菌脂多糖(Pg-LPS)诱导的人牙周膜细胞(h PDLCs)白细胞介素(IL)-8、IL-6表达的影响。方法取3例患者因正畸需要拔除的前磨牙牙周膜,组织块法原代培养h PDLCs,分别用0.1%无水乙醇(对照组)、10μg/m L Pg-LPS(单纯Pg-LPS组)、10~(-10)mol/L 1,25D(低浓度1,25D组)、10~(-8)mol/L 1,25D(高浓度1,25D组)、10~(-10)mol/L 1,25D+10μg/m L Pg-LPS(低浓度1,25D联合Pg-LPS组)、10~(-8)mol/L 1,25D+10μg/m L Pg-LPS(高浓度1,25D联合Pg-LPS组)处理第5代细胞。24和48 h后收集培养基上清液,酶联免疫吸附试验(ELISA)法检测h PDLCs IL-8和IL-6的表达水平。结果 (1)10μg/m L PgLPS处理h PDLCs 48 h时,其IL-8表达水平最高,为对照组的38.86倍(P<0.001);处理h PDLCs 24 h时,其IL-6表达水平最高,为对照组的6.19倍(P<0.001)。(2)1,25D以剂量和时间依赖方式抑制h PDLCs IL-8的内源性表达。10~(-8)mol/L 1,25D处理h PDLCs 48 h时,其IL-8表达水平为对照组的49.94%(P<0.001)。(3)1,25D显著抑制Pg-LPS诱导的h PDLCs IL-8和IL-6的表达。处理48 h时,高浓度1,25D联合Pg-LPS组h PDLCs IL-8的表达水平为单纯Pg-LPS组的71.98%(P<0.001);处理24 h时,高浓度1,25D联合Pg-LPS组h PDLCs IL-6的表达水平为单纯Pg-LPS组的84.51%(P=0.003)。结论 1,25D可以抑制h PDLCs IL-8的内源性表达,并抑制Pg-LPS诱导的h PDLCs IL-8和IL-6的表达,从而可能抑制牙周炎症反应。
Objective To observe the effects of active vitamin D (Ⅲ) calcitriol (1,25D) on interleukin-6 (IL-1β) in human PDLCs induced by Pg-LPS, 8, IL-6 expression. Methods The PD PDs were obtained from the periodontal ligament of the premolar to be removed by orthodontic treatment in three patients. Primary cultured PD PDs were treated with 0.1% absolute ethanol (control group), 10 μg / ml Pg-LPS 10 ~ (-10) mol / L 1,25D (low concentration 1,25D), 10 ~ (-8) mol / L 1,25D (high concentration 1,25D), 10 ~ (-10) (P <0.05), P (subscript a), P (subscript m) and Pg-LPS in low concentration 1,25D group and Pg-LPS group 5th and 5th generation cells were treated with 1, 25D and Pg-LPS groups. Supernatants were harvested after 24 and 48 h, and the levels of IL-8 and IL-6 in h PDLCs were detected by enzyme linked immunosorbent assay (ELISA). Results (1) The hPLCs treated with 10 μg / mL PgLPS had the highest level of IL-8 at 48 h, which was 38.86 folds higher than that of the control group (P <0.001). The h PDLCs had the highest IL-6 expression at 24 h , 6.19 times that of the control group (P <0.001). (2) 1,25D inhibited the endogenous expression of h-PDLCs IL-8 in a dose-and time-dependent manner. The expression of IL-8 in h PDLCs treated with 10 -8 mol / L 1,25 D for 48 h was 49.94% (P <0.001) of the control group. (3) 1,25D significantly inhibited Pg-LPS-induced IL-8 and IL-6 expression in h PDLCs. At 48 h, the expression levels of IL-8 in h PDLCs in high concentration 1,25 D group and Pg-LPS group were 71.98% (P <0.001) in Pg-LPS group alone. When treated with high concentrations of 1,25 D and Pg The expression level of IL-6 in PDLCs in LPS group was 84.51% (P = 0.003) in pure Pg-LPS group. Conclusion 1,25D can inhibit the endogenous expression of IL-8 in h PDLCs and inhibit the expression of IL-8 and IL-6 in h PDLCs induced by Pg-LPS, which may inhibit the periodontal inflammation.