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Hydroxysafflor Yellow A Attenuates Neuron Damage by Suppressing the Lipopolysaccharide-Induced TLR4 Pathway in Activated Microglial Cells

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Abstract

Microglia activation initiates a neurological deficit cascade that contributes to substantial neuronal damage and impairment following ischemia stroke. Toll-like receptor 4 (TLR4) has been demonstrated to play a critical role in this cascade. In the current study, we tested the hypothesis that hydroxysafflor yellow A (HSYA), an active ingredient extracted from Flos Carthami tinctorii, alleviated inflammatory damage, and mediated neurotrophic effects in neurons by inducing the TLR4 pathway in microglia. A non-contact Transwell co-culture system comprised microglia and neurons was treated with HSYA followed by a 1 mg/mL lipopolysaccharide (LPS) stimulation. The microglia were activated prior to neuronal apoptosis, which were induced by increasing TLR4 expression in the activated microglia. However, HSYA suppressed TLR4 expression in the activated microglia, resulting in less neuronal damage at the early stage of LPS stimulation. Western blot analysis and immunofluorescence indicated that dose-dependently HSYA down-regulated TLR4-induced downstream effectors myeloid differentiation factor 88 (MyD88), nuclear factor kappa b (NF-κB), and the mitogen-activated protein kinases (MAPK)-regulated proteins c-Jun NH2-terminal protein kinase (JNK), protein kinase (ERK) 1/2 (ERK1/2), p38 MAPK (p38), as well as the LPS-induced inflammatory cytokine release. However, HSYA up-regulated brain-derived neurotrophic factor (BDNF) expression. Our data suggest that HSYA could exert neurotrophic and anti-inflammatory functions in response to LPS stimulation by inhibiting TLR4 pathway-mediated signaling.

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References

  • Barbierato M, Facci L, Argentini C, Marinelli C, Skaper SD, Giusti P (2015) Astrocyte-microglia cooperation in the expression of a pro-inflammatory phenotype. CNS Neurol Disord 201312(5):608–618

    Google Scholar 

  • Barton GM, Medzhitov R (2003) Toll-like receptor signaling pathways. Science 300:1524–1525

    Article  CAS  PubMed  Google Scholar 

  • Block ML, Zecca L, Hong JS (2007) Microglia-mediated neuro-toxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 8:57–69

    Article  CAS  PubMed  Google Scholar 

  • Bozic I, Savic D, Laketa D, Bjelobaba I, Milenkovic I, Pekovic S, Nedeljkovic N, Lavrnja I (2015) Benfotiamine attenuates inflammatory response in LPS stimulated BV-2 microglia. PLoS One 10:e0118372

    Article  PubMed  PubMed Central  Google Scholar 

  • Buchanan MM, Hutchinson M, Watkins LR, Yin H (2010) Toll-like receptor 4 in CNS pathologies. J Neurochem 114:13–27

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chen L, Xiang Y, Kong L, Zhang X, Sun B, Wei X, Liu H (2013) Hydroxysafflor yellow A protects against cerebral ischemia-reperfusion injury by anti-apoptotic effect through PI3 K/Akt/GSK3β pathway in rat. Neurochem Res 38:2268–2275

    Article  PubMed  Google Scholar 

  • Chen L, Yang Y, Li CT, Zhang SR, Zheng W, Wei EQ, Zhang LH (2015) CysLT2 receptor mediates lipopolysaccharide-induced microglial inflammation and consequent neurotoxicity in vitro. Brain Res 1624:433–445

    Article  CAS  PubMed  Google Scholar 

  • de Bernardo S, Canals S, Casarejos MJ, Rodriguez-Martin E, Mena MA (2003) Glia-conditioned medium induces de novo synthesis of tyrosine hydroxylase and increases dopamine cell survival by differential signaling pathways. J Neurosci Res 73:818–830

    Article  PubMed  Google Scholar 

  • Doeppner TR, Kaltwasser B, Teli MK, Bretschneider E, Bahr M, Hermann DM (2014) Effects of acute versus post-acute systemic delivery of neural progenitor cells on neurological recovery and brain remodeling after focal cerebral ischemia in mice. Cell Death Dis 5:e1386

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • He Y, Zhou A, Jiang W (2013) Toll-like receptor 4-mediated signaling participates in apoptosis of hippocampal neurons. Neural Regen Res 8:2744–2753

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hines DJ, Choi HB, Hines RM, Phillips AG, MacVicar BA (2013) Prevention of LPS-induced microglia activation, cytokine production and sickness behavior with TLR4 receptor interfering peptides. PLoS One 8:e60388

