Enhancing the efficacy of Hypericum perforatum in the treatment of an experimental model of multiple sclerosis using gold nanoparticles: an in vivo study

Document Type : Original Research Article

Authors

1 Immunology Research Center, Department of Immunology and Allergy, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Laboratory Sciences, Faculty of Paramedicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran. Immunology of Infectious Diseases Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

3 Department of Physiology, School of Medicine, Jiroft University of Medical Sciences, Jiroft, Iran

4 Department of Medical Immunology, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran

Abstract

Objective: Hypericum perforatum is a herbal medicine used in traditional medicine for the treatment of depression due to its antidepressant and anti-inflammatory activities. Therefore, we evaluated the therapeutic efficacy of H. perforatum extract (HPE) in combination with gold nanoparticles (HPE-GNP) against experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis.
Materials and Methods: EAE was induced in C57BL/6 mice with subcutaneous injection of MOG35-55 emulsified in complete Freund's adjuvant, and intraperitoneal pertussis toxin. Mice were treated with drugs in free (HPE) and nano-form (HPE-GNP) preparations. Splenocytes were isolated from all mice and the level of inflammatory and anti-inflammatory cytokines were evaluated by ELISA. The expression of T cells' transcription factors was also assessed using Real-Time PCR.
Results: Clinical score was reduced after HPE-GNP treatment. This change was associated with a decrease in the incidence and infiltration of inflammatory cells into the central nervous system. Additionally, treatment with HPE-GNP decreased the level of pro-inflammatory cytokines (IFN-γ, IL-17A and IL-6) and increased anti-inflammatory cytokines (TGF-β, IL-10 and IL-4). The real-time analysis revealed a decrease in the level of T-bet and ROR-γt  but an increase in FoxP3 and GATA3 expression.
Conclusion: The current study demonstrated that HPE-GNP could potentially reduce clinical and pathological complications of EAE, but laboratory data showed that HPE-GNP was significantly more effective than HPE in the treatment of EAE.

