Effect of MicroRNA-218-5p on Inflammation of Human Periodontal Ligament Stem Cells Induced by Lipopolysaccharide through Targeting TLR2 Gene

doi:10.13701/j.cnki.kqyxyj.2020.12.007

Abstract

Abstract: Objective: To investigate the effects and mechanism of miR-218-5p on proliferation, apoptosis, and inflammation of periodontal ligament stem cells (PDLSCs) induced by lipopolysaccharide (LPS). Methods: PDLSCs were isolated and cultured. PDLSCs cells were treated with LPS to establish an inflammatory model. MTT assay and flow cytometry were applied to detect the cell survival rate and apoptosis rate, respectively. qRT-PCR and Western blot were performed to detect the expression levels of miR-218-5p and TLR2 protein, respectively. The kits was used to detect the contents of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in PDLSCs. The dual luciferase reporting system was used to detect the targeting relationship between miR-218-5p and TLR2. Results: Human PDLSs were treated by LPS for 0h, 12h, 24h, and 48h. miR-218-5p in PDLSs cells was down-regulated, TLR2 was up-regulated, cell survival rate was down-regulated, apoptosis rate was up-regulated, and the contents of inflammatory factors IL-6, IL-1β, and TNF-α were up-regulated, showing a time-dependent trend with statistical significance (P<0.05). MiR-218-5p targeted and regulated the expression of TLR2. Over-expression of miR-218-5p or silencing of TLR2 both improved cell survival rate and reduced cell apoptosis rate and levels of inflammatory factors IL-6, IL-1β, and TNF-α. Over-expression of TLR2 reversed the effect of miR-218-5p over-expression on inflammation and proliferation of PDLSCs induced by LPS. Conclusion: MiR-218-5p regulates the proliferation, apoptosis, and inflammatory response of PDLSCs cells induced by LPS through targeting TLR2.

Key words: microRNA-218-5p, lipopolysaccharide, Toll-like receptor 2, periodontal membrane stem cells, inflammation, survival rate, apoptosis

ZHOU Yang. Effect of MicroRNA-218-5p on Inflammation of Human Periodontal Ligament Stem Cells Induced by Lipopolysaccharide through Targeting TLR2 Gene[J]. Journal of Oral Science Research, 2020, 36(12): 1108-1112.

References

[1] 张玉杰.牙周病研究进展[J].现代中西医结合杂志,2015,24(27):3071-3073.
[2] 袁萍,李淑慧,赵璐,等.炎症微环境下人牙周膜干细胞的生物学特性[J].中国组织工程研究,2016,20(6):898-905.
[3] 孔晨,李永丽,李思佳,等.牙周炎动物模型的研究进展[J].医学综述,2019,25(17):3344-3349.
[4] 王宝华,赵亦非,沈莉蓉,等.牙周炎组织中miRNA表达谱差异筛选及功能预测分析[J].上海口腔医学, 2019, 28(4):408-411.
[5] Zhang T, Xiang L. Honokiol alleviates sepsis-induced acute kidney injury in mice by targeting the miR-218-5p/heme oxygenase-1 signaling pathway [J]. Cell Mol Biol Lett, 2019, 24:15.
[6] 郑富军,马芬,田小龙.慢性牙周炎患者牙龈组织中TLR2、TLR4、CD14的表达水平及意义[J].临床和实验医学杂志,2018,17(19):67-70.
[7] 冯娜,李珂,邵江红.miR-146a在人炎症牙周膜干细胞中的表达及其与TLR4靶向调控作用的研究[J].牙体牙髓牙周病学杂志,2018,28(1):1-5.
[8] 刘大勇,周伟伟,高润涛.TNF-α及LPS刺激下牙周膜干细胞IkB磷酸化及其下游基因的表达[J].牙体牙髓牙周病学杂志,2013,23(8):523-527.
[9] Amaral SA, Pereira TSF, Brito JAR, et al. Comparison of miRNA expression profiles in individuals with chronic or aggressive periodontitis [J]. Oral Dis, 2019, 25(2):561-568.
[10] Yu SF, Feng WY, Chai SQ, et al. Down-regulation of miR-218-5p promotes apoptosis of human umbilical vein endothelial cells through regulating high-mobility group box-1 in henoch-schonlein purpura [J]. Am J Med Sci, 2018, 356(1):64-71.
[11] Wu Z, Han Y, Li Y, et al. MiR-218-5p inhibits the stem cell properties and invasive ability of the A2B5⁺ CD133^- subgroup of human glioma stem cells [J]. Oncol Rep, 2016, 35(2):869-877.
[12] Song J, Wang QH, Zou SC. Role of microRNA-218-5p in the pathogenesis of chronic obstructive pulmonary disease [J]. Eur Rev Med Pharmacol Sci, 2018, 22(13):4319-4324.
[13] Guo J, Zeng X, Miao J, et al. MiRNA-218 regulates osteoclast differentiation and inflammation response in periodontitis rats through Mmp9 [J]. Cell Microbiol, 2019, 21(4):e12979.
[14] 熊正方,李冰,党小红.Toll样受体2功能的研究进展[J].中华生物医学工程杂志, 2014, 20(1):71-74.
[15] 龙晏,邱申彩,李淑慧,等.过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响[J].口腔医学研究,2019,35(1):28-32.
[16] Blufstein A, Behm C, Gahn J, et al. Synergistic effects triggered by simultaneous Toll-like receptor-2 and -3 activation in human periodontal ligament stem cells[J]. J Periodontol, 2019, 90(10):1190-1201.