Artemisia sieberi: leave phytochemical screening, and biological activity with special emphasis on anticancer and anthelminthic efficacy

[Artemisia sieberi: triagem fitoquímica de folhas e atividade biológica com ênfase especial na eficácia anticâncer e anti-helmíntica]

ABSTRACT

Intestinal helminths are a significant cause of death every year. Many developing countries do not have access to conventional medications. Moreover, several anthelminthic drugs are associated with side effects and progressive increase in drug resistance. Currently, there is growing interest in discovering new antiparasitic agents from ethnomedicinal plants. Therefore, the present study aimed to investigate phytochemical screening, in vitro cytotoxicity, and anthelminthic activity ofArtemisia sieberileaves methanolic extract (ASLE). Using gas chromatography-mass spectra (GC-MS), the bioactive constituents of ASLE were detected. Additionally, different extract concentrations were tested for their anticancer activities when applied to colon cancer cell line HCT116 and liver cells Huh-7. ASLE was prepared and tested in vitro as an anthelmintic using earthwormEisenia fetida. Sixteen different bioactive constituents were found to be present in ASLE using (GC-MS). Moreover, ASLE showed significant cytotoxicity against cancer cells. The LC₅₀ of ASLE was obtained at 353.7±2.54μg/mL for the HCT116 and 467.9±1.97 for the Huh-7cell lines. Compared to Mebendazole, ASLE (200mg/mL) produced paralysis and earthworm death by 6.730±0.517 and 7.334±0.118 min, respectively, while, mebendazole (10 mg/mL) showed 13.91±0.007 and 18.2±0.980 min, respectively. In addition, histological examination revealed noticeable abnormalities in the tegument architecture of treated worms. Our results showed that ASLE has anticancer and anthelminthic activity which encourages the undertaking of numerous in vivo studies to discover an effective treatment.

Keywords:
Artemisia sieberi; Cytotoxicity; Anthelmintic; Eisenia fetida

RESUMO

As helmintoses intestinais são uma causa significativa de morte todos os anos. Muitos países em desenvolvimento não têm acesso a medicamentos convencionais. Além disso, vários medicamentos anti-helmínticos estão associados a efeitos colaterais e ao aumento progressivo da resistência aos medicamentos. Atualmente, há um interesse crescente na descoberta de novos agentes antiparasitários a partir de plantas etnomedicinais. Portanto, o presente estudo teve como objetivo investigar a triagem fitoquímica, a citotoxicidade in vitro e a atividade anti-helmíntica do extrato metanólico das folhas de Artemisia sieberi (ASLE). Usando a cromatografia gasosa e o espectro de massa (GC-MS), os constituintes bioativos do ASLE foram detectados. Além disso, diferentes concentrações do extrato foram testadas quanto às suas atividades anticancerígenas quando aplicadas à linha celular de câncer de cólon HCT116 e às células hepáticas Huh-7. O ASLE foi preparado e testado in vitro como anti-helmíntico usando a minhoca Eisenia fetida. Verificou-se que dezesseis constituintes bioativos diferentes estavam presentes no ASLE usando (GC-MS). Além disso, o ASLE apresentou citotoxicidade significativa contra células cancerígenas. A LC50 do ASLE foi obtida em 353,7±2,54μg/mL para as linhas celulares HCT116 e 467,9±1,97 para as linhas celulares Huh-7. Em comparação com o mebendazol, o ASLE (200 mg/mL) produziu paralisia e morte da minhoca em 6,730±0,517 e 7,334±0,118 minutos, respectivamente, enquanto o mebendazol (10 mg/mL) apresentou 13,91±0,007 e 18,2±0,980 minutos, respectivamente. Além disso, o exame histológico revelou anormalidades notáveis na arquitetura do tegumento dos vermes tratados. Nossos resultados mostraram que a ASLE tem atividade anticâncer e anti-helmíntica, o que incentiva a realização de vários estudos in vivo para descobrir um tratamento eficaz.

