低能量激光对人牙周膜细胞白细胞介素-6、肿瘤坏死因子-α、骨保护素、核因子-κB受体活化因子配体表达的影响  

Effects of low-level laser on the expression of interleukin-6,tumor necrosis factor‑α,osteoprotegerin,and receptor activator of nuclear factor-κB ligand in human periodontal ligament cells

作　　者：汤盟 崔占琴[1] 王阳阳[1] 陈增国 李文静[2] 张翠萍 Tang Meng;Cui Zhanqin;Wang Yangyang;Chen Zengguo;Li Wenjing;Zhang Cuiping(Dept.of Orthodontics,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Dept.of Oral Medicine,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Dept.of Stomatology,Yongding District Hospital of Longyan City,Fujian Province,Longyan 364100,China)

机构地区：[1]河北医科大学第二医院口腔正畸科,石家庄050000 [2]河北医科大学第二医院口腔内科,石家庄050000 [3]福建省龙岩市永定区医院口腔科,龙岩364100

出　　处：《华西口腔医学杂志》2023年第5期521-532,共12页West China Journal of Stomatology

基　　金：2022年河北省政府资助临床医学优秀人才项目[冀财预复(2022)180号]。

摘　　要：目的通过观察高糖环境下低能量激光(LLL)对受静压力刺激的人牙周膜细胞(HPDLC)白细胞介素-6(IL-6)、肿瘤坏死因子(TNF)-α、骨保护素(OPG)、核因子-κB受体活化因子配体(RANKL)表达的影响,以期探究LLL对糖尿病患者牙周组织炎症及正畸治疗过程中骨改建调控的分子生物学机制。方法体外培养HPDLC,模拟正畸加力,结合LLL照射,将所培养细胞随机分为4组:低糖型杜氏改良Eagle培养基(DMEM)+压力刺激(A组),高糖型DMEM+压力刺激(B组),低糖型DMEM+LLL照射+压力刺激(C组),高糖型DMEM+LLL照射+压力刺激(D组),其中C、D组根据给予的激光能量密度值不同又分C1、D1组(能量密度值:3.75 J/cm2)和C2、D2组(能量密度值:5.625 J/cm2)。根据已有分组,对A、B、C、D组细胞进行加力,对C、D组细胞在细胞加力前进行LLL照射。分别观察0、12、24、48、72 h,定时提取各组细胞培养上清液,采用酶联免疫吸附(ELISA)法检测各组不同时段IL-6、TNF-α、OPG、RANKL的蛋白表达。结果1)在持续静压力刺激下,HPDLC分泌的IL-6、TNF-α浓度随时间逐渐上升;12 h后IL-6、TNF-α的浓度在A组与B、C1、C2组组间两两比较的差异均有统计学意义(P<0.05),B组与D1、D2组组间两两比较的差异也均有统计学意义(P<0.01)。2)OPG蛋白浓度在24h之前呈现出上升趋势,24h后随时间出现下降趋势;RANKL蛋白浓度随时间呈上升趋势;OPG/RANKL比值随时间呈下降趋势;持续静压力刺激12h后,A组与B、C1、C2组,B组与D1、D2组组间两两比较的OPG、RANKL及OPG/RANKL的比值的差异均具有统计学意义(P<0.05)。结论1)在高糖环境持续静压力刺激下,HPDLC分泌的IL-6、TNF-α浓度随时间呈现上升趋势;OPG的表达水平降低,RANKL的表达水平增加,OPG/RANKL的比值减小,提示高糖环境会促进骨吸收反应的发生;给予LLL照射干预后发现,IL-6、TNF-α的浓度出现下降,表明其可拮抗高糖环境所致的炎症因子水平的升高,并�Objective This study aims to determine the effects of low-level laser(LLL)on the expression of interleukin-6(IL-6),tumor necrosis factor(TNF)-α,osteoprotegerin(OPG),and receptor activator of nuclear factor-κB ligand(RANKL)in human periodontal ligament cells(HPDLCs)stimulated by high glucose;and identify the molecular mechanism of LLL therapy in the regulation of periodontal inflammation and bone remodeling during orthodontic treatment in diabetic patients.Methods HPDLCs were cultured in vitro to simulate orthodontic after loading and irradiated with LLL therapy.The cultured cells were randomly divided into four groups:low glucose Dulbecco’s modification of Eagle’s medium(DMEM)+stress stimulation(group A),high glucose DMEM+stress stimulation(group B),hypoglycemic DMEM+LLL therapy+stress stimulation(group C),and hyperglycemic DMEM+LLL therapy+stress stimulation(group D).Groups C and D were further divided into C1 and D1(energy density:3.75 J/cm2)and C2 and D2(energy density:5.625 J/cm2).Cells in groups A,B,C,and D were irradiated by LLL before irradiation.At 0,12,24,48,and 72 h,the supernatants of the cell cultures were extracted at regular intervals,and the protein expression levels of IL-6,TNF-α,OPG,and RANKL were detected by enzyme-linked immunosorbent assay.Results 1)The levels of IL-6 and TNF-αsecreted by HPDLCs increased gradually with time under static pressure stimulation.After 12 h,the levels of IL-6 and TNF-αsecreted by HPDLCs in group A were significantly higher than those in groups B,C1,and C2(P<0.05),which in group B were significantly higher than those in groups D1,and D2(P<0.01).2)The OPG protein concentration showed an upward trend before 24 h and a downward trend thereafter.The RANKL protein concentration increased,whereas the OPG/RANKL ratio decreased with time.Significant differences in OPG,RANKL,and OPG/RANKL ratio were found among group A and groups B,C1,C2 as well as group B and groups D1,D2(P<0.05).Conclusion 1)In the high glucose+stress stimulation environment,the concentrations of

关 键 词：高糖环境 白细胞介素-6 肿瘤坏死因子-Α 骨保护素 核因子-ΚB受体活化因子配体 人牙周膜细胞 低能量激光 

分 类 号：R783.5[医药卫生—口腔医学]