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Clinical Phytoscience
Published in: Clinical Phytoscience 1/2017

Open Access 01-12-2017 | Original contribution

Neuroprotective effects of Peltophorum pterocarpum leaf extract against hydrogen peroxide induced oxidative stress and cytotoxicity

Authors: Theanmalar Masilamani, Thavamanithevi Subramaniam, Norshariza Nordin, Rozita Rosli

Published in: Clinical Phytoscience | Issue 1/2017

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Abstract

Background

Peltophorum pterocarpum is a plant from the family of Leguminosae and a tropical tree found in South East Asia region. The leaves of the plant is rich in anti-oxidant polyphenols. Traditionally this plant was used to cure intestinal disorders and as a relief for sprain, bruise, muscular pain and sores and for pain during child birth. Since oxidative stress is a major contributing factor towards neurodegenerative diseases the leaves of this plant was explored for its neuroprotective properties.

Method

In this study, we report the neuroprotective properties of the ethanolic extract of Peltophorum pterocarpum leaf against oxidative stress induced cell death by H2O2, in an in vitro model of neuronally differentiated IMR32 neuroblastoma cells. Hydrogen peroxide was used as an inducer for oxidative stress. Cell viability was determined using MTT assay, apoptosis studies were carried out using Annexin V-FITC and Caspase 3 assays, mitochondrial membrane potential was examined using JC1 staining. The oxidative stress induced by H2O2 was determined by quantifying the intracellular reactive oxygen species production using DCF-DA staining. Mitogen activated protein kinase signaling pathway of the neuroprotective effect of PTE was also elucidated since oxidative stress activates this pathway signaling.

Results

Peltophorum pterocarpum leaf extract did not exhibit any cytotoxicity to differentiated IMR32 cells at the tested concentration of 7.8 μg/ml till 250 μg/ml. Pre-treatment of differentiated IMR32 with Peltophorum pterocarpum leaf extract prior to exposure to 300 μM hydrogen peroxide significantly ameliorated neuronal cell death and apoptosis in a dose-dependent manner. Hydrogen peroxide induced oxidative stress caused the reduction of mitochondrial membrane potential in differentiated IMR32 cells. Peltophorum pterocarpum leaf extract conferred neuroprotection to differentiated IMR32 by increasing the mitochondrial membrane potential in a dose-dependent manner which was significantly higher at 125 μg/ml and 250 μg/ml. PTE pre-treatment attenuated the increase of intracellular reactive oxygen species induced by hydrogen peroxide in a dose-dependent manner up till concentration of 125 μg/ml. Western blot analysis on mitogen activated protein kinases (MAPKs) revealed that neuroprotection of Peltophorum pterocarpum leaf extract against hydrogen peroxide induced cytotoxicity was achieved by complete inhibition of the activation of phospho-p-38 phosphorylation and attenuation of phospho-ERK1/2.