Palavras-chave:
Artemisia sieberi; Citotoxicidade; Anti-helmíntico; Eisenia fetida

INTRODUCTION

Gastrointestinal helminths represent a main threat to both human health and the well-being of small ruminants worldwide (Stepek et al., 2004). They constitute a considerable economic challenge in ruminant farming systems and cause adverse health outcomes due to factors such as anemia, mortality, weight reduction, and decreased milk and meat output (Lira et al., 2008; Githiori et al., 2003).

It was estimated that over 2 billion individuals suffer from chronic infections caused by one or more gastrointestinal helminth parasites and approximately 300 million people have severe morbidity, of which 10,000-135,000 deaths occur per year. These parasites are widely distributed in tropical and subtropical regions, especially among communities with poverty, poor hygiene, and in developing nations (The World…, 2005; Husen et al., 2022).

Numerous drugs are available for treating a broad range of helminth infections, such as albendazole, mebendazole, and nitazoxanide. However, the effectiveness of these medications varies. The cure rate with nitazoxanide remained at 84% even after three treatments. Cure rates as low as 61% (400 mg) and 67% (800 mg) for albendazole, and 19% (single) and 45% (repeated) for mebendazole have been documented (Silva et al., 1997; Diaz et al., 2003 and Krepel et al., 1993). Additionally, signs of drug resistance development by various parasites have emerged in recent decades (Curico et al., 2022; Fissiha and Kinde, 2021). Furthermore, there is an increased risk of residues in the meat and milk of animals, as well as in the environment. (Emery et al., 2016; Torres-Acosta et al., 2012).

Colon cancer ranks as the second leading cause of cancer-related death globally. In 2020, it was estimated that over 1.9 million new cases of colorectal cancer and more than 930,000 deaths due to colorectal cancer occurred worldwide. By 2040, the burden of colorectal cancer is projected to surge to 3.2 million new cases annually, representing a 63% increase, with deaths reaching 1.6 million per year, signifying a 73% increase. Meanwhile, liver cancer stood as the sixth most commonly diagnosed cancer and the third leading cause of cancer-related death worldwide (Morgan et al., 2023; Sung et al., 2021). In that year, approximately 830,180 deaths were attributed to liver cancer internationally, accounting for nearly 8.3% of total cancer deaths. However, there are no extremely effective drugs to treat most cancers.

As a result, there exists a pressing necessity for alternative control measures to address this challenge (Boonmasawai et al., 2013). Medicinal plants utilized in traditional folk medicine can thus provide a source of cost-effective and efficient anthelmintic and anticancer agents.

Numerous species of Artemisia are prevalent across the Kingdom of Saudi Arabia. Among them, A. sieberi is a well-known species of the Asteraceae family that is widely distributed and renowned for its medicinal properties. it primarily inhabits the Northern and Central regions, notably in scattered populations found in Wadis (Badr et al., 2012; Al Otibi and Rizwana, 2019; Arbi et al., 2017). It was recognized more than 2000 years ago (Padosch et al., 2006). It is used widely in folk medicine as an anthelminthic and as an antidote to mushroom poisoning. Numerous studies have explored the advantageous properties of A. sieberi essential oil, including its impact on skin development (Kaboutari et al., 2015), antimicrobial (Irshaid et al., 2010), insecticidal (Negahban et al., 2006), nematocidal (Ardakani and Parhizkar, 2012), anti-malarial (Nahrevanian et al., 2012). Furthermore, research has documented the anticancer properties of A. sieberi species found in Saudi Arabia (Nasr et al., 2020). Additionally, A. sieberi serves as a feed supplement (Khalaji et al., 2011) and has been used in the treatment of diabetes according to traditional beliefs. However, its activity has been regarded to the presence of diverse secondary metabolites including phenolic, flavonoid, and a sesquiterpene lactone. Thus, this genusis is attracting attention from researchers all over the world.

