骨化三醇对牙龈卟啉单胞菌脂多糖诱导的人牙周膜细胞IL-8、IL-6表达的影响研究  

Effect of calcitriol on IL-8 and IL-6 expression in Porphyromonas gingivalis lipopolysaccharide-induced human periodontal ligament cells

作　　者：唐晓琳[1] 刘俊超[1] 包穆蓉[2] 孙尚敏[1] TANG Xiao-lin LIU Jun-chao BAO Mu-rong SUN Shang-min(Department of Periodontology, School and Hospital of Stomatology, China Medical University, Shettyang 110002, China)

机构地区：[1]中国医科大学口腔医学院.附属口腔医院牙周病科,辽宁沈阳110002 [2]中国医科大学口腔医学院.附属口腔医院牙体牙髓病科,辽宁沈阳110002

出　　处：《中国实用口腔科杂志》2016年第12期734-737,742,共5页Chinese Journal of Practical Stomatology

基　　金：辽宁省自然科学基金项目计划(2014021073);辽宁省科学技术计划(2013225303)

摘　　要：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 explore the effect of ealcitriol on IL-8 and IL-6 expression in Porphyromonas gingivalis lipo- polysaccharide （Pg-LPS）-induced human periodontal ligament cells. Methods Primary cultures of hPDLCs were es- tablished and obtained from the extracted premolars of 3 patients for orthodontic treatment, hPDLCs of were respectively treated with 0.1% absolute ethyl alcohol （control group）, 10 μg/mL Pg-LPS （simple Pg-LPS group, A）, 10^-10mol/L 1, 25D （low 1.25 D group,B）, 10^-8 mol/L 1,25D （High 1.25 D group,C）, 10 ^-10mol/L 1,25D＋10 μg/mL Pg-LPS （group D） , 10^-8mol/L 1,25D＋ 10 μg/mL Pg-LPS （group E） for 24 h and 48 h. The supernatant fluid samples were collected and the IL-8 and IL-6 concentration was determined with enzyme-linked immunosorbent assay （ELISA） method. Results （ 1 ） The 10 μg/mL Pg-LPS exerted the highest promotion effcot on the expression level of IL-8 by 38.86-folds at 48 h （P 〈 0.001 ） and that of IL-6 by 6.19-folds at 24 h （P 〈 0.001 ） respectively. （2） 1, 25D down-regulated the IL-8 level in a dose- and timedependent manner. The highest inhibition effect occurred at 48 h, the IL-8 level of cells treat-ed with 10^-8moL/L l, 25D was 49.94% of that of the control cells （P 〈 0.001）. （3） 1,25D significantly decreased the LPS-indueed IL-8 and IL-6 production. At 48 h, 10^-8μmol/L 1, 25D combined with Pg-LPS significantly down-regulated the IL-8 level in hPDLCs, 71.98% of that in the cells treatedwith Pg-LPS alone（P 〈 0.001）. （4） At 24 h, the level of IL-6 in cells treated with Pg-LPS combined with 10^-8mol/L 1, 25D was 84.51% of that in ceils treated with Pg-LPS alone （P=0.003）. Conclusion Our results suggest that vitamin D may suppress the periodontal inflammation partly by inhibiting the expressions of innate IL-8 and Pg-LPS-induced IL-8 and IL-6 in hPDLCs.

关 键 词：牙周膜细胞 维生素D 牙龈卟啉单胞菌 脂多糖 白细胞介素-6 白细胞介素-8 

分 类 号：R78[医药卫生—口腔医学]