特女贞苷对脂多糖诱导的巨噬细胞促炎反应的抑制作用及机制

doi:10.3969/j.issn.1004-616x.2023.04.001

Abstract

Abstract: OBJECTIVE： This study aimed to investigate effects of nuezhenide (NED) on LPS-induced proinflammatory response in macrophages. METHODS： RAW264.7 cells were treated with 50 μmol/L NED for 12 hours. Differential gene expressions were detected via transcriptome，cell viability using the CCK8 kit，and apoptosis rates measured using Annexin V-FITC/PI staining，and expression and localization of Nrf2 protein were detected by immunofluorescence. The cells were treated with LPS (100 ng/mL) and NED (12.5，25 and 50 μmol/L) for 12 hours. Then，TNF-α and IL-6 contents in the supernatant of cultured cells were determined by ELISA，and expressions of TNF-α and IL-6 by RT-PCR. Moreover，ROS，mitochondrial mass，and membrane potential were measured by flow cytometry after staining with DCFH-DA，MitoTracker-Green and JC-1，respectively. Enzyme activities of SOD and CAT，activities of mitochondrial complexes I and Ⅲ，as well as ATP contents were determined using commercial kits. RESULTS： Our results show that 50 μmol/L NED had no significant effects on survival and apoptosis of the RAW264.7 macrophages (P>0.05). However，NED treatments blunted LPS-induced expression of TNF-α and IL-6 (P<0.05)，and enhanced expression of anti-inflammatory cytokines (P<0.05). Meanwhile，NED significantly promoteed Nrf2 nuclear translocation and downstream antioxidant transcription (P<0.05)，which contributed to inhibition of LPS-induced accumulation of ROS and GSSG (P<0.05). In addition，NED reversed the LPS-induced decreased of mitochondrial membrane potential，mitochondrial complexes I and III activities，as well as ATP contents through promoting mitochondrial biogenesis (P<0.05). CONCLUSION： NED inhibited LPS-induced macrophages pro-inflammatory differentiation through enhancing Nrf2 antioxidant system and mitochondrial biogenesis. Application of NED may be a potential strategy for treatment of inflammatory diseases.

Key words: nuezhenide, macrophages, inflammatory response, antioxidant, mitochondrial biogenesis

CLC Number:

R392.11

JIANG Xiaoxu, SUN Muhan, ZHANG Boxu, WANG Yuyue, MA Jiajun, SHI Minjie, YU Weihua, GUO Xian. Protection against LPS-induced pro-inflammatory response by nuezhenide in macrophages[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(4): 245-252.