Conclusion

This study thus suggests that Peltophorum pterocarpum leaf extract has neuroprotective activity against oxidative-stress induced neuronal cell death.
Literature
1.
Uttara B, Singh AV, Zamboni P, Mahajan RT. Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol. 2009;7(1):65–74.CrossRefPubMedPubMedCentral
2.
Halliwell B, Gutteridge J. Cellular response to oxidative stress: adaptation, damage, repair, senescence and death. In: Free radical in biology and medicine. 4th ed. Oxford: Oxford University Press; 2007. p. 187–267.
3.
Yang F, Lim GP, Begum AN, Ubeda OJ, Simmons MR, Ambeguokar SS, Chen P, Kajed R, Glabe CG, Frautschy SA, Cole GM. Curcumin inhibits formation of amyloid beta oligomers and fibrils, binds plaques, and reduces amyloid in vivo. J Biol Chem. 2005;280(7):5892–901.CrossRefPubMed
4.
Ye J, Zhang Y. Curcumin protects against intracellular amyloid toxicity in rat primary neurons. Int J Clin Exp Med. 2012;5(1):44–9.PubMedPubMedCentral
5.
Damar U, Gersner R, Johnstone JT, Schachter S, Rotenberg A. Huperzine A as a neuroprotective and antiepileptic drug: a review of preclinical research. Expert Rev Neurother. 2016;16(6):671–80.CrossRefPubMed
6.
Shi C, Liu J, Wu F, Yew DT. Ginkgo biloba extract in Alzheimer’s disease: from action mechanisms to medical practice. Int J Mol Sci. 2010;11(1):107–23.CrossRefPubMedPubMedCentral
7.
Biswas K, Kumar A, Babaria BA, Prabhu K, Setty SR. Hepatoprotective effect of leaves of Peltophorum pterocarpum against paracetamol Induced acute liver damage in rats. J Basic Clin Pharm. 2009;1(1):10–5.PubMed
8.
Sridharamurthy NB, Ashok B, Yogananda R. Evaluation of Antioxidant and Acetyl Cholinesterase inhibitory activity of Peltophorum pterocarpum in Scopolamine treated rats. Int J Drug Dev Res. 2012;4(3):115–27.
9.
Manaharan T, Teng LL, Appleton D, Ming CH, Masilamani T, Palanisamy UD. Antioxidant and antiglycemic potential of Peltophorum pterocarpum plant parts. Food Chem. 2011;129(4):1355–61.CrossRef
10.
Borutaite V, Brown GC. Caspases are reversibly inactivated by hydrogen peroxide. FEBS Lett. 2001;500(3):114–8.CrossRefPubMed
11.
Polster BM, Fiskum G. Mitochondrial mechanisms of neural cell apoptosis. J Neurochem. 2004;90(6):1281–9.CrossRefPubMed
12.
Ono K, Han J. The p38 signal transduction pathway: activation and function. Cell Signal. 2000;12:1–13.CrossRefPubMed
13.
Strauss WL, Kemper RR, Jayakar P, Kong CF, Hersh LB, Hilt DC, Rabin M. Human choline acetyltransferase gene maps to region 10q11-q22.2 by in situ hybridization. Genomics. 1991;9(2):396–8.CrossRefPubMed
14.
Resende RR, Adhikari AA. Cholinergic receptor pathways involved in apoptosis, cell proliferation and neuronal differentiation. Cell Commun Signal. 2009;7:20.CrossRefPubMedPubMedCentral
15.
Jash SK, Singh RK, Majhi S, Sarkar A, Gorai D. Peltophorum pterocarpum: chemical and pharmacological aspects. Int J Pharm Sci Res. 2014;5(1):26–36.
16.
Whittermore ER, Loo DT, Watt JA, Cotman CW. Peroxide-induced cell death in primary neuronal culture. Neuroscience. 1995;67(4):921–32.CrossRef
17.
Gulden M, Jess A, Kammann J, Maser E& Seibert H. Cytotoxic potency of H2O2 in cell cultures: Impact of cell concentration and exposure time. Free Radic Biol Med. 2010;49(8):1298–305.CrossRefPubMed
18.
Vermes I, Haanen C, Nakken HS, Reutelingserger C. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995;184(95):39–51.CrossRefPubMed
19.
Shaykhalishahi H, Yazdanparast R, Ha HH, Chang YT. Inhibition of H2O2-induced neuroblastoma cell cytotoxicity by a triazine derivative, AA3E2. Eur J Pharmacol. 2009;622(1-3):1–6.CrossRefPubMed
