低水平激光照射对脂多糖介导的人牙周膜成纤维细胞炎症损伤的保护作用。
Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts.
机构信息
Dept. of Stomatology, Tianjin Jinnan Hospital, Tianjin 300350, China.
Dept. of Stomatology, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300192, China.
出版信息
Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Jun 1;39(3):260-266. doi: 10.7518/hxkq.2021.03.003.
OBJECTIVES
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 LPS. The three subgroups of the LPS+LLLI group were exposed to different LLLI. After 4 days, the cell apoptosis, viability, and intracellular free Ca 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 Caconcentration and decreased cell viability (<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels were higher in the cell supernatant (<0.05), and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs was significantly increased (<0.05). Compared with the LPS group, the LPS+LLLI group showed significantly decreased apoptosis rate and intracellular free Ca concentration and significantly increased cell viability (<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 (<0.05).
CONCLUSIONS
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.
目的
研究低水平激光照射（LLLI）对脂多糖（LPS）诱导的人牙周膜成纤维细胞（hPDLFs）炎性损伤的作用及机制。
方法
将 hPDLF 接种于培养板中，随机分为正常组、LPS 组和 LPS+LLLI 组。正常组细胞在常规培养基中培养，LPS 组和 LPS+LLLI 组细胞在含 1mg·L LPS 的 RPMI1640 培养基中培养。将 LPS+LLLI 组的三个亚组分别暴露于不同的 LLLI。4 天后，测量各组细胞凋亡率、细胞活力和细胞内游离 Ca 浓度。采用酶联免疫吸附试验（ELISA）检测各组肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-8、IL-1β 和 IL-6 的含量。采用逆转录-聚合酶链反应（RT-PCR）和 Western blot 检测各组 hPDLFs 基质金属蛋白酶（MMP）-2、MMP-3 和 MMP-9 基因和蛋白的表达。
结果
与正常组相比，LPS 组 hPDLFs 凋亡率升高，细胞内游离 Ca 浓度升高，细胞活力降低（P<0.05）。细胞上清液中 TNF-α、IL-8、IL-1β 和 IL-6 水平升高（P<0.05），hPDLFs MMP-2、MMP-3 和 MMP-9 基因和蛋白表达显著增加（P<0.05）。与 LPS 组相比，LPS+LLLI 组 hPDLFs 凋亡率和细胞内游离 Ca 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 和 IL-6 水平及 hPDLFs MMP-2、MMP-3 和 MMP-9 基因和蛋白表达均显著降低（P<0.05）。
结论
LLLI 对 LPS 诱导的 hPDLFs 炎性损伤具有保护作用，当辐照强度为 4 J·cm 时，效果最明显。
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