There has not been much report in literature of ASLE phytochemical constituents, activity, and no study explored its anthelminthic efficiency. Therefore, this research aimed to screen the phytochemical constituent of ASLE from Saudi Arabia by GC-MS method. Investigation of its biological activity including assay for in vitro anticancer activity against colon cancer cell line HCT116 and liver cells Huh-7, and anthelminthic efficiency using earthworm E. fetida.

MATERIALS AND METHODS

In Al Badiya - Tabuk, Saudi Arabia, leaves of A. sieberi were gathered, and a taxonomist from the Botany and Microbiology Department at King Saud University, Riyadh, Saudi Arabia, authenticated the plant material in the herbarium. The methanolic extract of A. sieberi leaves (70% methanol) was prepared following the method outlined by Manikandan et al. (2008), with modifications as described below: The air-dried leaves of A. sieberi were pulverized into a powder using an electric blender (Senses, MG-503T, Korea). The resulting dried powder (100 g) was macerated in 70% methanol for 24 hours at 4 ºC, followed by percolation 5-7 times until complete extraction. After filtration, ethanol was separated from the extract using a vacuum evaporator set at 50 °C and low pressure. The crude extract was then lyophilized and stored at -20 °C until further use.

The phytochemical investigation of ASLE has been conducted using GC-MS equipment (Thermo Scientific Co.), specifically the Thermo GC-TRACE ultra ver.: 5.0 and Thermo MS DSQ II (Kanthal et al., 2014). Experimental conditions for the GC-MS system have been standardized as follows: TR 5-MS capillary standard non-polar column with dimensions of 30 meters in length, 0.25 mm in internal diameter, and a film thickness of 0.25 μm. The flow rate of the mobile phase (carrier gas: He) was set at 1.0 ml/min. In the gas chromatography segment, the temperature program (oven temperature) started at 40°C and was gradually increased to 250°C at a rate of 5°C/min. The injection volume was 1 μl. Samples dissolved in chloroform have been subjected to a mass range of 50-650 m/z, and the results have been analyzed using the Wiley Spectral library search program.

HCT116 (colon) cancer were procured from American Type Culture Collection (ATCC®, Manassas, VA, USA) and Huh-7 (liver) cancer cell lines were procured from Cell Line Services (CLS; GmBH, Germany). They were routinely cultivated in DMEM medium (Gibco, USA) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, USA). In an incubator with a humidified environment of 5% CO2, the cells were incubated at 37°C.

HCT116 (colon) cancer and Huh-7 (liver) cells were plated in 96 well plates (10⁴/well) and incubated in CO₂ incubator at 37^oC for 24 hours. The cells were then treated with different concentrations of plant extracts including (0, 50, 100, 250, and 500 µg/ml). Plant extract or doxorubicin (positive control) were added and the cells were incubated for a further 48 hours. Next, 10 μl of MTT (5 mg/ml in PBS) was added to each well. After a further 4h incubation at 37°C, the MTT was removed and the purple formazan product dissolved in acidified isopropanol and absorbance was measured at 540 nm on a microplate reader (BioTek, USA). Results were expressed as a percentage of cell viability with respect to untreated control cells (as 100%). The IC₅₀ values were calculated from dose-response curve using OriginPro software.

From agricultural grounds, a total of 25 earthworms, E. fetida, were gathered. In each Petri dish, five almost equal-sized worms were put. As a positive control, albendazole (10 mg/ml) was positive, and distilled water was used as a negative control. With concentrations of 50, 100, and 200 mg/ml, the ASLE extract was prepared in distilled water. When no movement was observed except when shaken vigorously, the time for paralysis was recorded. while the death time was recorded when did not show any movement by vigorous shaking nor when dipped in warm water (50°C) for the worms (Parida et al., 2010).

