PMID- 34275223 OWN - NLM STAT- MEDLINE DCOM- 20210720 LR - 20210720 IS - 1002-0098 (Print) IS - 1002-0098 (Linking) VI - 56 IP - 7 DP - 2021 Jul 9 TI - [Role and mechanism of low-dose lipopolysaccharide-treated human periodontal ligament stem cells on the expression of macrophage pro-inflammatory factors]. PG - 672-678 LID - 10.3760/cma.j.cn112144-20210329-00146 [doi] AB - Objective: To investigate the effect of low dose lipopolysaccharide (LPS)-treated human periodontal ligament stem cells (HPDLSC) on the expression of macrophage pro-inflammatory factors and the mechanism involved. Methods: The primary HPDLSCs were obtained from healthy third molar periodontal ligament tissue. Phosphate buffer saline (PBS), 100 mug/L or 10 mg/L of LPS were used to treat HPDLSCs for 48 h, and their conditioned media were respectively co-cultured with THP-1-derived macrophages for 48 h. The corresponding experimental groups were PBS-treated HPDLSC-derived conditioned medium (CM-C) group, low dose LPS-treated HPDLSC-derived conditioned medium (CM-L) group, and high dose LPS-treated HPDLSC-derived conditioned medium (CM-H) group. Quantitative real-time PCR was performed to explore the mRNA expressions of macrophage interleukin (IL)-6, IL-8, IL-12, tumor necrosis factor-alpha (TNF-alpha) in the CM-C, CM-L and CM-H groups, and the expressions of nuclear factor (erythroid-derived 2)-like 2 (NRF2), glutamate-cysteine ligase catalytic subunit (GCLC), NAD(P)H quinone dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) in the CM-C and CM-L groups. Meanwhile, Western blotting was used to detect the change of nuclear and cytoplasmic NRF2 and the levels of GCLC and HO-1 in the CM-C and CM-L groups. The 2', 7'-dichlorofluorescein probe was adopted to detect the reactive oxygen species (ROS) levels of macrophages in the CM-C and CM-L groups and the data were characterized by the mean fluorescent intensity (MFI). Results: The mRNA expressions of macrophage pro-inflammatory factors IL-6, IL-8, IL-12 and TNF-alpha in the CM-H group (2.332+/-0.594, 3.601+/-0.639, 2.120+/-0.677 and 2.468+/-0.236) were significantly upregulated compared with those in the CM-C group (1.000+/-0.321, 1.000+/-0.151, 1.000+/-0.059 and 1.000+/-0.095) (P<0.05); while the relative mRNA levels of IL-6, IL-12 and TNF-alpha in the CM-L group (0.056+/-0.002, 0.215+/-0.024 and 0.567+/-0.071) were much lower than those in the CM-C group (1.000+/-0.209, 1.000+/-0.220 and 1.000+/-0.220) (P<0.05). At the mRNA level, the expression of NRF2 was significantly increased in the CM-L group (1.864+/-0.198) compared with that in the CM-C group (1.000+/-0.094) (P<0.05). At the protein level, the cytoplasmic NRF2 and nuclear NRF2 were increased in CM-L group (1.175+/-0.104 and 1.308+/-0.082) compared with those in the CM-C group (1.000+/-0.025 and 1.000+/-0.049) (P<0.05). Furthermore, the antioxidative genes, i.e. GCLC and NQO1, localized in NRF2 downstream, were significantly upregulated in the CM-L group (1.786+/-0.278 and 1.444+/-0.078) compared with the CM-C group (1.000+/-0.139 and 1.000+/-0.226) (P<0.05). The protein levels of GCLC and HO-1 were augmented in the CM-L group (1.159+/-0.036 and 1.412+/-0.075) in contrast with those in the CM-C group (1.000+/-0.050 and 1.000+/-0.013) (P<0.05). In addition, the MFI in the CM-L group (123 419+/-1 302) was significantly lower than that in the CM-C group (139 193+/-1 241) (P<0.05). Conclusions: Low-dose LPS-treated HPDLSCs could regulate oxidative stress response through activating the NRF2 signaling pathway of macrophages and further downregulating the expressions of macrophage pro-inflammatory factors. FAU - Wang, Y Z AU - Wang YZ AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. FAU - Fei, D D AU - Fei DD AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. FAU - Zhang, Y AU - Zhang Y AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. FAU - Zhang, X G AU - Zhang XG AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. FAU - Wang, Y AU - Wang Y AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. FAU - Wang, Q T AU - Wang QT AD - Department of Periodontology, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Xi'an 710032, China. LA - chi GR - 81771069/National Natural Science Foundation of China/ PT - Journal Article PL - China TA - Zhonghua Kou Qiang Yi Xue Za Zhi JT - Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology JID - 8711066 RN - 0 (Lipopolysaccharides) RN - 0 (NF-E2-Related Factor 2) RN - EC 1.14.14.18 (Heme Oxygenase-1) SB - IM MH - Heme Oxygenase-1/genetics MH - Humans MH - *Lipopolysaccharides MH - Macrophages MH - NF-E2-Related Factor 2 MH - *Periodontal Ligament MH - Stem Cells EDAT- 2021/07/19 06:00 MHDA- 2021/07/21 06:00 CRDT- 2021/07/18 22:22 PHST- 2021/07/18 22:22 [entrez] PHST- 2021/07/19 06:00 [pubmed] PHST- 2021/07/21 06:00 [medline] AID - 10.3760/cma.j.cn112144-20210329-00146 [doi] PST - ppublish SO - Zhonghua Kou Qiang Yi Xue Za Zhi. 2021 Jul 9;56(7):672-678. doi: 10.3760/cma.j.cn112144-20210329-00146.