References

[1] MCINNES I B, GRAVALLESE E M. Immune-mediated inflammatory disease therapeutics: past, present and future[J]. Nat Rev Immunol, 2021, 21(10): 680-686.
[2] EIJGELAAR W J, HORREVOETS A J, BIJNENS A P, et al. Equivalence testing in microarray analysis: similarities in the transcriptome of human atherosclerotic and nonatherosclerotic macrophages[J]. Physiol Genomics, 2010, 41(3): 212-223.
[3] KIM A J, XU N, YUTZEY K E. Macrophage lineages in heart valve development and disease[J]. Cardiovasc Res, 2021, 117(3): 663-673.
[4] FACCHIN B M, DOS REIS G O, VIEIRA G N, et al. Inflammatory biomarkers on an LPS-induced RAW 264.7 cell model: a systematic review and meta-analysis[J]. Inflamm Res, 2022, 71(7/8): 741-758.
[5] YU W H, WANG X, ZHAO J Z, et al. Stat2-Drp1 mediated mitochondrial mass increase is necessary for pro-inflammatory differentiation of macrophages[J]. Redox Biol, 2020, 37: 101761.
[6] SCHJERNING A M, MCGETTIGAN P, GISLASON G. Cardiovascular effects and safety of (non-aspirin) NSAIDs[J]. Nat Rev Cardiol, 2020, 17(9): 574-584.
[7] RIBEIRO H, RODRIGUES I, NAPOLE-O L, et al. Non-steroidal anti-inflammatory drugs (NSAIDs), pain and aging: adjusting prescription to patient features[J]. Biomedecine Pharmacother, 2022, 150: 112958.
[8] 刘美红, 邹峥嵘. 女贞子化学成分、药理作用及药动学研究进展[J]. 热带亚热带植物学报, 2022, 30(3): 446-460.
[9] 龙玉婷, 马紫童, 刘仁慧. 女贞子延缓衰老作用机制研究现状[J]. 中国中医药信息杂志, 2022, 29(6): 139-142.
[10] 张明发, 沈雅琴. 女贞子抗炎、抗肿瘤和免疫调节作用的研究进展[J]. 现代药物与临床, 2012, 27(5): 536-542.
[11] 胡冬梅, 陆杨, 房敏峰, 等. 特女贞苷对四氯化碳致小鼠急性肝损伤的保护作用[J]. 中国药理学通报, 2016, 32(9): 1260-1263.
[12] 顾闻, 刘特, 陈久林, 等. 特女贞苷对血管内皮细胞氧化损伤的作用研究[J]. 中国中西医结合杂志, 2018, 38(9): 1093-1098.
[13] 赵永见, 张伟强, 杨俊杰, 等. 早期应用不同剂量特女贞苷治疗肾性骨病小鼠骨质疏松的疗效研究[C]//第十四届中国实验动物科学年会论文集. 青岛, 2018: 577.
[14] 刘艳秋, 吴琪聪, 周家杰, 等. 女贞子成分对BMSC成骨分化及RAW264.7细胞破骨分化的影响研究[C]//世界中医药学会联合会中药新药创制专业委员会成立大会暨第一届学术年会论文集. 成都, 2016: 291-295.
[15] 战美, 邓雪, 朱迪, 等. 女贞子提取物及其成分促骨髓间充质干细胞成骨分化的作用[J]. 中华中医药杂志, 2018, 33(7): 2803-2806.
[16] GOODMAN S B, MARUYAMA M. Inflammation, bone healing and osteonecrosis: from bedside to bench[J]. J Inflamm Res, 2020, 13: 913-923.
[17] WANG Z C, ZHAO W Y, CAO Y Y, et al. The roles of inflammation in keloid and hypertrophic scars[J]. Front Immunol, 2020, 11: 603187.
[18] FEEHAN K T, GILROY D W. Is resolution the end of inflammation-[J]. Trends Mol Med, 2019, 25(3): 198-214.
[19] RATHINAM V A K, ZHAO Y, SHAO F. Innate immunity to intracellular LPS[J]. Nat Immunol, 2019, 20(5): 527-533.
[20] 姜晓旭, 郑义鹏, 赵九洲, 等. M1型巨噬细胞糖代谢重编程机制及其在炎症启动中的关键作用[J]. 癌变·畸变·突变, 2019, 31(1): 79-81, 85.
[21] JOSEFS T, BARRETT T J, BROWN E J, et al. Neutrophil extracellular traps promote macrophage inflammation and impair atherosclerosis resolution in diabetic mice[J]. JCI Insight, 2020, 5(7): e134796.
[22] 王笑妍, 李玫, 沈志纲, 等. 红景天苷药理作用研究进展[J]. 中成药, 2022, 44(12): 3932-3935.
[23] YU W H, ZHANG X D, WU H, et al. HO-1 is essential for tetrahydroxystilbene glucoside mediated mitochondrial biogenesis and anti-inflammation process in LPS-treated RAW264.7 macrophages[J]. Oxid Med Cell Longev, 2017, 2017: 1818575.
[24] LU S H, ZUO H J, HUANG J, et al. Chemical constituents from the leaves of Ligustrum robustum and their bioactivities[J]. Molecules, 2023, 28(1): 362.
[25] QIU Z C, ZHAO X X, WU Q C, et al. New secoiridoids from the fruits of Ligustrum lucidum[J]. J Asian Nat Prod Res, 2018, 20(5): 431-438.
[26] KARANIKAS E, VLASIDOU D I. Psychologically traumatic stress inducing redox - inflammation interplay；which comes first-[J]. Pharmacol Biochem Behav, 2021, 209: 173254.
[27] PETERS A, NAWROT T S, BACCARELLI A A. Hallmarks of environmental insults[J]. Cell, 2021, 184(6): 1455-1468.
[28] WEIGERT A, VON KNETHEN A, FUHRMANN D, et al. Redox-signals and macrophage biology[J]. Mol Aspects Med, 2018, 63: 70-87.
[29] PéREZ S, RIUS-PéREZ S. Macrophage polarization and reprogramming in acute inflammation: a redox perspective[J]. Antioxidants, 2022, 11(7): 1394.
[30] ROJO DE LA VEGA M, CHAPMAN E, ZHANG D D. NRF2 and the hallmarks of cancer[J]. Cancer Cell, 2018, 34(1): 21-43.
[31] TONELLI C, CHIO I I C, TUVESON D A. Transcriptional regulation by Nrf2[J]. Antioxid Redox Signal, 2018, 29(17): 1727-1745.
[32] ULASOV A V, ROSENKRANZ A A, GEORGIEV G P, et al. Nrf2/Keap1/ARE signaling: towards specific regulation[J]. Life Sci, 2022, 291: 120111.
[33] V GANESH G, GANESAN K, XU B J, et al. Nrf2 driven macrophage responses in diverse pathophysiological contexts: disparate pieces from a shared molecular puzzle[J]. Biofactors, 2022, 48(4): 795-812.
[34] TU Y M, LIU J Z, KONG D Q, et al. Irisin drives macrophage anti-inflammatory differentiation via JAK2-STAT6-dependent activation of PPARγ and Nrf2 signaling[J]. Free Radic Biol Med, 2023, 201: 98-110.
[35] FORBES J M, THORBURN D R. Mitochondrial dysfunction in diabetic kidney disease[J]. Nat Rev Nephrol, 2018, 14(5): 291-312.
[36] CHAN D C. Mitochondrial dynamics and its involvement in disease[J]. Annu Rev Pathol, 2020, 15: 235-259.
[37] MARTíNEZ-REYES I, CHANDEL N S. Mitochondrial TCA cycle metabolites control physiology and disease[J]. Nat Commun, 2020, 11(1): 102.
[38] 康朦梦, 曾其毅. 脓毒症与线粒体生物发生的研究进展[J]. 广州医科大学学报, 2014, 42(5): 116-120.
[39] ZHANG M M, LIN J, WANG S J, et al. Melatonin protects against diabetic cardiomyopathy through Mst1/Sirt3 signaling[J]. J Pineal Res, 2017, 63(2). doi: 10.1111/jpi.12418.
[40] KOO S J, GARG N J. Metabolic programming of macrophage functions and pathogens control[J]. Redox Biol, 2019, 24: 101198.
[41] LANGSTON P K, SHIBATA M, HORNG T. Metabolism supports macrophage activation[J]. Front Immunol, 2017, 8: 61.

Protection against LPS-induced pro-inflammatory response by nuezhenide in macrophages

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