棕榈酸促进脂多糖进入小胶质细胞并引发凋亡和焦亡

冯玉虎  杨衍卓  吕海燕  郁清婷  杨最素  袁法磊

doi:10.16098/j.issn.0529-1356.2025.04.004

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解剖学报 ›› 2025, Vol. 56 ›› Issue (4) : 404-412. DOI: 10.16098/j.issn.0529-1356.2025.04.004

神经生物学

棕榈酸促进脂多糖进入小胶质细胞并引发凋亡和焦亡

冯玉虎¹ 杨衍卓¹ 吕海燕² 郁清婷¹ 杨最素¹ 袁法磊^1*

作者信息 +

Palmitic acid increasing the entry of lipopolysaccharide into microglial cytosol and eliciting pyroptosis and apoptosis

FENG Yu-hu1 YANG Yan-zhuo¹ Lü Hai-yan² YU Qing-ting¹ YANG Zui-su¹ YUAN Fa-lei^1*

Author information +

文章历史 +

摘要

目的探究棕榈酸（PA）与脂多糖（LPS）相互作用后引发的小胶质细胞死亡的类型和机制。方法将BV-2小胶质细胞分为3组研究细胞凋亡：牛血清清蛋白 (BSA) 组、PA处理组、星形孢菌素（STA）组；另外分为4组研究细胞坏死：BSA组、BSA+抑制剂组、PA组、PA+抑制剂组。Western blotting分析凋亡和坏死通路关键蛋白表达水平。通过饲喂美国癌症研究所（ICR）小鼠高脂饮食验证PA对小胶质细胞的作用。结果在BSA组和PA组中的小胶质细胞均发生了凋亡，PA激活了Caspase-3、Caspase-7以及多聚ADP-核糖聚合酶（PARP），然而STA组与BSA组相比，活化的Caspase-7水平变化不显著。抑制铁死亡、坏死性凋亡或自噬都不能保护PA引起的细胞损伤，而Caspase-11的抑制剂蟛蜞菊内酯（WE）可以显著改善PA所引起的细胞损伤。本研究还发现，PA可以促进LPS进入小胶质细胞并引发焦亡。此现象以及WE的保护作用进一步在高脂饮食小鼠模型中通过免疫荧光染色和Western blotting得到证实。结论 PA诱导小胶质细胞发生凋亡和焦亡，而PA同时可以促进LPS进入小胶质细胞并引发焦亡。

Abstract

Objective To investigate the types and mechanisms of microglial cell death induced by interaction between palmitic acid (PA) and lipopolysaccharide (LPS). Methods BV-2 microglial cells were divided into three groups for apoptosis research, BSA group, PA treatment group, and staurosporine (STA) group. They were further divided into four groups for necrosis research, BSA group, BSA + inhibitor group, PA group, and PA + inhibitor group. Western blotting was conducted to assess the expression levels of key proteins involved in apoptosis and necrosis pathways. The effect of PA on microglial cells was validated through feeding a high-fat diet to Institute of Cancer Research(ICR) mice. Results Apoptotic microglia were observed in both BSA group and PA group, PA significantly induced the activation of caspase-3, caspase-7, and poly ADP-ribose polymerase(PARP). However, compared to the BSA group, the level of activated Caspase-7 in the STA group did not change significantly. Inhibition of ferroptosis, necroptosis, or autophagy did not protect against PA-induced cell damage, while the Caspase-11 inhibitor, wedelolactone (WE), significantly improved PA induced cell damage. This study also found that PA could promote LPS entry into microglial cells and induce pyroptosis. This phenomenon and the protective effect of WE were further confirmed in a high-fat diet mouse model through immunofluorescent staining and Western blotting. Conclusion PA induces apoptosis and pyroptosis in microglial cells, while simultaneously promoting LPS entry into microglial cells and inducing pyroptosis.

