Repeated intra-nigrostriatal injection of phorbol myristate acetate induces microglial senescence in adult rats

Liu,Lin; Luo,Xiao‑Guang; Yu,Hong‑Mei; Feng,Yu; Ren,Yan; Yin,Ya‑Fu; Shang,Hong; He,Zhi‑Yi

doi:10.3892/mmr.2015.4412

Repeated intra-nigrostriatal injection of phorbol myristate acetate induces microglial senescence in adult rats

Authors:
- Lin Liu
- Xiao‑Guang Luo
- Hong‑Mei Yu
- Yu Feng
- Yan Ren
- Ya‑Fu Yin
- Hong Shang
- Zhi‑Yi He
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Affiliations: Department of Neurology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Nuclear Medicine, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Laboratory Medicine, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: October 1, 2015 https://doi.org/10.3892/mmr.2015.4412
Pages: 7271-7278
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Phorbol myristate acetate (PMA), as a potent tumor promoter, may induce microglial senescence. The present study investigated the effect of PMA infection on microglial senescence. From 58 male Sprague‑Dawley rats, 10 were randomly selected and divided into a PMA injection group, containing five rats (0.5 µg/µl PMA) and a control group, containing five rats (commensurable 0.9% saline). Immunofluorescent staining of Iba‑1 and enzyme‑linked immunosorbent assay analyses of the expression levels of tumor necrosis factor (TNF)‑α and interleukin (IL)‑1 β were performed in these two groups. The remaining 48 rats were randomly divided into the following three groups, each containing 16 rats: Repeated injection control group (commensurable normal saline, once a week for 4 weeks), single PMA injection group (0.5 µg/µl PMA, once in the first week) and repeated injection PMA group (0.5 µg/µl PMA, once a week for 4 weeks). The expression levels of p21, detected using double immunofluorescence staining with Iba‑1, and β‑galactosidase, via double immunohistochemical staining of Iba‑1, were examined in these three groups. The results indicated that a single injection of PMA did not change the microglial morphology and had no significant effects on the expression levels of TNF‑α and IL‑1β, compared with the control group (P>0.05). Following four repeated injections of PMA, the microglia in the substantia nigra presented with features of senescence, characterized by increased expression levels of β-galactosidase (P<0.001) and p21 (P<0.001), compared with the repeated injection control group. In conclusion, repeated intra-nigrostriatal treatment with PMA induced microglial senescence with increased expression levels of β-galactosidase and p21 in the substantia nigra of the rats.

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1	Kielian T: Microglia and chemokines in infectious diseases of the nervous system: Views and reviews. Front Biosci. 9:732–750. 2004. View Article : Google Scholar : PubMed/NCBI
2	Kanaan NM, Kordower JH and Collier TJ: Age and region-specific responses of microglia, but not astrocytes, suggest a role in selective vulnerability of dopamine neurons after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine exposure in monkeys. Glia. 56:1199–1214. 2008. View Article : Google Scholar : PubMed/NCBI
3	Overmyer M, Helisalmi S, Soininen H, Laakso M, Riekkinen P Sr and Alafuzoff I: Reactive microglia in aging and dementia: An immunohistochemical study of postmortem human brain tissue. Acta Neuropathol. 97:383–392. 1999. View Article : Google Scholar : PubMed/NCBI
4	Luo XG, Ding JQ and Chen SD: Microglia in the aging brain: Relevance to neurodegeneration. Mol Neurodegener. 5:122010. View Article : Google Scholar : PubMed/NCBI
5	Sawada M, Sawada H and Nagatsu T: Effects of aging on neuroprotective and neurotoxic properties of microglia in neuro-degenerative diseases. Neurodegener Dis. 5:254–256. 2008. View Article : Google Scholar