The E. fetida was cut up into small pieces, fixed in 10% buffered neutral formalin, and then prepared for paraffin embedding. Hematoxylin and eosin (HE) were used to stain thin slices (4 μm) cut using a rotatory microtome (Drury and Wallington, 1973). A digital camera (DP 73) mounted on the microscope was used to take pictures while components were being examined under light microscopy (Olympus BX61, Tokyo, Japan) at a magnification of x400. Measurements of worms' cuticular thickness were made and shown in micrometers (m) using ImageJ 1.53e software.

One-way analysis of variance (ANOVA) was used to examine the data in SigmaPlot® version 11.0 (Systat Software, Inc., Chicago, IL, USA). Differences between groups were considered significant at a p-value ≤ 0.01.

RESULT

A sample of ASLE has been subjected to GC-MS equipment to identify its bioactive ingredient. The analysis revealed 16 major peaks and the bioactive constituents corresponding to the peaks were identified as follows p-Cymene, o-Cymene, 2(3H)-Furanone, 5-ethenyldihydro-5-methyl, 1,5,7-Octatrien-3-ol, 3,7-dimethyl, 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2,4-Cycloheptadien-1-one, 2,6,6-trimethyl, 4a(2H)-Naphthalenecarboxylic acid, octahydro, Palmitoyl chloride, Xanthoxylin, n-Hexadecanoic acid, Nitrosine, Caryophyllene oxide, Santamarine, 1,4-Butanediol, 2,3-bis[(4-hydroxy-3-methoxyphenyl)methyl], 4H-1-Benzopyran-4-one, 5,7-dihydroxy-2-(3,4,5-trimethoxyphenyl, Eupatorin (Fig 1, Table 1). The GC‒MS of the methanol extracts in our study showed the presence of Cymene which is related to monocyclic monoterpenes and has been reported earlier from Artemisia monosperma extract (Saleh et al., 2024). Sesquiterpenes lactone, polyphenols, and terpenoid were also present in the methanol extracts (Al Otibi and Rizwana, 2019)

According to the MTT assay results, ASLE exhibited a concentration-dependent decrease in cell viability. At concentrations of 500μg/mL, it demonstrated toxicity against 53% of Huh-7 cells (Fig 2). Furthermore, ASLE was found to be safe for normal cells at concentrations up to 400μg/mL. ASLE induced cytotoxic effects on the HCT 116 cell line, with a high concentration of 500 μg/ml resulting in cell death rates of 60%. IC₅₀ indicates the dose of ASLE, inducing 50% Huh-7 (467.9±1.97µg/mL) and HCT (353.7±2.54µg/mL) cancer cell growth inhibition (Table 2).

Figure 1
GC-MS of the methanolic extract of A. sieberi showing 16 peaks with retention times ranging from 5.45 min to 31.11 min.

Table 1
The phytochemical composition of A. sieberi was determined by using GC-MS

No	tR (min)	Proposed compound	MW	Formula
1	5.45	p-Cymene	134	C10H14
2	5.51	o-Cymene	134	C10H14
3	5.74	2(3H)-Furanone, 5-ethenyldihydro-5-methyl-	126	C7H10O2
4	6.84	1,5,7-Octatrien-3-ol, 3,7-dimethyl-	152	C10H16O
5	7.49	4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-	144	C6H8O4
6	10.16	2,4-Cycloheptadien-1-one, 2,6,6-trimethyl- terpene	150	C10H14O
7	12.87	4a(2H)-Naphthalenecarboxylic acid, octahydro-	182	C11H18O2
8	14.73	Palmitoyl chloride	274	C16H31ClO
9	14.79	Xanthoxylin	196	C10H12O4
10	17.91	n-Hexadecanoic acid	256	C16H32O2
11	21.88	Nitrosine	308	C17H24O5
12	22.05	Caryophyllene oxide	220	C15H24O
13	22.31	Santamarine	248	C15H20O3
14	26.23	1,4-Butanediol, 2,3-bis[(4-hydroxy-3-methoxyphenyl)methyl]	362	C20H26O6
15	29.29	4H-1-Benzopyran-4-one, 5,7-dihydroxy-2-(3,4,5-trimethoxyphenyl)-	344	C18H16O7
16	31.11	Eupatorin polymethoxy flavone	344	C18H16O7

Figure 2
In vitro the cytotoxicity (MTT) assay results for the tested ASLE at various concentrations (µg/mL) against colon cancer cell line HCT116 and liver cells Huh-7 following 48 hours of incubation.

