Protective effect of Artemisia absinthium on 6-hydroxydopamine-induced toxicity in SH-SY5Y cell line

Document Type : Original Research Article


1 Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Pharmacology and Medicinal plant Research Center, School of Medicine, Mashhad University of Medical Science, Mashhad, Iran.

3 Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Objective: Parkinson’s disease (PD) is a neurodegenerative disorder characterized by loss of dopaminergic neurons. Several experimental studies have shown neuroprotective and antioxidant effects for Artemisia absinthium. The present study was designed to assess the effect of A. absinthium on 6-hydroxydopamine (6-OHDA)-induced toxicity in SH-SY5Y cells.
Materials and Methods: SH-SY5Y cells were treated with ethanolic extract of A. absinthium for 24 hr and then, exposed to 6-OHDA (250 μM) for another 24 hr. MTT (3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyl tetrazolium bromide) assay was used for evaluation of cell viability. Moreover, the rate of apoptosis was measured using propidium iodide (PI) staining. The amount of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) was also measured using 2’, 7’–dichlorofluorescin diacetate (DCFDA) fluorometric method. Determination of glutathione (GSH) and superoxide dismutase (SOD) activity was done by colorimetric assay using DTNB [5, 5′-Dithiobis (2-nitrobenzoic acid)] and pyrogallol respectively.
Results: While 6-OHDA significantly increased ROS and apoptosis (p<0.001), the extract of A. absinthium significantly reduced ROS and cell apoptosis at concentrations ranging from 6.25 to 25 μg/mL (p<0.01 and p<0.001 respectively). Also, the extractsignificantly reduced MDA level in comparison with 6-OHDA (p<0.001). The GSH level and SOD activity were increased by the extract.
Conclusion: Findings of the current study showed that A. absinthium exerts it effect throughinhibiting oxidative stress parameters and it can be considered a promising candidate to be used in combination with the conventional medications for the treatment of neurodegenerative disorders, such as Parkinson's disease.