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ji DB, Zhang LY, Li CL, Ye J, Zhu HB (2009) Effect of Hydroxysafflor yellow A on human umbilical vein endothelial cells under hypoxia. Vascul Pharmacol 50:137–145

    Article  CAS  PubMed  Google Scholar 

  • Jiang Z, Jiang JX, Zhang GX (2014) Macrophages: a double-edged sword in experimental autoimmune encephalomyelitis. Immunol Lett 160:17–22

    Article  CAS  PubMed  Google Scholar 

  • Kawamura K, Kawamura N, Kawagoe Y, Kumagai J, Fujimoto T, Terada Y (2012) Suppression of hydatidiform molar growth by inhibiting endogenous brain-derived neurotrophic factor/tyrosine kinase B signaling. Endocrinology 153:3972–3981

    Article  CAS  PubMed  Google Scholar 

  • Kettenmann H, Hanisch UK, Noda M, Verkhratsky A (2011) Physiology of microglia. Physiol Rev 91:461–553

    Article  CAS  PubMed  Google Scholar 

  • Klintworth H, Garden G, Xia Z (2009) Rotenone and paraquat do not directly activate microglia or induce inflammatory cytokine release. Neurosci Lett 62:1–5

    Article  Google Scholar 

  • Kraft AD, Harry GJ (2011) Features of microglia and neuroinflammation relevant to environmental exposure and neurotoxicity. Int J Environ Res Public Health 8:2980–3018

    Article  PubMed  PubMed Central  Google Scholar 

  • Kurpius D, Wilson N, Fuller L, Hoffman A, Dailey ME (2006) Early activation, motility, and homing of neonatal microglia to injured neurons does not require protein synthesis. Glia 54:58–70

    Article  PubMed  Google Scholar 

  • Laflamme N, Rivest S (2001) Toll-like receptor 4: the missing link of the cerebral innate immune response triggered by circulating gram-negative bacterial cell wall components. FASEB J 15:155–163

    Article  CAS  PubMed  Google Scholar 

  • Lee EJ, Ko HM, Jeong YH, Park EM, Kim HS (2015a) β-Lapachone suppresses neuroinflammation by modulating the expression of cytokines and matrix metalloproteinases in activated microglia. Mol Neurodegener 10:61

    Article  Google Scholar 

  • Lee KM, Bang J, Kim BY, Lee IS, Han JS, Hwang BY, Jeon WK (2015b) Fructus mume alleviates chronic cerebral hypoperfusion-induced white matter and hippocampal damage via inhibition of inflammation and downregulation of TLR4 and p38 MAPK signaling. BMC Complement Altern Med 15:125

    Article  PubMed  PubMed Central  Google Scholar 

  • Lehnardt S (2010) Innate immunity and neuroinflammation in the CNS: the role of microglia in Toll-like receptor-mediated neuronal injury. Glia 58:253–263

    PubMed  Google Scholar 

  • Lehnardt S, Massillon L, Follett P, Jensen FE, Ratan R, Rosenberg PA, Volpe JJ, Vartanian T (2003) Activation of innate immunity in the CNS triggers neurodegeneration through a Toll-like receptor 4-dependent pathway. Proc Natl Acad Sci USA 100:8514–8519

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lehnardt S, Schott E, Trimbuch T, Laubisch D, Krueger C, Wulczyn G, Nitsch R, Weber JR (2008) A vicious cycle involving release of heat shock protein 60 from injured cells and activation of toll-like receptor 4 mediates neurodegeneration in the CNS. J Neurosci 28:2320–2331

    Article  CAS  PubMed  Google Scholar 

  • Li GZ, Zhang Y, Zhao JB, Wu GJ, Su XF, Hang CH (2011) Expression of myeloid differentiation primary response protein 88 (Myd88) in the cerebral cortex after experimental traumatic brain injury in rats. Brain Res 1396:96–104

    Article  CAS  PubMed  Google Scholar 

  • Li J, Zhang S, Lu M, Chen Z, Chen C, Han L, Zhang M, Xu Y (2013) Hydroxysafflor yellow A suppresses inflammatory responses of BV2 microglia after oxygen-glucose deprivation. Neurosci Lett 535:51–56

    Article  CAS  PubMed  Google Scholar 

  • Lindvall O, Ernfors P, Bengzon J, Kokaia Z, Smith ML, Siesjö BK, Persson H (1992) Differential regulation of mRNAs for nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3 in the adult rat brain following cerebral ischemia and hypoglycemic coma. Proc Natl Acad Sci USA 89:648–652

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lv Y, Qian Y, Fu L, Chen X, Zhong H, Wei X (2015) Hydroxysafflor yellow A exerts neuroprotective effects in cerebral ischemia reperfusion-injured mice by suppressing the innate immune TLR4-inducing pathway. Eur J Pharmacol 769:324–332