Table 2
In vitro the cytotoxicity (MTT) assay results for IC₅₀ (µg/ml) ASLE against colon cancer cell line HCT116 and liver cells Huh-7

	Cell lines and IC50 (µg/ml)
Cell lines	HCT	Huh-7
ASLE	353.7 ± 2.54	467.9 ± 1.97
Doxorubicin	2.7 ± 0.05

It was established that ASLE is antihelmintic to E. fetida, where ASLE 200 mg/mL is the most efficient dose, showed the time to death and paralysis was 7.334 ± 0.118 and 6.730 ± 0.517 min, respectively. While mebendazole showed less effect 13.91 ± 0.373 and 18.2 ± 0.980 min for paralysis and death time, respectively (Table 3). In contrast to the control group, where there are no changes to the uppermost layer of the cuticle, the plant extracts cause the upper layer of the cuticle to be destroyed, reducing the segment length of the worms (Fig 3).

Table 3
In vitro anthelmintic activity of ASLE

Test samples	Concentration (mg/ml)	Time is taken for paralysis (min.)	Time is taken for death (min.)
Control (H2O)	--	--	--
ASLE	50 mg/ml	37.164 ± 2.894 *#	37.936 ± 3.255 *#
	100 mg/ml	22.264 ± 0.724 *#	22.978 ± 0.749 *#
	200 mg/ml	6.730 ± 0.517 *#	7.334 ± 0.118 *#
Mebendazole	10 mg/ml	13.91 ± 0.373 *	18.2 ± 0.980 *

• Values are mean ± SD. All superscripts indicate significance at p ≤ 0.05, ^* compared to untreated (H₂O), ^# compared to mebendazole.

Figure 3
Histological changes in the cuticle of E. fetida with diverse treatments. (A) worms in distilled water (H₂O) (control). (B) worms in ASLE (200 mg/mL). (C) worms in mebendazole. Scale bar = 25 μm.

DISCUSSION

The emergence of resistant strains, the detection of anthelmintic drug residues in animal products, and concerns regarding the toxicity of synthetic drugs have sparked renewed interest in the utilization of natural products (Asase et al., 2005). GC-MS analysis of the leaf extract from the plant A. sieberi unveiled numerous bioactive compounds associated with plant secondary metabolites, including p-Cymene that linked to monocyclic monoterpenes, sesquiterpenes, flavonoids, and phenolic compounds. These compounds exhibit anti-cancer, anti-inflammatory, antioxidant, and anti-parasitic activities (Alwahibi et al., 2016; Salih et al., 2023). However, The variability in phytochemical compounds among Artemisia species may stem from the diverse physiological and biochemical responses of different species to environmental conditions. It has been documented that various morphogenetic, genetic, and environmental factors can influence the biosynthesis and accumulation of bioactive compounds (Škrovánková et al., 2012; Yang et al., 2018).

In the present study, ASLE has been evaluated for its in vitro anthelmintic activity using earthworms. E. fetida served as a suitable model, exploiting the physiological similarity between earthworms and several gastrointestinal parasites (Cáceres et al., 2017; Abu Hawsah et al., 2023). Moreover, in vitro test is favored over in vivo methods due to their cost-effectiveness, simplicity, and rapid results turnover (Jesús-Martínez et al., 2018).