Amat N, Upur H, Blažeković B. 2010. In vivo
hepatoprotective activity of the aqueous
extract of Artemisia absinthium L. against
chemically and immunologically induced
liver injuries in mice. J Ethnopharmacol,
131: 478-484.
Aranda A, Sequedo L, Tolosa L, Quintas G,
Burello E, Castell JV, Gombau L. 2013.
Dichloro-dihydro-fluorescein diacetate
(DCFH-DA) assay: a quantitative method
for oxidative stress assessment of
nanoparticle-treated cells. Toxicol In Vitro,
Boncler M, Różalski M, Kraje wska U,
Podsędek A, Watala C. 2014. Comparison
of PrestoBlue and MTT assays of cellular
viability in the assessment of antiproliferative effects of plant extracts on
human endothelial cells. J Pharmacol Tox
Met, 69:9-16
Bora KS, Sharma A. 2010. Neuroprotective
effect of Artemisia absinthium L. on focal
ischemia and reperfusion-induced cerebral
injury. J Ethnopharmacol, 129:403-409
Bora KS, Sharma A. 2011. Evaluation of
antioxidant and free-radical scavenging
potential of Artemisia
absinthium. Pharm Biol, 49: 1216-1223
Canadanovic‐Brunet JM, Djilas SM, Cetkovic
GS, Tumbas VT. 2005. Free‐radical
scavenging activity of wormwood
(Artemisia absinthium L) extracts. J Sci
Food Agric, 85: 265-272
Chaturvedi RK, Shukla S, Seth K, Chauhan S,
Sinha C, Shukla Y, Agrawal AK. 2006.
Neuroprotective and neurorescue effect of
black tea extract in 6-hydroxydopaminelesioned rat model of Parkinson's disease.
Neurobiol Dis, 22: 421-434
Guarrera PM. 2005. Traditional phytotherapy
in Central Italy (Marche, Abruzzo, and
Latium). Fitoterapia, 76: 1-25
Haghdoost-Yazdi H, Sarookhani M, Faraj A,
Fraidouni N, Dargahi T, Yaghoubidoust
MH, Azhdari-Zarmehri H. Evaluation of the
association between blood homocysteine
concentration and the degree of behavioral
symptoms in the 6-hydroxydopamineinduced Parkinsonism in rat. 2014.
Pharmacol Biochem Behav, 124: 297-304
Hartmann A, Michel PP, Troadec JD, Mouatt‐
Prigent A, Faucheux BA, Ruberg M et al. Is
Bax a mitochondrial mediator in apoptotic
death of dopaminergic neurons in
Parkinson's disease?. 2001. J Neurochem,
76: 1785-1793.
Hosseini A, Mollazadeh H, Amiri MS,
Sadeghnia HR, Ghorbani A. Effects of a
standardized extract of Rheum
turkestanicum Janischew root on diabetic
changes in the kidney, liver and heart of
streptozotocin-induced diabetic rats. 2017.
Biomed Pharmaco, 86: 605-611.
Hujoel IA, Reilly NR, Rubio-Tapia A. Clinical
Features and Diagnosis. 2018. Gastroenterol
Clin, 48: 19-37
Ka H, Park H-J, Jung H-J, Choi J-W, Cho K-S,
Ha J, Lee KT. Cinnamaldehyde induces
apoptosis by ROS-mediated mitochondrial
permeability transition in human
promyelocytic leukemia HL-60 cells. 2003.
Cancer letters, 196: 143-152.
Keshavarz P, Gh BF, Mirhafez SR, Nematy M,
Azimi-Nezhad M, Afin SA, Esmaily H,
Pourali L, Hakak AM, Soukhtanloo M,
Mirteimouri M. Alterations in lipid profile,
zinc and copper levels and superoxide
dismutase activities in normal pregnancy
and preeclampsia. 2017. Am J Med Sci, 353:
Kharoubi O, Slimani M, Aoues A.
Neuroprotective effect of wormwood
against lead exposure. 2011. J Emerg
Trauma Shock, 4: 82-88.
Rashidi et al.
AJP, Vol. 11, No. 3, May-Jun 2021 246
Lee HG, Kim H, Oh WK, Yu KA, Choe YK,
Ahn JS, Kim DS, Kim SH, Dinarello CA,
Kim K, Yoon DY. 2004. tetramethoxy
hydroxyflavone p7f downregulates
inflammatory mediators via the inhibition of
nuclear factor κb. Ann NY Acad Sci, 1030:
Levites Y, Youdim MB, Maor G, Mandel S.
2002. Attenuation of 6-hydroxydopamine
(6-OHDA)-induced nuclear factor-kappaB
(NF-κB) activation and cell death by tea
extracts in neuronal cultures.
Biochem Pharmacol, 63: 21-29.
Li Y, Ohizumi Y. 2004. Search for constituents
with neurotrophic factor-potentiating
activity from the medicinal plants of
Paraguay and Thailand. Yakugaku Zasshi,
124: 417-424.
Lin CM, Lin YT, Lin RD, Huang WJ, Lee MH.
Neurocytoprotective effects of aliphatic
hydroxamates from Lovastatin, a secondary
metabolite from Monascus-fermented red
mold rice, in 6-hydroxydopamine (6-
OHDA)-treated nerve growth factor (NGF)-
differentiated PC12 cells. 2015. ACS Chem
Neurosci, 6: 716-724.
Mahmoudi M, Ebrahimzadeh MA, Ansaroudi
F, Nabavi SF, Nabavi SM. 2009.
Antidepressant and antioxidant activities of
Artemisia absinthium L. at flowering stage.
Afr J Biotechnol, 8: 7170-7175
Nikhat S, Ahmad S, Akhtar J, Jamil S.
Phytochemical and ethnopharmacological
perspective of Afsantin (Artemisia
absinthium Linn.). Ann Phytomed, 2013.
Pasban-Aliabadi H, Esmaeili-Mahani S,
Sheibani V, Abbasnejad M, Mehdizadeh A,
Yaghoobi MM. 2013. Inhibition of 6-
hydroxydopamine-induced PC12 cell
apoptosis by olive (Olea europaea L.) leaf
extract is performed by its main component
oleuropein. Rejuv Res, 16: 134-142.
Riccardi C, Nicoletti I. 2006. Analysis of
apoptosis by propidium iodide staining and
flow cytometry. Nat Protoc, 1:1458-1461
Ueda J, Kato J. 1980. Isolation and
identification of a senescence-promoting
substance from wormwood (Artemisia
absinthium L.). Plant Physiology, 66: 246-
Wake G, Court J, Pickering A, Lewis R,
Wilkins R, Perry E. 2000. CNS
acetylcholine receptor activity in European
medicinal plants traditionally used to
improve failing memory. J
Ethnopharmacol, 69: 105-114.
Zeng KW, Liao LX, Song XM, Lv HN, Song
FJ, Yu Q, Dong X, Jiang Y, Tu PF. 2015.
Caruifolin D from Artemisia absinthium L.
inhibits neuroinflammation via reactive
oxygen species-dependent c-jun N-terminal
kinase and protein kinase c/NF-κB signaling
pathways. Eur J Pharmacol, 767: 82-93.
Zhang Y, Dawson VL, Dawson TM. 2000.
Oxidative stress and genetics in the
pathogenesis of Parkinson's disease.
Neurobiol Dis, 7: 240-250.
Zhang ZJ, Cheang LC, Wang MW, Li GH, Chu
IK, Lin ZX, Lee SM. 2012. Ethanolic extract
of fructus Alpinia oxyphylla protects against
6-hydroxydopamine-induced damage of
PC12 cells in vitro and dopaminergic
neurons in zebrafish. Cell Mol Neurobiol,
32: 27-40
Zhang Z, Lv Z, Shao Y, Qiu Q, Zhang W, Duan
X, Li Y, Li C. 2017. Microsomal glutathione
transferase 1 attenuated ROS-induced lipid
peroxidation in Apostichopus japonicus.
Dev Comp Immunol, 73: 79-87