HSP60 is involved in the neuroprotective effects of naloxone

Cheng,Wenjing; Li,Yunhong; Hou,Xiaolin; Zhang,Nan; Ma,Jiao; Ding,Feijia; Li,Fan; Miao,Zhenhua; Zhang,Yanli; Qi,Qi; Li,Guanghua; Shen,Ying; Liu,Juan; Huang,Weidong; Wang,Yin

doi:10.3892/mmr.2014.2411

HSP60 is involved in the neuroprotective effects of naloxone

Authors:
- Wenjing Cheng
- Yunhong Li
- Xiaolin Hou
- Nan Zhang
- Jiao Ma
- Feijia Ding
- Fan Li
- Zhenhua Miao
- Yanli Zhang
- Qi Qi
- Guanghua Li
- Ying Shen
- Juan Liu
- Weidong Huang
- Yin Wang
View Affiliations

Affiliations: Basic Medical College of Ningxia Medical University, Ningxia Key Laboratory of Cerebrocranial Diseases, Yinchuan, Ningxia 750004, P.R. China, The General Hospital of Ningxia Medical University, Yinchuan, Ningxia 750004, P.R. China, Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P.R. China
Published online on: July 22, 2014 https://doi.org/10.3892/mmr.2014.2411
Pages: 2172-2176

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Heat shock protein (HSP)60 is primarily a mitochondrial protein. Previous experiments have found that changes in the location of intracellular HSP60 have been associated with apoptosis. Extracellular HSP60 mediates apoptosis via its ligand, Toll‑like receptor (TLR)‑4. TLR‑4 is an important factor expressed on microglia, with a central role in generating neuroimmune responses in the pathogenesis of neurodegenerative disorders. Naloxone is a highly effective nonselective opioid receptor antagonist, and has been reported to be pharmacologically beneficial for the treatment of brain diseases through inhibiting microglia activation. However, the mechanisms underlying these beneficial effects of naloxone remain poorly understood. The present study aimed to investigate the role of HSP60 in the neuroprotective effects of naloxone on the production of proinflammatory mediators in lipopolysaccharide (LPS)‑stimulated BV2 murine microglial cells and the possible signaling pathways involved. The results demonstrated that naloxone significantly inhibited the expression and release of HSP60 in BV2 cells. The expression levels of heat shock factor (HSF)‑1 were upregulated in LPS‑activated BV2 cells, which indicated that the increased expression of HSP60 was driven by HSF‑1 activation. However, increased HSF‑1 levels may be downregulated by naloxone. The levels of TLR‑4 were elevated in activated BV2 cells, and then inhibited by naloxone. Activation of TLR‑4 is characterized by activation of nuclear factor‑κB (NF‑κB) followed by the production of various proinflammatory and neurotoxic factors. Data from the present study demonstrated that naloxone reduced the expression levels of NF‑κB and its upstream protein caspase‑3, and reduced the LPS‑induced production of nitric oxide, inducible nitric oxide synthase, tumor necrosis factor α, interleukin‑1β and interleukin‑6 in BV2 microglia. In light of this data, it was concluded that naloxone may exert its neuroprotective and anti‑inflammatory effects by inhibiting microglia activation through a HSP60‑TLR‑4‑NF‑κB signaling pathway.