It is recognized that changes in the worms' tegument are among the parameters indicating anthelmintic activity (Tansatit et al., 2012). The current study found that ASLE had greater anthelmintic efficacy against earthworms than mebendazole, which resulted in changes in tegument integrity as well as severe morphological damage such as tegument peeling and destruction as observed by microscopic examination of histological sections. The results of our current study are consistent with (Beshay, 2018) who demonstrated that a crude aqueous extract of Artemisia absinthium has therapeutic benefits on Hymenolepis. nana. In a dose-dependent manner, the data demonstrated worm paralysis, death, and ultrastructural abnormalities such as tegumental damage, lipid buildup, and destruction of the nephridial canal and intrauterine eggs. In addition, Artemisia absinthium treated mice had significantly lower Eggs Per Gram (EPG) and worm burden. Beshay (2018). In another in vitro study, crude extracts of Artemisia herba-alba showed anthelmintic activity against Haemonchus contortu (Ahmed et al., 2020). Additionally, this finding aligns with a previously documented activity of a plant within the same genus, Artemisia monosperma, which notably impacted the motility, viability, and tegument abnormalities of Eisenia fetida in vitro )Saleh et al., 2024(.

The effectiveness of ASLE is attributed to the presence of various bioactive phytochemical components, sesquiterpene lactones and flavonoids, which could potentially possess anthelmintic properties (El-Maggar, 2012; Kerboeuf et al., 2008). Also, artemisinins that are sesquiterpene lactones have a toxic effect on E. granulosus in vitro as well as on Trichinella spiralis (Abou Rayia et al., 2017; Lü et al., 2014). This activity is attributed to the cleavage of the endoperoxide bridge present in their chemical structure. Upon activation by heme released during hemoglobin digestion or by free ferrous iron, they release carbon-centered free radicals or cytotoxic reactive oxygen species, which subsequently cause damage to the parasites (Ho et al., 2014; Loo et al., 2017; Miller and Su, 2011). In addition, flavonoids have been shown to induce protein denaturation in worm tissue, leading to the death of the worms (Goodman, 1960).

Recently, cancers are becoming more resistant to current drugs. So there is a continuing need for new therapies to be developed. In recent years, there has been a growing interest in Artemisia-derived products as potential candidates for inhibiting the growth of cancerous cells (Sagar et al., 2006). The methanol extracts of A. sieberi in this study showed exceptional cytotoxic efficacy against all examined cancer cell lines. This may be a result of the presence of artemisinin and other sesquiterpene lactones, which have previously been documented for this species (Arab et al., 2006; Abbas et al., 2017).

In vitro and in vivo tests on Artemisinin which is a sesquiterpene lactone revealed that it had cytotoxic properties capable of restraining the growth of various cancer cell lines, including those associated with breast, prostate, colon, and liver cancers (Efferth, 2006; Hou et al., 2008). Furthermore, some studies indicate that Artemisinin and its bioactive derivatives can impede angiogenesis, a crucial process in tumor growth and metastasis (Crespo-Ortiz and Wei, 2012). Additionally, artemisinin was discovered to be an effective antitumor, anti-leishmanial, antifungal, and antibacterial agent (Appalasamy et al., 2014).

CONCLUSION

These results offer scientific substantiation for the medicinal properties of A. sieberi in traditional medicine, underscoring the importance of exploring alternatives for anthelmintic and anticancer agents, particularly those derived from plant sources. ASLE showed promise as valuable resources for potential medicinal and industrial compounds. GC-MS analysis of the leaf extract from the Saudi plant A. sieberi revealed numerous bioactive compounds associated with plant secondary metabolites, including p-Cymene related to monocyclic monoterpenes, sesquiterpenes lactones, flavonoids, and phenolic compounds, which exhibit anti-cancer and anthelmintic activity.

ACKNOWLEDGMENTS

This work was supported by the Researchers Supporting Project (RSP2024R3) at King Saud University (Riyadh, Saudi Arabia).