    Article  CAS  PubMed  Google Scholar 

  • Madeddu F, Naska S, Bozzi Y (2004) BDNF down-regulates the caspase 3 pathway in injured geniculo-cortical neurones. NeuroReport 15:2045–2049

    Article  CAS  PubMed  Google Scholar 

  • Martorana F, Guidotti G, Brambilla L, Rossi D, Withaferin A (2015) Inhibits nuclear factor-κB-dependent pro-inflammatory and stress response pathways in the astrocytes. Neural Plast 2015:381964

    PubMed  PubMed Central  Google Scholar 

  • Morrison HW, Filosa JA (2013) A quantitative spatiotemporal analysis of microglia morphology during ischemic stroke and reperfusion. J Neuroinflammation 10:4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nakajima K, Kohsaka S (2004) Microglia: neuroprotective and neurotrophic cells in the central nervous system. Curr Drug Targets Cardiovasc Haematol Disord 4:65–84

    Article  CAS  PubMed  Google Scholar 

  • Pardo B, Paino CL, Casarejos MJ, Mena MA (1997) Neuronal-enriched cultures from embryonic rat ventral mesencephalon for pharmacological studies of dopamine neurons. Brain Res Brain Res Protocol 1:127–132

    Article  CAS  Google Scholar 

  • Park J, Min JS, Kim B, Chae UB, Yun JW, Choi MS, Kong IK, Chang KT, Lee DS (2015) Mitochondrial ROS govern the LPS-induced pro-inflammatory response in microglia cells by regulating MAPK and NF-κB pathways. Neurosci Lett 584:191–196

    Article  CAS  PubMed  Google Scholar 

  • Prathab Balaji S, Vijay Chand C, Justin A, Ramanathan M (2015) Telmisartan mediates anti-inflammatory and not cognitive function through PPAR-γ agonism via SARM and MyD88 signaling. Pharmacol Biochem Behav 137:60–68

    Article  CAS  PubMed  Google Scholar 

  • Shulga A, Blaesse A, Kysenius K, Huttunen HJ, Tanhuanpää K, Saarma M, Rivera C (2009) Thyroxin regulates BDNF expression to promote survival of injured neurons. Mol Cell Neurosci 42:408–418

    Article  CAS  PubMed  Google Scholar 

  • Song M, Jin J, Lim JE, Kou J, Pattanayak A, Rehman JA, Kim HD, Tahara K, Lalonde R, Fukuchi K (2011) TLR4 mutation reduces microglial activation, increases Aβ deposits and exacerbates cognitive deficits in a mouse model of Alzheimer’s disease. J Neuroinflammation 8:92

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song J, Cheon SY, Jung W, Lee WT, Lee JE (2014) Resveratrol induces the expression of interleukin-10 and brain-derived neurotrophic factor in BV2 microglia under hypoxia. Int J Mol Sci 15:15512–15529

    Article  PubMed  PubMed Central  Google Scholar 

  • Sun CY, Pei CQ, Zang BX, Wang L, Jin M (2010) The ability of hydroxysafflor yellow a to attenuate lipopolysaccharide-induced pulmonary inflammatory injury in mice. Phytother Res 24:1788–1795

    Article  CAS  PubMed  Google Scholar 

  • Tanaka S, Ide M, Shibutani T, Ohtaki H, Numazawa S, Shioda S, Yoshida T (2006) Lipopolysaccharide-induced microglial activation induces learning and memory deficits without neuronal celldeath in rats. J Neurosci Res 83:557–566

    Article  CAS  PubMed  Google Scholar 

  • Tao X, Sun X, Yin L, Han X, Xu L, Qi Y, Xu Y, Li H, Lin Y, Liu K, Peng J (2015) Dioscin ameliorates cerebral ischemia/reperfusion injury through the downregulation of TLR4signaling via HMGB-1 inhibition. Free Radic Biol Med 84:103–115

    Article  CAS  PubMed  Google Scholar 

  • Tian J, Li G, Liu Z, Fu F (2008) Hydroxysafflor yellow A inhibits rat brain mitochondrial permeability transition pores by a free radical scavenging action. Pharmacology 82:121–126

    Article  CAS  PubMed  Google Scholar 

  • Umschweif G, Shabashov D, Alexandrovich AG, Trembovler V, Horowitz M, Shohami E (2014) Neuroprotection after traumatic brain injury in heat-acclimated mice involves induced neurogenesis and activation of angiotensin receptor type 2 signaling. J Cereb Blood Flow Metab 34:1381–1390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Walker PA, Shah SK, Jimenez F, Aroom KR, Harting MT, Cox CS Jr (2012) Bone marrow-derived stromal cell therapy for traumatic brain injury is neuroprotective via stimulation of non-neurologic organ systems. Surgery 152:790–793

