高糖炎性环境下KLF4调控人牙周膜干细胞能量代谢的机制初探

Abstract

Abstract: Objective:To investigate the effects of Krüppel-like factor 4 (KLF4) in a high glucose and inflammatory environment on mitochondrial energy metabolism and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). Methods:The localization and expression of KLF4 in hPDLSCs was investigated under high glucose inflammatory environment using immunofluorescence and western blot. ALP staining and alizarin red staining were used to study osteogenic differentiation of hPDLSCs in hyperglycemic inflammatory environment. Lentivirus knockdown and overexpression techniques were used to detect the expression of KLF4 after virus infection by qRT-PCR and Western blot. CUT&RUN technique was used to analyze the changes of KLF4 binding DNA and related signaling pathways in hPDLSCs under high glucose inflammatory environment. The impact of KLF4 on oxidative respiratory capacity of hPDLSCs was explored using a Seahorse energy metabolism analyzer. Results:KLF4 expression was downregulated under high glucose inflammatory conditions, accompanied by reduced ALP activity, calcium nodule formation (all P<0.001), and expression of RUNX2 and OCN (P<0.001). Knockdown of KLF4 resulted in a significant decrease in ALP activity (P<0.001) and reduced calcium nodule formation (P<0.001) in hPDLSCs, while overexpression of KLF4 promote osteogenic differentiation. Under high glucose inflammatory environment, the downstream regulatory genes of KLF4 were enriched in energy metabolism, osteogenesis and other related pathways. KLF4 knockdown reduced OCR levels and increased ECAR levels in hPDLSCs, while KLF4 overexpression increased OCR levels and decreased ECAR levels. Conclusions:In a high glucose inflammatory environment, the osteogenic differentiation ability of hPDLSCs was compromised, and KLF4 may enhance the mitochondrial oxidative phosphorylation capacity to restore this ability.

Key words: human periodontal ligament stem cells, Krüppel-like factor 4, periodontitis, diabetes mellitus, mitochondrial energy metabolism