Keywords


Acharya S, Timilshina M, Jiang, L, Neupane
S, Choi DY, Park SW, Lee SY, Jeong BS,
Kim JA, Nam TG. 2018. Amelioration of
experimental autoimmune
encephalomyelitis and DSS induced
colitis by NTG-A-009 through the
inhibition of Th1 and Th17 cells
differentiation. Sci Rep, 8: 1-14.
Altan A, Aras MH, Damlar I, Gokce H, Ozcan
O, Alpaslan C. 2018. The effect of
Hypericum Perforatum on wound healing
of oral mucosa in diabetic rats. Eur Oral
Res, 52: 143-149.
Badeggi UM, Ismail E, Adeloye AO, Botha S,
Badmus JA, Marnewick JL, Cupido CN,
Hussein AA. 2020. Green synthesis of
HPE-GNP alleviates EAE
AJP, Vol. 12, No. 3, May-Jun 2022 335
gold nanoparticles capped with
procyanidins from Leucosidea sericea as
potential antidiabetic and antioxidant
agents. Biomolecules, 10: 1-24.
Bailly AL, Correard F, Popov A, Tselikov G,
Chaspoul F, Appay R, Al-Kattan A,
Kabashin AV, Braguer D, Esteve MA.
2019. In vivo evaluation of safety,
biodistribution and pharmacokinetics of
laser-synthesized gold nanoparticles. Sci
Rep, 9: 1-12.
Bettelli E, Sullivan B, Szabo SJ, Sobel RA,
Glimcher LH, Kuchroo VK. 2004. Loss
of T-bet, but not STAT1, prevents the
development of experimental autoimmune
encephalomyelitis. J Exp Med, 200: 79-
87.
Bonaterra GA, Schwendler A, Huther J,
Schwarzbach H, Schwarz A, Kolb C,
Abdel-Aziz H, Kinscherf R. 2017.
Neurotrophic, cytoprotective, and antiinflammatory effects of St. John's Wort
extract on differentiated mouse
hippocampal HT-22 neurons. Front
Pharmacol, 8: 1-13.
Brenn A, Grube M, Jedlitschky G, Fischer A,
Strohmeier B, Eiden M, Keller M,
Groschup MH, Vogelgesang S. 2014. St.
John's Wort reduces beta‐amyloid
accumulation in a double transgenic
Alzheimer's disease mouse model—role
of p‐glycoprotein. Brain Pathol, 24: 18-
24.
Bretscher P. 2019. On analyzing how the
Th1/Th2 phenotype of an immune
response is determined: classical
observations must not be ignored. Front
Immunol, 10: 1-7.
Cabrelle A, Dell’aica I, Melchiori L, Carraro
S, Brunetta E, Niero R, Scquizzato E,
D’intino G, Calzà L, Garbisa S. 2008.
Hyperforin down-regulates effector
function of activated T lymphocytes and
shows efficacy against Th1-triggered
CNS inflammatory-demyelinating
disease. J Leukoc Biol, 83: 212-219.
Capone A, Volpe E. 2020. Transcriptional
regulators of T Helper 17 Cell
differentiation in health and autoimmune
diseases. Front Immunol, 11: 1-8.
Dai H, Ciric B, Zhang GX, Rostami A. 2012.
Interleukin-10 plays a crucial role in
suppression of experimental autoimmune
encephalomyelitis by Bowman-Birk
inhibitor. J Neuroimmunol, 245: 1-7.
Danikowski KM, Jayaraman S, Prabhakar BS.
2017. Regulatory T cells in multiple
sclerosis and myasthenia gravis. J
Neuroinflammation, 14: 1-16.
Dauncey EA, Irving JTW, Allkin R. 2019. A
review of issues of nomenclature and
taxonomy of Hypericum perforatum L.
and Kew's Medicinal Plant Names
Services. J Pharm Pharmacol, 71: 4-14.
Dobson R, Giovannoni G. 2019. Multiple
sclerosis–a review. Eur J Neurol, 26: 27-
40.
Froushani SMA, Gouvarchin Galee HE,
Khamisabadi M, Lotfallahzade B. 2015.
Immunomudulatory effects of
hydroalcoholic extract of Hypericum
perforatum. Avicenna J Phytomed, 5: 62-
68.
Gajofatto A, Benedetti MD. 2015. Treatment
strategies for multiple sclerosis: when to
start, when to change, when to stop?
World J Clin Cases, 3: 545-555.
Gomez DR, Sánchez-Reus M, Iglesias I, Pozo
M, Garcia-Arencibia M, Fernández-Ruiz
J, Garcia-Garcia L, Delgado M, Benedi J.
2013. Neuroprotective properties of
standardized extracts of hypericum
perforatum on rotenone model of
Parkinson’s disease. CNS Neurol Disord
Drug Targets, 12: 665-679.
Haghmorad D, Salehipour Z, Nosratabadi R,
Rastin M, Kokhaei P, Mahmoudi MB,
Amini AA, Mahmoudi M. 2016.
Medium-dose estrogen ameliorates
experimental autoimmune
encephalomyelitis in ovariectomized
mice. J Immunotoxicol, 13: 885-896.
Huang N, Singh N, Yoon K, Loiacono CM,
Kohut ML, Birt DF. 2013. The immunoregulatory impact of orally-administered
Hypericum perforatum extract on Balb/C
mice inoculated with H1n1 influenza A
virus. PLoS One, 8: 1-20.