导出引用

冯玉虎杨衍卓吕海燕郁清婷杨最素袁法磊. 棕榈酸促进脂多糖进入小胶质细胞并引发凋亡和焦亡[J]. 解剖学报. 2025, 56(4): 404-412 https://doi.org/10.16098/j.issn.0529-1356.2025.04.004

FENG Yu-hu YANG Yan-zhuo Lü Hai-yan YU Qing-ting YANG Zui-su YUAN Fa-lei. Palmitic acid increasing the entry of lipopolysaccharide into microglial cytosol and eliciting pyroptosis and apoptosis[J]. Acta Anatomica Sinica. 2025, 56(4): 404-412 https://doi.org/10.16098/j.issn.0529-1356.2025.04.004

中图分类号： R361+.2

参考文献

［1］Stumvoll M, Nurjhan N, Perriello G, et al. Metabolic effects of metformin in non-insulin-dependent diabetes mellitus［J］. N Engl J Med, 1995, 333(9):550-554.

［2］Kang R, Zeng L, Zhu S, et al. Lipid peroxidation drives gasdermin D-mediated pyroptosis in lethal polymicrobial sepsis［J］. Cell Host Microbe, 2018, 24(1):97-108.e104.

［3］Waise TMZ, Toshinai K, Naznin F, et al. One-day high-fat diet induces inflammation in the nodose ganglion and hypothalamus of mice［J］. Biochem Biophys Res Commun, 2015, 464(4):1157-1162.

［4］Alsabeeh N, Chausse B, Kakimoto PA, et al. Cell culture models of fatty acid overload: problems and solutions［J］. Biochim Biophys Acta Mol Cell Biol Lipids, 2018, 1863(2):143-151.

［5］Fricker M, Tolkovsky AM, Borutaite V, et al. Neuronal cell death［J］. Physiol Rev, 2018, 98(2):813-880.

［6］Wang XZh, Yuan JY, Zhang J, et al. Role of inflammation between pyroptosis and atherosclerosis［J］. Acta Anatomica Sinica, 2019, 50(4):543-548.(in Chinese)

王熹芝,袁佳莹,张剑,等. 炎症反应在细胞焦亡和动脉粥样硬化之间的作用［J］.解剖学报,2019,50(4):543-548.

［7］Kapralov AA, Yang Q, Dar HH, et al. Redox lipid reprogramming commands susceptibility of macrophages and microglia to ferroptotic death［J］. Nat Chem Biol, 2020, 16(3):278-290.

［8］Qin S, Yang C, Huang W, et al. Sulforaphane attenuates microglia-mediated neuronal necroptosis through down-regulation of MAPK/NF-κB signaling pathways in LPS-activated BV-2 microglia［J］. Pharmacol Res, 2018, 133(8):218-235.

［9］Wang Z, Wang G, Wang Y, et al. Omp31 of brucella inhibits NF-κB p65 signaling pathway by inducing autophagy in BV-2 microglia［J］. Neurochem Res, 2021, 46(12):3264-3272.

［10］Herr DR, Yam TYA, Tan WSD, et al. Ultrastructural characteristics of DHA-induced pyroptosis［J］. Neuromolecular Med, 2020, 22(2):293-303.

［11］Nakandakari SCBR, Mu?oz VR, Kuga GK, et al. Short-term high-fat diet modulates several inflammatory, ER stress, and apoptosis markers in the hippocampus of young mice［J］. Brain Behav Immun, 2019, 79(9):284-293.

［12］Wang N, Ma H, Li J, et al. HSF1 functions as a key defender against palmitic acid-induced ferroptosis in cardiomyocytes［J］. J Mol Cell Cardiol, 2021, 150(6):65-76.

［13］Tao L, Yi Y, Chen Y, et al. RIP1 kinase activity promotes steatohepatitis through mediating cell death and inflammation in macrophages［J］. Cell Death Differ, 2021, 28(4):1418-1433.

［14］Park S, Oh TS, Kim S, et al. Palmitate-induced autophagy liberates monounsaturated fatty acids and increases AGRP expression in hypothalamic cells［J］. Anim Cells Syst (Seoul), 2019, 23(6):384-391.

［15］Zeng X, Zhu M, Liu X, et al. Oleic acid ameliorates palmitic acid induced hepatocellular lipotoxicity by inhibition of ER stress and pyroptosis［J］. Nutr Metab (Lond), 2020, 17(7):11.

