Evaluation of the in vivo and in vitro safety profile of Cuscuta epithymum ethanolic extract

Document Type : Original Research Article


1 Cellular and Molecular Research Center, Department of Pharmacology, Birjand University of Medical Sciences, Birjand, Iran

2 Student Research Committee, Birjand University of Medical Sciences, Birjand, Iran

3 Department of Pathology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran

4 Department of Physiology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran

5 Cellular and Molecular Research Center, Department of Biochemistry, Birjand University of Medical Sciences, Birjand, Iran

6 Cellular and Molecular Research Center,Department of Anatomical Sciences, Birjand University of Medical Sciences


Objective: Cuscuta epithymum (CE) is one of the most popular medicinal plants in the world. However, detailed information about its toxicity is not available. Hence, this study aimed to evaluate the safety profile of CE ethanolic extract in vitro and in vivo.
Materials and Methods: The extract's in vitro toxicity profile was investigated on normal fibroblast and cervical cancer cells by cytotoxicity test. In the next step, acute oral and intraperitoneal (i.p.) toxicity of the CE extract was evaluated in Wistar rats and BALB/c mice, respectively. Sub-acute oral toxicity was also examined by administering repeated oral doses of the CE extract (50, 200, and 500 mg/kg) to Wistar rats for 28 days.
Results: The CE extract exhibited a significant cytotoxicity on both normal (IC50 0.82 mg/ml, p<0.001) and cancer cells (IC50 1.42 mg/ml, p<0.001). Acute oral administration of a single dose of CE extract (175-5000 mg/kg) did not cause mortality; however, its i.p. administration caused mortality at doses greater than 75 mg/kg (i.p. LD50 154.8 mg/kg). In the sub-acute toxicity test, no significant effects in terms of weight change, organ weights, blood chemistry, or kidney pathology were observed. However, at 200 and 500 mg/kg doses, the CE extract significantly increased liver pathological scores compared to the control group (p<0.05 and p<0.01, respectively).
Conclusion: CE exhibited toxicities in i.p. acute and repeated oral dose administrations. It showed identical cytotoxicity against normal and cancer cells. This herb must be prescribed cautiously by traditional medicine practitioners.