6	Conde JR and Streit WJ: Effect of aging on the microglial response to peripheral nerve injury. Neurobiol Aging. 27:1451–1461. 2006. View Article : Google Scholar
7	Streit WJ, Sammons NW, Kuhns AJ and Sparks DL: Dystrophic microglia in the aging human brain. Glia. 45:208–212. 2004. View Article : Google Scholar : PubMed/NCBI
8	Streit WJ, Miller KR, Lopes KO and Njie E: Microglial degeneration in the aging brain-bad news for neurons? Front Biosci. 13:3423–3438. 2008. View Article : Google Scholar : PubMed/NCBI
9	Olanow CW and Tatton WG: Etiology and pathogenesis of Parkinson's disease. Annu Rev Neurosci. 22:123–144. 1999. View Article : Google Scholar : PubMed/NCBI
10	Tha KK, Okuma Y, Miyazaki H, Murayama T, Uehara T, Hatakeyama R, Hayashi Y and Nomura Y: Changes in expressions of proinflammatory cytokines IL-1beta, TNF-alpha and IL-6 in the brain of senescence accelerated mouse (SAM) P8. Brain Res. 885:25–31. 2000. View Article : Google Scholar : PubMed/NCBI
11	Campisi J: The biology of replicative senescence. Eur J Cancer. 33:703–709. 1997. View Article : Google Scholar : PubMed/NCBI
12	Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D and Hwang ES: Senescence-associated beta-galactosidase is lysosomal beta-galactosidase. Aging Cell. 5:187–195. 2006. View Article : Google Scholar : PubMed/NCBI
13	Roninson IB: Oncogenic functions of tumour suppressor p21 (Waf1/Cip1/Sdi1): Association with cell senescence and tumour-promoting activities of stromal fibroblasts. Cancer Lett. 179:1–14. 2002. View Article : Google Scholar : PubMed/NCBI
14	Macip S, Igarashi M, Fang L, Chen A, Pan ZQ, Lee SW and Aaronson SA: Inhibition of p21-mediated ROS accumulation can rescue p21-induced senescence. EMBO J. 21:2180–2188. 2002. View Article : Google Scholar : PubMed/NCBI
15	Campisi J and d'Adda di Fagagna F: Cellular senescence: When bad things happen to good cells. Nat Rev Mol Cell Biol. 8:729–740. 2007. View Article : Google Scholar : PubMed/NCBI
16	Blumberg PM: Protein kinase C as the receptor for the phorbol ester tumor promoters: Sixth Rhoads memorial award lecture. Cancer Res. 48:1–8. 1988.PubMed/NCBI
17	Li YH, Bi HC, Huang L, Jin J, Zhong GP, Zhou XN and Huang M: Phorbol 12-myristate 13-acetate inhibits P-glycoprotein-mediated efflux of digoxin in MDCKII-MDR1 and Caco-2 cell monolayer models. Acta Pharmacologica Sin. 35:283–291. 2014. View Article : Google Scholar
18	O'neill AK, Gallegos LL, Justilien V, Garcia EL, Leitges M, Fields AP, Hall RA and Newton AC: Protein kinase Cα promotes cell migration through a PDZ-dependent interaction with its novel substrate discs large homolog 1 (DLG1). J Biol Chem. 286:43559–43568. 2011. View Article : Google Scholar : PubMed/NCBI
19	Mason SA, Cozzi SJ, Pierce CJ, Pavey SJ, Parsons PG and Boyle GM: The induction of senescence-like growth arrest by protein kinase C-activating diterpene esters in solid tumor cells. Invest New Drugs. 28:575–586. 2010. View Article : Google Scholar
20	Akakura S, Nochajski P, Gao L, Sotomayor P, Matsui S and Gelman IH: Rb-dependent cellular senescence, multinucleation and susceptibility to oncogenic transformation through PKC scaffolding by SSeCKS/AKAP12. Cell Cycle. 9:4656–4665. 2010. View Article : Google Scholar : PubMed/NCBI
21	Grealish S, Xie L, Kelly M and Dowd E: Unilateral axonal or terminal injection of 6-hydroxydopamine causes rapid-onset nigrostriatal degeneration and contralateral motor impairments in the rat. Brain Res Bull. 77:312–319. 2008. View Article : Google Scholar : PubMed/NCBI
22	Koziorowski D, Friedman A, Arosio P, Santambrogio P and Dziewulska D: ELISA reveals a difference in the structure of substantia nigra ferritin in Parkinson's disease and incidental lewy body compared to control. Parkinsonism Relat Disord. 13:214–218. 2007. View Article : Google Scholar : PubMed/NCBI