20.
Wang XY, He PY, Du J, Zhang JZ. Quercetin in combating H2O2 induced early cell apoptosis and mitochondrial damage to normal human keratinocytes. Chin Med J. 2010;123(5):532–6.PubMed
21.
Jiang B, Liu JH, Bao Y, An LJ. Hydrogen peroxide-induced apoptosis in PC12 cells and the protective effect of puerarin. Cell Biol Int. 2003;27(12):1025–31.CrossRefPubMed
22.
Sanchana M, Flaskas J, Hargreaves AJ. In vitro biomarkers of developmental neurotoxicity. In: Gupta RC, editors. Reproductive and Developmental Toxicology. 1st ed. Elsevier; 2011: Chapter 19, 227–252.
23.
Sattayasai J, Chaonapan P, Arkaravichie T, Saimpornkul R, Junnu S, Charoensilp P, Samar J, Jantaravinid J, Masaratara P, Suktitipat B, Manissorn J, Thongboonkerd V, Neungtom N, Moongkarndi P. Protective effects of mangosteen extract on H2O2-induced cytotoxicity in SK-N-SH cells and scopolamine-induced memory impairment in mice. PLoS One. 2013;8(12), e85053: 1–13.
24.
Son YO, Jang YS, Heo JS, Chang WT, Choi KC Lee JC. Apoptosis inducing factor plays a critical role in caspase independent, pyknotic cell death in hydrogen peroxide exposed cells. Apoptosis. 2009;14:796–808.
25.
Hampton MB, Orrenius S. Dual regulation of caspase activity by hydrogen peroxide: Implications for apoptosis. FEBS Lett. 1997;414(3):552–6.CrossRefPubMed
26.
Kim DK, Cho ES, Um HD. Caspase-dependent and -independent events in apoptosis induced by hydrogen peroxide. Exp Cell Res. 2000;257(1):82–8.CrossRefPubMed
27.
Ly JD, Grubb D, Lawen A. The mitochondrial membrane potential (DYm) in apoptosis; an update. Apoptosis. 2003;8:115–28.CrossRefPubMed
28.
Armenta MM, Ruiz CN, Rebollar DJ, Martinez E, Gomex PY. Oxidative stress associated with neuronal apoptosis in experimental models of epilepsy. Oxidative Med Cell Longev. 2014;2014:1–12.CrossRef
29.
Halliwell B, Clement MV, Ramalingam J, Long LH. Hydrogen peroxide. Ubiquitous in cell culture and in vivo? IUBMB Life. 2000;50(4-5):251–7.CrossRefPubMed
30.
Babich H, Liebling EJ, Burger RF, Zuckerbraun HL, Schuck AG. Choice of DMEM, formulated with or without pyruvate, plays an important role in assessing the in vitro cytotoxicity of oxidants and prooxidant nutraceuticals. In Vitro Cell Dev Biol Anim. 2009;45(5-6):226–33.CrossRefPubMed
31.
Kelts JL, Cali JJ, Duellman SJ, Shultz J. Altered cytotoxicity of ROS-inducing compounds by sodium pyruvate in cell culture medium depends on the location of ROS generation. Spring. 2015;4:269.CrossRef
32.
Long LH, Clement MV, Halliwell B. Artifacts in cell culture: rapid generation of hydrogen peroxide on addition of (−)-epigallocatechin, (−)-epigallocatechin gallate, (+)-catechin, and quercetin to commonly used cell culture media. Biochem Biophys Res Commun. 2000;273(1):50–3.CrossRefPubMed
33.
Halliwell B. Are polyphenols antioxidants or pro-oxidants? What do we learn from cell culture and in vivo studies? Arch Biochem Biophys. 2008;476(2):107–12.CrossRefPubMed
34.
Long LH, Halliwell B. The effects of oxaloacetate on hydrogen peroxide generation from ascorbate and epigallocatechin gallate in cell culture media: Potential for altering cell metabolism. Biochem Biophys Res Commun. 2011;406(1):20–4.CrossRefPubMed
35.
Chen L, Liu L, Yin J, Luo Y, Huang S. Hydrogen peroxide-induced neuronal apoptosis is associated with inhibition of protein phosphatase 2A and 5, leading to activation of MAPK pathway. Int J Biochem Cell Biol. 2009;41:1284–95.CrossRefPubMed
36.