References 34

[1]	Genco R J, Borgnakke W S. Diabetes as a potential risk for periodontitis: association studies[J]. Periodontol 2000, 2020, 83(1): 40-45.
[2]	Sapey E, Yonel Z, Edgar R, et al. The clinical and inflammatory relationships between periodontitis and chronic obstructive pulmonary disease[J]. J Clin Periodontol, 2020, 47(9): 1040-1052.
[3]	Nibali L, Gkranias N, Mainas G, et al. Periodontitis and implant complications in diabetes[J]. Periodontol 2000, 2022, 90(1): 88-105.
[4]	Jepsen S, Caton J G, Albandar J M, et al. Periodontal manifestations of systemic diseases and developmental and acquired conditions: Consensus report of workgroup 3 of the 2017 World Workshop on the Classification of Periodontal and Peri‐Implant Diseases and Conditions[J]. Journal of clinical periodontology, 2018, 45: S219-S229.
[5]	Teeuw W J, Kosho M X, Poland D C, et al. Periodontitis as a possible early sign of diabetes mellitus[J]. BMJ Open Diabetes Res Care, 2017, 5(1): e000326.
[6]	Lalla E, Papapanou P N. Diabetes mellitus and periodontitis: a tale of two common interrelated diseases[J]. Nat Rev Endocrinol, 2011, 7(12): 738-48.
[7]	Liu J, Chen B, Bao J, et al. Macrophage polarization in periodontal ligament stem cells enhanced periodontal regeneration[J]. Stem Cell Res Ther, 2019, 10(1): 320.
[8]	Li W, Huang X, Yu W, et al. Activation of Functional Somatic Stem Cells Promotes Endogenous Tissue Regeneration[J]. J Dent Res, 2022, 101(7): 802-811.
[9]	Kato H, Taguchi Y, Tominaga K, et al. High Glucose Concentrations Suppress the Proliferation of Human Periodontal Ligament Stem Cells and Their Differentiation Into Osteoblasts[J]. J Periodontol, 2016, 87(4): e44-51.
[10]	Zheng J, Chen S, Albiero M L, et al. Diabetes Activates Periodontal Ligament Fibroblasts via NF-κB In Vivo[J]. J Dent Res, 2018, 97(5): 580-588.
[11]	Mosteiro L, Pantoja C, Alcazar N, et al. Tissue damage and senescence provide critical signals for cellular reprogramming in vivo[J]. Science, 2016, 354(6315): aaf4445.
[12]	Oka S I, Chin A, Park J Y, et al. Thioredoxin-1 maintains mitochondrial function via mechanistic target of rapamycin signalling in the heart[J]. Cardiovasc Res, 2020, 116(10): 1742-1755.
[13]	Wu C Z, Yuan Y H, Liu H H, et al. Epidemiologic relationship between periodontitis and type 2 diabetes mellitus[J]. BMC Oral Health, 2020, 20(1): 204.
[14]	Preshaw P M, Alba A L, Herrera D, et al. Periodontitis and diabetes: a two-way relationship[J]. Diabetologia, 2012, 55(1): 21-31.
[15]	Zhang R, Liang Q, Kang W, et al. Metformin facilitates the proliferation, migration, and osteogenic differentiation of periodontal ligament stem cells in vitro[J]. Cell Biol Int, 2020, 44(1): 70-79.
[16]	Jiao J, Jing W, Si Y, et al. The prevalence and severity of periodontal disease in Mainland China: Data from the Fourth National Oral Health Survey (2015-2016)[J]. J Clin Periodontol, 2021, 48(2): 168-179.
[17]	Kominato H, Takeda K, Mizutani K, et al. Metformin accelerates wound healing by Akt phosphorylation of gingival fibroblasts in insulin-resistant prediabetes mice[J]. J Periodontol, 2022, 93(2): 256-268.
[18]	Zhang D, Lin W, Jiang S, et al. Lepr-Expressing PDLSCs Contribute to Periodontal Homeostasis and Respond to Mechanical Force by Piezo1[J]. Adv Sci (Weinh), 2023, 10(29): e2303291.
[19]	Wang L, Liu C, Wu F. Low-level laser irradiation enhances the proliferation and osteogenic differentiation of PDLSCs via BMP signaling[J]. Lasers Med Sci, 2022, 37(2): 941-948.
[20]	Quan H, Dai X, Liu M, et al. Luteolin supports osteogenic differentiation of human periodontal ligament cells[J]. BMC Oral Health, 2019, 19(1): 229.
[21]	Luo Y, Gou H, Chen X, et al. Lactate inhibits osteogenic differentiation of human periodontal ligament stem cells via autophagy through the MCT1-mTOR signaling pathway[J]. Bone, 2022, 162: 116444.
[22]	Zhao P, Yue Z, Nie L, et al. Hyperglycaemia-associated macrophage pyroptosis accelerates periodontal inflamm-aging[J]. J Clin Periodontol, 2021, 48(10): 1379-1392.
[23]	Ebersole J L, Graves C L, Gonzalez O A, et al. Aging, inflammation, immunity and periodontal disease[J]. Periodontol 2000, 2016, 72(1): 54-75.
[24]	Chen Y C, Liao B K, Lu Y F, et al. Zebrafish Klf4 maintains the ionocyte progenitor population by regulating epidermal stem cell proliferation and lateral inhibition[J]. PLoS Genet, 2019, 15(4): e1008058.
[25]	Müller M, Schaefer M, F?h T, et al. Argonaute proteins regulate a specific network of genes through KLF4 in mouse embryonic stem cells[J]. Stem Cell Reports, 2022, 17(5): 1070-1080.
[26]	Konishi H, Rahmawati F N, Okamoto N, et al. Discovery of Transcription Factors Involved in the Maintenance of Resident Vascular Endothelial Stem Cell Properties[J]. Mol Cell Biol, 2024, 44(1): 17-26.
[27]	Katz J P, Perreault N, Goldstein B G, et al. Loss of Klf4 in mice causes altered proliferation and differentiation and precancerous changes in the adult stomach[J]. Gastroenterology, 2005, 128(4): 935-45.
[28]	Lu Y, Brommer B, Tian X, et al. Reprogramming to recover youthful epigenetic information and restore vision[J]. Nature, 2020, 588(7836): 124-129.
[29]	Blacher E, Tsai C, Litichevskiy L, et al. Aging disrupts circadian gene regulation and function in macrophages[J]. Nat Immunol, 2022, 23(2): 229-236.
[30]	Li Y, Xiong Z, Jiang Y, et al. Klf4 deficiency exacerbates myocardial ischemia/reperfusion injury in mice via enhancing ROCK1/DRP1 pathway-dependent mitochondrial fission[J]. J Mol Cell Cardiol, 2023, 174: 115-132.
[31]	Han Y, He M, Marin T, et al. Roles of KLF4 and AMPK in the inhibition of glycolysis by pulsatile shear stress in endothelial cells[J]. Proc Natl Acad Sci U S A, 2021, 118(21) : e2103982118.
[32]	Miao Z F, Adkins-Threats M, Burclaff J R, et al. A Metformin-Responsive Metabolic Pathway Controls Distinct Steps in Gastric Progenitor Fate Decisions and Maturation[J]. Cell Stem Cell, 2020, 26(6): 910-925.e6.
[33]	Liu L, Cheng Y, Wang J, et al. Simulated Microgravity Suppresses Osteogenic Differentiation of Mesenchymal Stem Cells by Inhibiting Oxidative Phosphorylation[J]. Int J Mol Sci, 2020, 21(24) : 9747.
[34]	Hao Y, Wu M, Wang J. Fibroblast growth factor-2 ameliorates tumor necrosis factor-alpha-induced osteogenic damage of human bone mesenchymal stem cells by improving oxidative phosphorylation[J]. Mol Cell Probes, 2020, 52: 101538.

Preliminary study on the mechanism of KLF4 regulation on energy metabolism in human periodontal ligament stem cells under high glucose inflammatory environment

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