华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (3): 260-266.doi: 10.7518/hxkq.2021.03.003

• 基础研究 • 上一篇    

低能量激光照射对脂多糖介导人牙周膜成纤维细胞炎性损伤的保护作用

靳晓兰1(), 张亚男2, 孙成蕊1, 邹朝晖3()   

  1. 1.天津市津南医院口腔科,天津 300350
    2.天津中医药大学第一附属医院口腔科,天津 300192
    3.天津医科大学口腔医院牙体牙髓科三室,天津 300070
  • 收稿日期:2020-04-09 修回日期:2021-03-07 出版日期:2021-06-01 发布日期:2021-05-26
  • 通讯作者: 邹朝晖 E-mail:jinxiaolan2014@sina.com;zouzhaohui2005@sina.com
  • 作者简介:靳晓兰,主治医师,硕士,E-mail:jinxiaolan2014@sina.com

Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts

Jin Xiaolan1(), Zhang Yanan2, Sun Chengrui1, Zou Zhaohui3()   

  1. 1.Dept. of Stomatology, Tianjin Jinnan Hospital, Tianjin 300350, China
    2.Dept. of Stomatology, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300192, China
    3.Dept. of Endodontics Third Room, School of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • Received:2020-04-09 Revised:2021-03-07 Online:2021-06-01 Published:2021-05-26
  • Contact: Zou Zhaohui E-mail:jinxiaolan2014@sina.com;zouzhaohui2005@sina.com

摘要: 目的

研究低能量激光照射(LLLI)对脂多糖(LPS)介导人牙周膜成纤维细胞(hPDLFs)炎性损伤的影响及机制。

方法

将hPDLFs接种于培养板内,随机分为正常组、LPS组和LPS+LLLI组。正常组细胞行常规培养基培养,LPS组及LPS+LLLI组在加入1 mg·L-1 LPS培养基中培养24 h,LPS+LLLI组3个亚组分别接受不同强度照射。4 d后检测各组细胞凋亡、活力及细胞内游离Ca2+浓度,酶联免疫吸附试验检测各组细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-8、IL-1β及IL-6的含量,逆转录聚合酶链反应(RT-PCR)及Western blot检测各个实验组hPDLFs的基质金属蛋白酶(MMP)-2、MMP-3、MMP-9基因和蛋白的表达。

结果

与正常组比较,LPS组hPDLFs细胞凋亡率及细胞内游离Ca2+浓度增加,细胞活力降低(P<0.05),细胞上清液中TNF-α、IL-8、IL-1β及IL-6的含量显著升高(P<0.05),hPDLFs的MMP-2、MMP-3、MMP-9基因和蛋白的表达显著升高(P<0.05)。与LPS组比较,LPS+LLLI组细胞凋亡率及细胞内游离Ca2+浓度降低,细胞活力升高(P<0.05),细胞上清液中TNF-α、IL-8、IL-1β及IL-6的含量显著降低(P<0.05),hPDLFs的MMP-2、MMP-3、MMP-9基因和蛋白的表达显著降低(P<0.05)。

结论

LLLI对LPS诱导 hPDLFs炎性损伤有保护作用,照射强度为4 J·cm-2时效果最明显。

关键词: 低能量激光照射, 人牙周膜成纤维细胞, 脂多糖, 炎性损伤, 影响

Abstract: Objective

To study the effect and mechanism of low-level laser irradiation (LLLI) on lipopolysaccharide (LPS)-induced inflammatory injury of human periodontal ligament fibroblasts (hPDLFs).

Methods

hPDLFs were inoculated into well plates and randomly divided into the normal group, LPS group, and LPS+LLLI group. The cells in the normal group were cultured in conventional medium. The hPDLFs in the LPS and LPS+LLLI groups were cultured in RPMI1640 medium containing 1 mg·L-1 LPS. The three subgroups of the LPS+LLLI group were exposed to different LLLI. After 4 days, the cell apoptosis, viability, and intracellular free Ca2+ concentration of each group were measured. The contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-8, IL-1β, and IL-6 were measured by enzyme linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of matrix metalloproteinase (MMP)-2, MMP-3, and MMP-9 genes and proteins of hPDLFs in each group.

Results

Compared with the normal group, the LPS group showed increased apoptosis rate of hPDLFs and intracellular free Ca2+concentration and decreased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels were higher in the cell supernatant (P<0.05), and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs was significantly increased (P<0.05). Compared with the LPS group, the LPS+LLLI group showed significantly decreased apoptosis rate and intracellular free Ca2+ concentration and significantly increased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels in the supernatant of cells and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs were significantly decreased (P<0.05).

Conclusion

LLLI has a protective effect on the inflammatory injury of hPDLFs induced by LPS, and the effect is most obvious when the irradiation intensity is 4 J·cm-2.

Key words: low-level laser irradiation, human periodontal ligament fibroblasts, lipopolysaccharide, inflammatory injury, effect

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