    Article  PubMed  Google Scholar 

  • Wang X, Stridh L, Li W, Dean J, Elmgren A, Gan L, Eriksson K, Hagberg H, Mallard C (2009) Lipopolysaccharide sensitizes neonatal hypoxic-ischemic brain injury in a MyD88-dependent manner. J Immunol 183:7471–7477

    Article  CAS  PubMed  Google Scholar 

  • Wang W, Wang T, Feng WY, Wang ZY, Cheng MS, Wang YJ (2014) Ecdysterone protects gerbil brain from temporal global cerebral ischemia/reperfusion injury via preventing neuron apoptosis and deactivating astrocytes and microglia cells. Neurosci Res 81–82:21–29

    Article  PubMed  Google Scholar 

  • Wu Y, Wang L, Jin M, Zang BX (2012) Hydroxysafflor yellow A alleviates early inflammatory response of bleomycin-induced mice lung injury. Biol Pharm Bull 35:515–522

    Article  PubMed  Google Scholar 

  • Wu B, Huang Y, Braun AL, Tong Z, Zhao R, Li Y, Liu F, Zheng JC (2015) Glutaminase-containing microvesicles from HIV-1-infected macrophages and immune-activated microglia induce neurotoxicity. J Neuroinflammation 12:133

    Article  Google Scholar 

  • Yao L, Kan EM, Lu J, Hao A, Dheen ST, Kaur C, Ling EA (2013) Toll-like receptor 4 mediates microglial activation and production of inflammatory mediators in neonatal rat brain following hypoxia: role of TLR4 in hypoxic microglia. J Neuroinflammation 10:23

    Article  PubMed  PubMed Central  Google Scholar 

  • Yoo KY, Yoo DY, Hwang IK, Park JH, Lee CH, Choi JH, Kwon SH, Her S, Lee YL, Won MH (2011) Time-course alterations of Toll-like receptor 4 and NF-κB p65, and their co-expression in the gerbil hippocampal CA1 region after transient cerebral ischemia. Neurochem Res 36:2417–2426

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Zeng L, Yu T, Xu Y, Pu S, Du D, Jiang W (2014a) Positive feedback loop of autocrine BDNF from microglia causes prolonged microglia activation. Cell Physiol Biochem 34:715–723

    Article  PubMed  Google Scholar 

  • Zhang Z, Wu Z, Zhu X, Hui X, Pan J, Xu Y (2014b) Hydroxy-safflor yellow A inhibits neuroinflammation mediated by Aβ1–42 in BV-2 cells. Neurosci Lett 562:39–44

    Article  CAS  PubMed  Google Scholar 

  • Zhang ZH, Yu LJ, Hui XC, Wu ZZ, Yin KL, Yang H, Xu Y (2014c) Hydroxy-safflor yellow A attenuates Aβ1–42-induced inflammation by modulating the JAK2/STAT3/NF-κB pathway. Brain Res 1563:72–80

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Li YJ, Wu XY, Hong Z, Wei WS (2015) MicroRNA-181c negatively regulates the inflammatory response in oxygen-glucose-deprived microglia by targeting Toll-like receptor 4. J Neurochem 132:713–723

    Article  CAS  PubMed  Google Scholar 

  • Zhu HT, Bian C, Yuan JC, Chu WH, Xiang X, Chen F, Wang CS, Feng H, Lin JK (2014) Curcumin attenuates acute inflammatory injury by inhibiting the TLR4/MyD88/NF-κB signaling pathway in experimental traumatic brain injury. J Neuroinflammation 11:59

    Article  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Pharmacy Department, The First Affiliated Hospital of Nanchang University, Yongwai Street 17, Nanchang, 330006, China

    Yanni Lv, Aijun Ou-yang & Longsheng Fu

  2. Institute of Translational Medicine, Nanchang University, Nanchang, 330001, China

    Yisong Qian

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  1. Yanni Lv
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  2. Yisong Qian
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  3. Aijun Ou-yang
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  4. Longsheng Fu
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Correspondence to Yanni Lv.

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Lv, Y., Qian, Y., Ou-yang, A. et al. Hydroxysafflor Yellow A Attenuates Neuron Damage by Suppressing the Lipopolysaccharide-Induced TLR4 Pathway in Activated Microglial Cells. Cell Mol Neurobiol 36, 1241–1256 (2016). https://doi.org/10.1007/s10571-015-0322-3

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