23	Itahana K, Zou Y, Itahana Y, Martinez JL, Beausejour C, Jacobs JJ, Van Lohuizen M, Band V, Campisi J and Dimri GP: Control of the replicative life span of human fibroblasts by p16 and the polycomb protein Bmi-1. Mol Cell Biol. 23:389–401. 2003. View Article : Google Scholar :
24	West MJ, Slomianka L and Gundersen HJ: Unbiased stereological estimation of the total number of neurons in thesubdivisions of the rat hippocampus using the optical fractionator. Anat Rec. 231:482–497. 1991. View Article : Google Scholar : PubMed/NCBI
25	Pan J, Wang G, Yang HQ, Hong Z, Xiao Q, Ren RJ, Zhou HY, Bai L and Chen SD: K252a prevents nigral dopaminergic cell death induced by 6-hydroxydopamine through inhibition of both mixed-lineage kinase 3/c-Jun NH2-terminal kinase 3 (JNK3) and apoptosis-inducing kinase 1/JNK3 signaling pathways. Mol Pharmacol. 72:1607–1618. 2007. View Article : Google Scholar : PubMed/NCBI
26	German DC and Manaye KF: Midbrain dopaminergic neurons (nuclei A8, A9 and A10): Three-dimensional reconstruction in the rat. J Comp Neurol. 331:297–309. 1993. View Article : Google Scholar : PubMed/NCBI
27	Streit WJ, Braak H, Xue QS and Bechmann I: Dystrophic (senescent) rather than activated microglial cells are associated with tau pathology and likely precede neurodegeneration in Alzheimer's disease. Acta Neuropathol. 118:475–485. 2009. View Article : Google Scholar : PubMed/NCBI
28	Luo XG and Chen SD: The changing phenotype of microglia from homeostasis to disease. Transl Neurodegener. 1:92012. View Article : Google Scholar : PubMed/NCBI
29	Kim YS and Joh TH: Microglia, major player in the brain inflammation: Their roles in the pathogenesis of Parkinson's disease. Exp Mol Med. 38:333–347. 2006. View Article : Google Scholar : PubMed/NCBI
30	Franceschi C, Bonafé M, Valensin S, Olivieri F, De Luca M, Ottaviani E and De Benedictis G: Inflamm-aging. An evolutionary perspective on immunosenescence. Ann N Y Acad Sci. 908:244–254. 2000. View Article : Google Scholar : PubMed/NCBI
31	Inamizu T, Chang MP and Makinodan T: Influence of age on the production and regulation of interleukin-1 in mice. Immunology. 55:447–455. 1985.PubMed/NCBI
32	Corsini E, Battaini F, Lucchi L, Marinovich M, Racchi M, Govoni S and Galli CL: A defective protein kinase C anchoring system underlying age–zassociated impairment in TNF-alpha production in rat macrophages. J Immunol. 163:3468–3473. 1999.PubMed/NCBI
33	Plackett TP, Boehmer ED, Faunce DE and Kovacs EJ: Aging and innate immune cells. J Leukoc Biol. 76:291–299. 2004. View Article : Google Scholar : PubMed/NCBI
34	Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I and Pereira-Smith O: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA. 92:9363–9367. 1995. View Article : Google Scholar : PubMed/NCBI
35	Yu HM, Zhao YM, Luo XG, Feng Y, Ren Y, Shang H, He ZY, Luo XM, Chen SD and Wang XY: Repeated lipopolysac-charide stimulation induces cellular senescence in BV2 cells. Neuroimmunomodulation. 19:131–136. 2012. View Article : Google Scholar
36	Petiti JP, De Paul AL, Gutiérrez S, Palmeri CM, Mukdsi JH and Torres AI: Activation of PKC epsilon induces lactotroph proliferation through ERK1/2 in response to phorbol ester. Mol Cell Endocrinol. 289:77–84. 2008. View Article : Google Scholar : PubMed/NCBI
37	Inoue S, Hao Z, Elia AJ, Cescon D, Zhou L, Silvester J, Snow B, Harris IS, Sasaki M, Li WY, et al: Mule/Huwe1/Arf-BP1 suppresses Ras-driven tumorigenesis by preventing c-Myc/Miz1-mediated down-regulation of p21 and p15. Genes Dev. 27:1101–1114. 2013. View Article : Google Scholar : PubMed/NCBI