View Figures

View References

1	Block ML, Zecca L and Hong JS: Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci. 8:57–69. 2007. View Article : Google Scholar : PubMed/NCBI
2	Hanisch UK and Kettenmann H: Microglia: active sensor and versatile effector cells in the normal and pathologic brain. Nat Neurosci. 10:1387–1394. 2007. View Article : Google Scholar : PubMed/NCBI
3	Gehrmann J, Matsumoto Y and Kreutzberg GW: Microglia: intrinsic immuneffector cell of the brain. Brain Res Rev. 20:269–287. 1995. View Article : Google Scholar : PubMed/NCBI
4	Innamorato NG, Lastres-Becker I and Cuadrado A: Role of microglial redox balance in modulation of neuroinflammation. Curr Opin Neurol. 22:308–314. 2009. View Article : Google Scholar : PubMed/NCBI
5	Lynch MA: The multifaceted profile of activated microglia. Mol Neurobiol. 40:139–156. 2009. View Article : Google Scholar : PubMed/NCBI
6	Li YH, Teng P, Wang Y, Zhang YM, Ma CJ, Pu J, et al: Expression and regulation of HSP60 in activated microglia cells. J Ningxia Med Coll. 8:712–714. 2011.
7	Zhang D, Sun L, Zhu H, Wang L, Wu W, Xie J and Gu J: Microglial LOX-1 reacts with extracellular HSP60 to bridge neuroinflammation and neurotoxicity. Neurochem Int. 61:1021–1035. 2012. View Article : Google Scholar : PubMed/NCBI
8	Lehnardt S, Schott E, Trimbuch T, Laubisch D, Krueger C, Wulczyn G, Nitsch R and Werber JR: 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. 2008. View Article : Google Scholar : PubMed/NCBI
9	Thanos S, Mey J and Wild M: Treatment of the adult retina with microglia-suppressing factors retards axotomy-induced neuronal degradation and enhances axonal regeneration in vivo and in vitro. J Neurosci. 13:455–466. 1993.
10	Thanos S: The Relationship of Microglial Cells to Dying Neurons During Natural Neuronal Cell Death and Axotomy-induced Degeneration of the Rat Retina. Eur J Neurosci. 3:1189–1207. 1991. View Article : Google Scholar
11	Teng P, Li YH, Cheng W, Zhou L, Shen Y and Wang Y: Neuroprotective effects of Lycium barbarum polysaccharides in lipopolysaccharide-induced BV2 microglial cells. Mol Med Rep. 7:1977–1981. 2013.PubMed/NCBI
12	Smith AP and Lee NM: Pharmacology of dynorphin. Annu Rev Pharmacol Toxicol. 28:123–140. 1988. View Article : Google Scholar
13	Faden AI and Salzman S: Pharmacological strategies in CNS trauma. Trends Pharmacol Sci. 13:29–35. 1992. View Article : Google Scholar : PubMed/NCBI
14	Fallis RJ, Fisher M and Lobo RA: A double blind trial of naloxone in the treatment of stroke. Stroke. 15:627–629. 1984. View Article : Google Scholar : PubMed/NCBI
15	Kan MN, Chen YT and Lee AY: Naloxone reversal of ischemic arrhythmia is stereospecific and suggests role of endogenous opioid peptides in ischemic heart disease. Proc Soc Exp Biol Med. 200:518–521. 1992. View Article : Google Scholar
16	Liu B, Du L and Hong JS: Naloxone protects rat dopaminergic neurons against inflammatory damage through inhibition of microglia activation and superoxide generation. J Pharmacol Exp Ther. 293:607–617. 2000.
17	Liu Y, Qin L, Wilson BC, An L, Hong JS and Liu B: Inhibition by naloxone stereoisomers of β-amyloid peptide (1–42)-induced superoxide production in microglia and degeneration of cortical and mesencephalic neurons. J Pharmacol Exp Ther. 302:1212–1219. 2002.
18	Kim SC, Stice JP, Chen L, Jung JS, Gupta S, Wang Y, Baumgarten G, Trial J and Knowlton AA: Extracellular heat shock protein 60, cardiac myocytes, and apoptosis. Circ Res. 105:1186–1195. 2009. View Article : Google Scholar : PubMed/NCBI
19	Lin L, Kim SC, Wang Y, Gupta S, et al: HSP60 in heart failure: abnormal distribution and role in cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol. 293:H2238–H2247. 2007. View Article : Google Scholar : PubMed/NCBI
20	Hansen JJ, Bross P, Westergaard M, Nielsen MN, Eiberg H, et al: Genomic structure of human mitochondrial chaperonin genes: HSP60 and HSP10 are localised head to head on chromosome 2 separated by a bidirectional promoter. Hum Genet. 112:71–77. 2003. View Article : Google Scholar
21	Aloisi F: Immune function of microglia. Glia. 36:165–179. 2001. View Article : Google Scholar : PubMed/NCBI
22	Kol A, Lichtman AH, Finberg RW, Libby P and Kurt-Jones EA: Cutting edge: heat shock protein (HSP) 60 activates the innate immune response: CD14 is an essential receptor for HSP60 activation of mononuclear cells. J Immunol. 164:13–17. 2000. View Article : Google Scholar : PubMed/NCBI
23	Wang Y, Chen L, Hagiwara N and Knowlton AA: Regulation of heat shock protein 60 and 72 expression in the failing heart. J Mol Cell Cardiol. 48:360–366. 2010. View Article : Google Scholar : PubMed/NCBI
24	Ohashi K, Burkart V, Flohé S and Kolb H: Cutting edge: heat shock protein 60 is a putative endogenous ligand of the toll-like receptor-4 complex. J Immunol. 164:558–561. 2000. View Article : Google Scholar : PubMed/NCBI
25	Chen J, Zhou Y, Mueller-Steiner S, Chen LF, Kwon H, Yi S, Mucke L and Gan L: SIRT1 protects against microglia-dependent amyloid-β toxicity through inhibiting NF-κB signaling. J Biol Chem. 280:40364–40374. 2005.PubMed/NCBI
26	Soria JA, Arroyo DS, Gaviglio EA, Rodriguez-Galan MC, Wang JM and Iribarren P: Interleukin 4 induces the apoptosis of mouse microglial cells by a caspase-dependent mechanism. Neurobiol Dis. 43:616–624. 2011. View Article : Google Scholar : PubMed/NCBI
27	Burguillos MA, Deierborg T, Kavanagh E, Persson A, Hajji N, Garcia-Quintanilla A, Cano J, Brundin P, Englund E, Venero JL and Joseph B: Caspase signalling controls microglia activation and neurotoxicity. Nature. 472:319–324. 2011. View Article : Google Scholar : PubMed/NCBI