Abedini MR, Wang PW, Huang YF, Cao M,
Chou CY, Shieh DB, Tsang BK. 2014. Cell
fate regulation by gelsolin in human
Abedini et al.
AJP, Vol. 11, No. 6, Nov-Dec 2021 654
gynecologic cancers. Proc Natl Acad Sci
USA, 111: 14442-14447.
Abutarbush SM. 2013. Alfalfa dodder (Cuscuta
campestris) toxicity in horses: clinical,
haematological and serum biochemical
findings. Vet Rec, 173: 95-98.
Amiri MS, Joharchi MR. 2013. Ethnobotanical
investigation of traditional medicinal plants
commercialized in the markets of Mashhad,
Iran. Avicenna J Phytomed, 3: 254-271.
Askari P, Namaei MH, Ghazvini K, Hosseini
M. 2021. In vitro and in vivo toxicity and
antibacterial efficacy of melittin against
clinical extensively drug-resistant bacteria.
BMC Pharmacol Toxicol, 22: 1-12.
Caporale F, Mateo-Martín J, Usman MF,
Smith-Hall C. 2020. Plant-Based
Sustainable Development—The Expansion
and Anatomy of the Medicinal Plant
Secondary Processing Sector in Nepal.
Sustainability, 12: 5575.
Chabra A, Monadi T, Azadbakht M,
Haerizadeh SI. 2019. Ethnopharmacology
of Cuscuta epithymum: A comprehensive
review on ethnobotany, phytochemistry,
pharmacology and toxicity. J
Ethnopharmacol, 231: 555-569.
Costea M, Tardif FJ. 2006. The biology of
Canadian weeds. 133. Cuscuta campestris
Yuncker, C. gronovii Willd. ex Schult., C.
umbrosa Beyr. ex Hook., C. epithymum (L.)
L. and C. epilinum Weihe. Can J Plant Sci,
86: 293-316.
Deniz U, Güneş H, Güneş F, Mammadov R.
2017. Cytotoxic activities of certain
medicinal plants on different cancer cell
lines. Turk J Pharm Sci, 14: 222-230.
Farzaei MH, Bayrami Z, Farzaei F, Aneva I,
Das SK, Patra JK, Das G, Abdollahi M.
2020. Poisoning by Medical Plants. Arch
Iran Med, 23: 117-127.
Firoozan J, Khodaie L, Mohammadi A, Fazljou
SM, Torbati M, Mohammadi Q, Mansoori
B, Bamdad S, Moghadam BB. 2020.
Dichloromethane EXTRACT of Cuscuta
epithymum Inhibits Triple-Negative Breast
Cancer Development Via Inducing
Apoptosis and Suppression of Migration. J
Biochem Tech, 11: 92-99.
Fiume MM, Bergfeld WF, Belsito DV, Hill RA,
Klaassen CD, Liebler DC, Marks Jr JG,
Shank RC, Slaga TJ, Snyder PW, Andersen
FA. 2014. Safety assessment of Vitis
vinifera (Grape)-derived ingredients as used
in cosmetics. Int J Toxicol, 33: 48S-83S.
Forouzanfar F, Vahedi MM, Aghaei A,
Rakhshandeh H. 2020. Hydroalcoholic
Extract of Cuscuta Epithymum Enhances
Pentobarbitalinduced Sleep: Possible
Involvement of GABAergic System. Curr
Drug Discov Technol, 17: 332-337.
Ganapaty S, Ramaiah M, Ramakrishna P,
Reddy DN. 2013. Scientific validation and
formulation of three Indian Folklore
medicinal plants. J Pharm Res, 6: 823-835.
Ghiravani Z, Hosseini M, Taheri MMH, Fard
MH, Abedini MR. 2016. Evaluation of
hypoglycemic and hypolipidemic effects of
internal septum of walnut fruit in alloxaninduced diabetic rats. Afr J Tradit
Complement Altern Med, 13: 94-100.
Hassanzadeh-Taheri M, Hassanpour-Fard M,
Doostabadi M, Moodi H, VazifeshenasDarmiyan K, Hosseini M. 2018a. Coadministration effects of aqueous extract of
turnip leaf and metformin in diabetic rats. J
Tradit Complement Med, 8: 178-183.
Hassanzadeh-Taheri M, Hosseini M,
Hassanpour-Fard M, Ghiravani Z,
Vazifeshenas-Darmiyan K, Yousefi S, Ezi
S. 2016. Effect of turnip leaf and root
extracts on renal function in diabetic rats.
Orient Pharm Exp Med, 16: 279-286.
Hassanzadeh-Taheri M, Hosseini M, Salimi M,
Moodi H, Dorranpour D. 2018b. Acute and
sub-acute oral toxicity evaluation of
Astragalus hamosus seedpod ethanolic
extract in Wistar rats. Pharm Sci, 24: 23-30.
Hassanzadeh‐Taheri M, Hassanzadeh‐Taheri
M, Jahani F, Hosseini M. 2018. Effects of
yoghurt butter oils on rat plasma lipids,
haematology and liver histology parameters
in a 150‐day study. Int J Dairy Technol, 71:
Hoshyar R, Jamali S, Fereidouni M, Abedini M.
2015. The cytotoxic activity of Ziziphus
Jujube on cervical cancer cells: In Vitro
study. Cell Mol Biol (Noisy-le-grand), 61:
Hoshyar R, Sebzari A, Balforoush M, Valavi
M, Hosseini M. 2019. The impact of Crocus
sativus stigma against methotrexate-induced
liver toxicity in rats. J Complement Integr
Med, 17: 1-9.
Jafarian A, Ghannadi A, Mohebi B. 2014.
Cytotoxic effects of chloroform and
hydroalcoholic extracts of aerial parts of
Cuscuta chinensis and Cuscuta epithymum
on Hela, HT29 and MDA-MB-468 tumor
cells. Res Pharm Sci, 9: 115-122.
Safety assessment of Cuscuta epithymum
AJP, Vol. 11, No. 6, Nov-Dec 2021 655
Jouad H, Haloui M, Rhiouani H, El Hilaly J,