38	Mowla S, Pinnock R, Leaner VD, Goding CR and Prince S: PMA-induced up-regulation of TBX3 is mediated by AP-1 and contributes to breast cancer cell migration. Biochem J. 433:145–153. 2011. View Article : Google Scholar
39	Serrano M, Lin AW, Mccurrach ME, Beach D and Lowe SW: Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell. 88:593–602. 1997. View Article : Google Scholar : PubMed/NCBI
40	John R, Chand V, Chakraborty S, Jaiswal N and Nag A: DNA damage induced activation of Cygb stabilizes p53 and mediates G1 arrest. DNA Repair (Amst). 2014:107–112. 2014. View Article : Google Scholar
41	Sarkisian CJ, Keister BA, Stairs DB, Boxer RB, Moody SE and Chodosh LA: Dose-dependent oncogene-induced senescence in vivo and its evasion during mammary tumorigenesis. Nat Cell Biol. 9:493–505. 2007. View Article : Google Scholar : PubMed/NCBI
42	Courtois-Cox S, Genther Williams SM, Reczek EE, Johnson BW, McGillicuddy LT, Johannessen CM, Hollstein PE, MacCollin M and Cichowski K: A negative feedback signaling network underlies oncogene-induced senescence. Cancer Cell. 10:459–472. 2006. View Article : Google Scholar : PubMed/NCBI
43	Chinta SJ, Lieu CA, Demaria M, Laberge RM, Campisi J and Andersen JK: Environmental stress, ageing and glial cell senescence: A novel mechanistic link to Parkinson's disease? J Intern Med. 273:429–436. 2013. View Article : Google Scholar : PubMed/NCBI
44	Kumar A, Chen SH, Kadiiska MB, Hong JS, Zielonka J, Kalyanaraman B and Mason RP: Inducible nitric oxide synthase is key to peroxynitrite-mediated, LPS-induced protein radical formation in murine microglial BV2 cells. Free Radical Bio Med. 73:51–59. 2014. View Article : Google Scholar
45	Collado M and Serrano M: Senescence in tumours: Evidence from mice and humans. Nat Rev Cancer. 10:51–57. 2010. View Article : Google Scholar
46	Parrinello S, Samper E, Krtolica A, Goldstein J, Melov S and Campisi J: Oxygen sensitivity severely limits the replicative lifespan of murine fibroblasts. Nat Cell Biol. 5:741–747. 2003. View Article : Google Scholar : PubMed/NCBI
47	Chen JH, Ozanne SE and Hales CN: Methods of cellular senescence induction using oxidative stress. Methods Mol Biol. 371:179–189. 2007. View Article : Google Scholar : PubMed/NCBI
48	Bitto A, Sell C, Crowe E, Lorenzini A, Malaguti M, Hrelia S and Torres C: Stress-induced senescence in human and rodent astrocytes. Exp Cell Res. 316:2961–2968. 2010. View Article : Google Scholar : PubMed/NCBI
49	Inoue Y, Matsuda T, Sugiyama KI, Izawa S and Kimura A: Genetic analysis of glutathione peroxidase in oxidative stress response of Saccharomyces cerevisiae. J Biol Chem. 274:27002–27009. 1999. View Article : Google Scholar : PubMed/NCBI
50	Kaneto H, Kajimoto Y, Fujitani Y, Matsuoka T, Sakamoto K, Matsuhisa M, Yamasaki Y and Hori M: Oxidative stress induces p21 expression in pancreatic islet cells: Possible implication in beta-cell dysfunction. Diabetologia. 42:1093–1097. 1999. View Article : Google Scholar : PubMed/NCBI
51	Croce CM: Oncogenes and cancer. N Engl J Med. 358:502–511. 2008. View Article : Google Scholar : PubMed/NCBI
52	Streit WJ and Xue QS: Human CNS immune senescence and neurodegeneration. Curr Opin Immunol. 29:93–96. 2014. View Article : Google Scholar : PubMed/NCBI
53	Lehmann AR and Carr AM: The ataxia-telangiectasia gene: A link between checkpoint controls, neurodegeneration and cancer. Trends Genet. 11:375–377. 1995. View Article : Google Scholar : PubMed/NCBI
54	Vanacore N, Spila-Alegiani S, Raschetti R and Meco G: Mortality cancer risk in parkinsonian patients: A population-based study. Neurology. 52:395–398. 1999. View Article : Google Scholar : PubMed/NCBI
55	Ong EL, Goldacre R and Goldacre M: Differential risks of cancer types in people with Parkinson's disease: A national record-linkage study. Eur J Cancer. 50:2456–2462. 2014. View Article : Google Scholar : PubMed/NCBI