Komiyama Y, Nakae S, Matsuki T, Nambu A,
Ishigame H, Kakuta S, Sudo K, Iwakura
Y. 2006. IL-17 plays an important role in
the development of experimental
autoimmune encephalomyelitis. J
Immunol, 177: 566-573.
Kraus B, Wolff H, Heilmann J, Elstner EF.
2007a. Influence of Hypericum
perforatum extract and its single
compounds on amyloid-beta mediated
toxicity in microglial cells. Life Sci, 81:
884-894.
Mahmoudi et al.
AJP, Vol. 12, No. 3, May-Jun 2022 336
Kraus B, Wolff H, Heilmann J, Elstner EF.
2007b. Influence of Hypericum
perforatum extract and its single
compounds on amyloid-β mediated
toxicity in microglial cells. Life Sci, 81:
884-894.
Laidlaw BJ, Cui W, Amezquita RA, Gray SM,
Guan T, Lu Y, Kobayashi Y, Flavell RA,
Kleinstein SH, Craft J, Kaech SM. 2015.
Production of IL-10 by CD4(+)
regulatory T cells during the resolution of
infection promotes the maturation of
memory CD8(+) T cells. Nat Immunol,
16: 871-879.
Lassmann H, Bradl M. 2017. Multiple
sclerosis: experimental models and
reality. Acta Neuropathol, 133: 223-244.
Lovett-Racke AE, Yang Y, Racke, MK. 2011.
Th1 versus Th17: are T cell cytokines
relevant in multiple sclerosis? Biochim
Biophys Acta, 1812: 246-251.
Lu YH, Du CB, Liu JW, Hong W, Wei DZ.
2004. Neuroprotective effects of
Hypericum perforatum on trauma induced
by hydrogen peroxide in PC12 cells. Am
J Chin Med, 32: 397-405.
Martin R, Sospedra M, Rosito M, Engelhardt
B. 2016. Current multiple sclerosis
treatments have improved our
understanding of MS autoimmune
pathogenesis. Eur J Immunol, 46: 2078-
2090.
Mcginley AM, Edwards SC, Raverdeau M,
Mills KH. 2018. Th17 cells, γδ T cells
and their interplay in EAE and multiple
sclerosis. J Autoimmun, 87: 97-108.
Nosratabadi R, Rastin M, Sankian M,
Haghmorad D, Mahmoudi M. 2016a.
Hyperforin-loaded gold nanoparticle
alleviates experimental autoimmune
encephalomyelitis by suppressing Th1
and Th17 cells and upregulating
regulatory T cells. Nanomedicine, 12:
1961-1971.
Nosratabadi R, Rastin M, Sankian M,
Haghmorad D, Tabasi N, Zamani S,
Aghaee A, Salehipour Z, Mahmoudi M.
2016b. St. John’s Wort and its component
hyperforin alleviate experimental
autoimmune encephalomyelitis through
expansion of regulatory T-cells. J
immunotoxicol, 13: 364-374.
Oliveira AI, Pinho C, Sarmento B, Dias AC.
2016. Neuroprotective activity of
Hypericum perforatum and its major
components. Front Plant Sci, 7: 1-15.
Procaccini C, De Rosa V, Pucino V,
Formisano L, Matarese G. 2015. Animal
models of multiple sclerosis. Eur J
Pharmacol, 759: 182-191.
Salehipour Z, Haghmorad D, Sankian M,
Rastin M, Nosratabadi R, Soltan Dallal
MM, Tabasi N, Khazaee M, Nasiraii LR,
Mahmoudi M. 2017. Bifidobacterium
animalis in combination with human
origin of Lactobacillus plantarum
ameliorate neuroinflammation in
experimental model of multiple sclerosis
by altering CD4+ T cell subset balance.
Biomed Pharmacother, 95: 1535-1548.
Schepetkin IA, Özek G, Özek T, Kirpotina
LN, Khlebnikov AI, Quinn MT. 2020.
Chemical composition and
immunomodulatory activity of hypericum
perforatum essential oils. Biomolecules,
10: 1-20.
Segal BM. 2019. The diversity of
encephalitogenic CD4+ T cells in
multiple sclerosis and its animal models. J
Clin Med, 8: 1-13.
Sriwijitalai W, Wiwanitkit V. 2020. Herbs that
might be effective for the management of
COVID 19 A bioinformatics analysis on
anti-tyrosine kinase property. J Res Med
Sci, 2020: 1-2.
Teimuri-Mofrad R, Hadi R, Tahmasebi B,
Farhoudian S, Mehravar M, Nasiri R.
2017. Green synthesis of gold
nanoparticles using plant extract: Minireview. Nanochem Res, 2: 8-19.
Vecchia DD, Schamne MG, Ferro MM, Santos
AFCD, Latyki CL, Lara DVD, Ben J,
Moreira EL, Prediger RD, Miyoshi E.
2015. Effects of Hypericum perforatum
on turning behavior in an animal model of
Parkinson's disease. Braz J Pharm Sci, 51:
111-115.
Wagner CA, Roque PJ, Goverman JM. 2020.
Pathogenic T cell cytokines in multiple
sclerosis. J Exp Med, 217: 1-10.
Xiuying P, Jianping L, Ruofeng S, Liye Z,
Xuehong W, Yan L. 2012. Therapeutic
efficacy of Hypericum perforatum L.
extract for mice infected with an
influenza A