Li Z, Theus MH, Wei L. Role of ERK 1/2 signaling in neuronal differentiation of cultured embryonic stem cells. Develop Growth Differ. 2006;48(8):513–23.CrossRef
37.
Sun Y, Liu W, Liu T, Feng X, Yang N, Zhou HF. Signaling pathway of MAPK / ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct. 2015;9893(February):1–5.
38.
English JD, Sweatt JD. Activation of p42 Mitogen- activated Protein Kinase in Hippocampal Long Term Potentiation. J Biol Chem. 1996;271(October 4):24329–32.CrossRefPubMed
39.
Pearson G, Robinson F, Gibson TB, Xu BE, Karandikar M, Berman K, Cobb M. Mitogen-Activated Protein (MAP) Kinase Pathways: Regulation and Physiological Functions. Endocr Rev. 2001;22(2):153–83. doi:10.​1210/​er.​22.​2.​153.PubMed
40.
Sweatt JD. The neuronal MAP kinase cascade: a biochemical signal integration system subserving synaptic plasticity and memory. J Neurochem. 2001;76(1):1–10.CrossRefPubMed
41.
Marshall C. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell. 1995;80(2):179–85.CrossRefPubMed
42.
Glotin AL, Calipel A, Brossas JY, Faussat AM, Tréton J, Mascarelli F. Sustained versus transient ERK1/2 signaling underlies the anti- and proapoptotic effects of oxidative stress in human RPE cells. Investig Ophthalmol Vis Sci. 2006;47(10):4614–23.CrossRef
43.
44.
Vauzour D, Vafeiadou K, Rodriguez-Mateos A, Rendeiro C, Spencer JPE. The neuroprotective potential of flavonoids: a multiplicity of effects. Genes Nutr. 2008;3(3-4):115–26.CrossRefPubMedPubMedCentral
45.
46.
Takeda K, Ichiijo H. Neuronal p38 MAPK signalling: An emerging regulator of cell fate and function in the nervous system. Genes Cells. 2002;7(11):1099–111.CrossRefPubMed
47.
Crossthwaite AJ, Hasan S, William R. Hydrogen peroxide-mediated phosphorylation of ERK1/2, AKt/PKB and JNK in cortical neurones: Dependence on Ca2+ and PI3-kinase. J Neurochem. 2002;80(1):24–35.CrossRefPubMed
48.
Luo Y, DeFranco DB. Opposing roles for ERK1/2 in neuronal oxidative toxicity: Distinct mechanisms of ERK1/2 action at early versus late phases of oxidative stress. J Biol Chem. 2006;281:16436–42.CrossRefPubMed
49.
Odaka H, Numakawa T, Adachi N, Ooshima Y, Nakajima S, Katanuma Y, Inoue T, Kanugi H. Cabergoline, dopamine D2 receptor agonist, prevents neuronal cell death under oxidative stress via reducing excitotoxicity. PLoS One. 2014;9(6):1–11.
50.
Kwon SH, Kim JA, Hong SI, Jung YH, Kim HC, Lee SY, Jang CG. Loganin protects against hydrogen peroxide-induced apoptosis by inhibiting phosphorylation of JNK, p38, and ERK 1/2 MAPKs in SH-SY5Y cells. Neurochem Int. 2011;58(4):533–41.CrossRefPubMed
51.
Hu XL, Niu YX, Zhang Q, Tian X, Gao LY, Guo LP, Meng WH, Zhao QC. Neuroprotective effects of Kukoamine B against hydrogen peroxide-induced apoptosis and potential mechanisms in SH-SY5Y cells. Environ Toxicol Pharmacol. 2015;40(1):230–40.CrossRefPubMed
52.
Ruffels J, Griffin M, Dickenson JM. Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells: role of ERK1/2 in H2O2-induced cell death. Eur J Pharmacol. 2004;483(2-3):163–73.CrossRefPubMed
Metadata
Title
Neuroprotective effects of Peltophorum pterocarpum leaf extract against hydrogen peroxide induced oxidative stress and cytotoxicity
Authors
Theanmalar Masilamani
Thavamanithevi Subramaniam
Norshariza Nordin
Rozita Rosli
Publication date
01-12-2017
Publisher
Springer Berlin Heidelberg
Published in
Clinical Phytoscience / Issue 1/2017
Electronic ISSN: 2199-1197
DOI
https://doi.org/10.1186/s40816-017-0054-7

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