Eddouks M. 2001. Ethnobotanical survey of
medicinal plants used for the treatment of
diabetes, cardiac and renal diseases in the
North centre region of Morocco (Fez–
Boulemane). J Ethnopharmacol, 77: 175-
Koca-Caliskan U, Yilmaz I, Taslidere A,
Yalcin FN, Aka C, Sekeroglu N. 2018.
Cuscuta arvensis Beyr “dodder”: in vivo
hepatoprotective effects against
acetaminophen-induced hepatotoxicity in
rats. J Med Food, 21: 625-631.
Kong Y, Chen D. 1996. Elucidation of Islamic
drugs in Hui Hui Yao Fang: a linguistic and
pharmaceutical approach. J
Ethnopharmacol, 54: 85-102.
Lipnick RL, Cotruvo JA, Hill RN, Bruce RD,
Stitzel KA, Walker AP, Chu I, Goddard M,
Segal L, Springer JA, Myers RC. 1995.
Comparison of the up-and-down,
conventional LD50, and fixed-dose acute
toxicity procedures. Food Chem Toxicol,
33: 223-231.
Lominadze Z, Kallwitz ER. 2018.
Misconception: You Can’t Have Liver
Disease With Normal Liver Chemistries.
Clin Liver Dis (Hoboken), 12: 96-99.
Marra F, Lotersztajn S. 2013. Pathophysiology
of NASH: perspectives for a targeted
treatment. Curr Pharm Des, 19: 5250-5269.
Mehrabani M, Modirian E, Ebrahimabadi A,
Vafazadeh J, Shahnavaz S, Heidari M. 2007.
Study of the effects of hydro-methanol
extracts of Lavandula vera DC. and Cuscuta
epithymum Murr. on the seizure induced by
pentylentetranzol in mice. J Kerman Univ
Med Sci, 14: 25-32.
Moodi H, Hosseini M, Abedini MR,
Hassanzadeh-Taheri M, HassanzadehTaheri M. 2020. Ethanolic extract of Iris
songarica rhizome attenuates methotrexateinduced liver and kidney damages in rats.
Avicenna J Phytomed, 10: 372-383.
Nagarajan D, Roy N, Kulkarni O, Nanajkar N,
Datey A, Ravichandran S, Thakur C,
Sandeep T, Aprameya IV, Sarma SP. 2019.
Ω76: A designed antimicrobial peptide to
combat carbapenem-and tigecyclineresistant Acinetobacter baumannii. Sci Adv,
5: eaax1946.
Ning ZH, Long S, Zhou YY, Peng ZY, Sun YN,
Chen SW, Su LM, Zhao YH. 2015. Effect of
exposure routes on the relationships of lethal
toxicity to rats from oral, intravenous,
intraperitoneal and intramuscular routes.
Regul Toxicol Pharmacol, 73: 613-619.
Parvizi M, Fadai F, Khodaei-Ardakani MR,
Amin G, Abdi L, Noroozi M, Ansari I. 2019.
Effect of Cuscuta epithymum acquainted
with risperidone on the improvement of
clinical symptoms and cognitive impairment
in patients with schizophrenia: A tripleblind randomized placebo-controlled trial.
Galen, 8: e1334.
Percie du Sert N, Hurst V, Ahluwalia A, Alam
S, Avey MT, Baker M, Browne WJ, Clark
A, Cuthill IC, Dirnagl U. 2020. The
ARRIVE guidelines 2.0: Updated guidelines
for reporting animal research. J Cereb Blood
Flow Metab, 40: 1769-1777.
Rispin A, Farrar D, Margosches E, Gupta K,
Stitzel K, Carr G, Greene M, Meyer W,
McCall D. 2002. Alternative methods for
the median lethal dose (LD50) test: the upand-down procedure for acute oral toxicity.
ILAR J, 43: 233-243.
Rout OP, Acharya R, Gupta R, Inchulkar SR,
Karbhal KS, Sahoo R. 2013. Management of
psychosomatic disorders through Ayurvedic
drugs-A critical review. World J Pharm
Pharm Sci, 2: 6507-6537.
Sellers RS, Morton D, Michael B, Roome N,
Johnson JK, Yano BL, Perry R, Schafer K.
2007. Society of Toxicologic Pathology
position paper: organ weight
recommendations for toxicology studies.
Toxicol Pathol, 35: 751-755.
Shah A, Bharati KA, Ahmad J, Sharma M.
2015. New ethnomedicinal claims from
Gujjar and Bakerwals tribes of Rajouri and
Poonch districts of Jammu and Kashmir,
India. J Ethnopharmacol, 166: 119-128.
Sharma P, Rani S, Ojha S, Sood S, Rana J.
2014. Indian herbal medicine as
hepatoprotective and hepatocurative: a
review of scientific evidence. Life Sci
Leaflets, 1: 61-115.
Sudam VS, Potnuri AG, Subhashini NP. 2017.
Syk–GTP RAC-1 mediated immunestimulatory effect of Cuscuta epithymum,
Ipomoea batata and Euphorbia hirta plant
extracts. Biomed Pharmacother, 96: 742-
Sutrisni NN, Soewandhi SN, Adnyana IK,
Sasongko LD. 2019. Acute and subchronic
(28-day) oral toxicity studies on the film
formulation of k-carrageenan and konjac
glucomannan for soft capsule application.
Sci Pharm, 87: 9.
Abedini et al.
AJP, Vol. 11, No. 6, Nov-Dec 2021 656
Wang Y, Ning ZH, Tai HW, Long S, Qin WC,
Su LM, Zhao YH. 2015. Relationship
between lethal toxicity in oral
administration and injection to mice: Effect
of exposure routes. Regul Toxicol
Pharmacol, 71: 205-212.
Winder C, Azzi R, Wagner D. 2005. The
development of the globally harmonized
system (GHS) of classification and labelling
of hazardous c