［16］Srikanth M, Chandrasaharan K, Zhao X, et al. Metabolism of docosahexaenoic acid (DHA) induces pyroptosis in BV-2 microglial cells［J］. Neuromolecular Med, 2018, 20(4):504-514.

［17］Hu Y, Wang B, Li S, et al. Pyroptosis, and its role in central nervous system disease［J］. J Mol Biol, 2022, 434(4):167379.

［18］Kobori M, Yang Z, Gong D, et al. Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex［J］. Cell Death Differ, 2004, 11(1):123-130.

［19］Kayagaki N, Warming S, Lamkanfi M, et al. Non-canonical inflammasome activation targets caspase-11［J］. Nature, 2011, 479(7371):117-121.

［20］Lee J, Hur J, Lee P, et al. Dual role of inflammatory stimuli in activation-induced cell death of mouse microglial cells. Initiation of two separate apoptotic pathways via induction of interferon regulatory factor-1 and caspase-11［J］. J Biol Chem, 2001, 276(35):32956-32965.

［21］Nozaki K, Maltez VI, Rayamajhi M, et al. Caspase-7 activates ASM to repair gasdermin and perforin pores［J］. Nature, 2022, 606(7916):960-967.

［22］LI HM, Peng M, Yang QQ, et al. Pro-apoptosis effects of docosahexaenoic acid on rat C6 glioma cells in vitro［J］. Acta Anatomica Sinica, 2018, 49(4): 419-423. (in Chinese)

李浩明,彭敏,杨清清,等.二十二碳六烯酸对体外大鼠C6胶质瘤细胞的促凋亡作用［J］.解剖学报,2018,49(4):419-423.

［23］Tewari M, Quan LT, O’Rourke K, et al. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase［J］. Cell, 1995, 81(5):801-809.

［24］Gobeil S, Boucher CC, Nadeau D, et al. Characterization of the necrotic cleavage of poly(ADP-ribose) polymerase (PARP-1): implication of lysosomal proteases［J］. Cell Death Differ, 2001, 8(6):588-594.

［25］Degterev A, Huang Z, Boyce M, et al. Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury［J］. Nat Chem Biol, 2005, 1(2):112-119.

［26］Li H, Xiao Y, Tang L, et al. Adipocyte fatty acid-binding protein promotes palmitate-induced mitochondrial dysfunction and apoptosis in macrophages［J］. Front Immunol, 2018, 9(5):81.

［27］Kayagaki N, Kornfeld OS, Lee BL, et al. NINJ1 mediates plasma membrane rupture during lytic cell death［J］. Nature, 2021, 591(7848):131-136.

［28］Thaler JP, Yi CX, Schur EA, et al. Obesity is associated with hypothalamic injury in rodents and humans［J］. J Clin Invest, 2012, 122(1):153-162.

［29］Graeber MB, Li W, and Rodriguez ML. Role of microglia in CNS inflammation［J］. FEBS Lett, 2011, 585(23):3798-3805.

［30］Jia J, Claude-Taupin A, Gu Y, et al. Galectin-3 coordinates a cellular system for lysosomal repair and removal［J］. Dev Cell, 2020, 52(1):69-87.e68.

［31］Lo TH, Chen HL, Yao CI, et al. Galectin-3 promotes noncanonical inflammasome activation through intracellular binding to lipopolysaccharide glycans［J］. Proc Natl Acad Sci USA, 2021, 118(30):116-123.

［32］Zusso M, Lunardi V, Franceschini D, et al. Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway［J］. J Neuroinflammation, 2019, 16(1):148.

［33］Deng M, Tang Y, Li W, et al. The endotoxin delivery protein hmgb1 mediates caspase-11-dependent lethality in sepsis［J］. Immunity, 2018, 49(4):740-753.e747.

［34］Golec M. Cathelicidin LL-37: LPS-neutralizing, pleiotropic peptide［J］. Ann Agric Environ Med, 2007, 14(1):1-4.

［35］Kang SJ, Wang S, Hara H, et al. Dual role of caspase-11 in mediating activation of caspase-1 and caspase-3 under pathological conditions［J］. J Cell Biol, 2000, 149(3):613-622.

［36］Rogers C, Fernandes-Alnemri T, Mayes L, et al. Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death［J］. Nat Commun, 2017, 8(8):14128.