REFERENCE

• ABBAS, Z.K.; SAGGU, S.; REHMAN, H. et al. Ecological variations and role of heat shock protein in Artemisia judaica L. in response to temperature regimes of Tabuk, Saudi Arabia. Saudi J. Biol. Sci., v.24, p.1268-1273, 2017.
• ABOU RAYIA, D.M.; SAAD, A.E.; ASHOUR, D.S.; OREIBY, R.M. Implication of artemisinin nematocidal activity on experimental trichinellosis: in vitro and in vivo studies. Parasitol. Int., v.66, v.56-63, 2017.
• ABU HAWSAH, M.; AL-OTAIBI, T.; ALOJAYRI, G. et al. In vitro studies for the antiparasitic activities of Azadirachta indica extract. Food Sci. Technol., v.43, p.e117122, 2023.
• AHMED, A.H.; EJO, M.; FEYERA, T. et al. In vitro anthelmintic activity of crude extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus. J. Parasitol. Res., v.2020, p.4950196, 2020.
• AL OTIBI, F.; RIZWANA, H. Chemical composition, FTIR studies, morphological alterations, and antifungal activity of leaf extracts of Artemisia sieberi from Saudi Arabia. Int. J. Agric., v.21, p.1241-1248, 2019.
• ALWAHIBI, L.H.; ABDEL-MAGEED, W.M.; ABDELKADER, M. et al. Sesquiterpene lactones and flavonoids from Artemisia sieberi. Int. J. Pharm. Phytochem. Res., v.8, p.639-644, 2016.
• APPALASAMY, S.; LO, K.Y.; CH'NG, S.J. et al. Antimicrobial activity of artemisinin and precursor derived from in vitro plantlets of Artemisia annua L. Biomed. Res. Int., v.2014, p.215872, 2014.
• ARAB, H.; RAHBARI, S.; RASSOULI, A. et al. Determination of artemisinin in Artemisia sieberi and anticoccidial effects of the plant extract in broiler chickens. Trop. Anim. Health Prod., v.38, p.497-503, 2006.
• ARBI, A.G.; AL-GHAMDI, F.; OSMAN, A. The genus Artemisia L. in the northern region of Saudi Arabia: essential oil variability and antibacterial activities. Nat. Prod. Res., v.31, p.598-603, 2017.
• ARDAKANI, A.S.; PARHIZKAR, S. Inhibitory effects of Teucrium polium L., Artemisia sieberi Besser. and Achillea wilhelmsii C. Koch on Meloidogyne incognita (Kofoid and White) Chitwood (in vitro and under greenhouse conditions). Int. J. Med. Arom. Plants, v.2, p.596-602, 2012.
• ASASE, A.; OTENG-YEBOAH, A.A.; ODAMTTEN, G.T.; SIMMONDS, M.S. Ethnobotanical study of some Ghanaian anti-malarial plants. J. Ethnopharmacol., v.99, p.273-279, 2005.
• BADR, A.; EL-SHAZLY, H.H.; HELAIL, N.S.; EL GHANIM, W. Genetic diversity of Artemisia populations in central and north Saudi Arabia based on morphological variation and RAPD polymorphism. Plant Syst. Evol., v.298, p.871-886, 2012.
• BESHAY, E. Therapeutic efficacy of Artemisia absinthium against Hymenolepis nana: in vitro and in vivo studies in comparison with the anthelmintic praziquantel. J. Helminthol., v.92, p.298-308, 2018.
• BOONMASAWAI, S.; SUNGPRADIT, S.; JIRAPATTHARASATE, C. et al. Effects of alcoholic extract from pomegranate (Punica granatum L.) peels on gastrointestinal nematode egg counts in doe. J. Appl. Anim. Sci., v.6, n.2, 2013.
• CÁCERES, A.L.; FLORES-GIUBI, M.E.; ROMERO-RODRÍGUEZ, M.C.; ALVARENGA, N.L. In vitro anthelmintic activity and chemical composition of methanol extracts and fractions of Croton paraguayensis and Vernonia brasiliana against Eisenia fetida. Asian Pac. J. Trop. Dis., v.7, p.71-74, 2017.
• CRESPO-ORTIZ, M.P.; WEI, M.Q. Antitumor activity of artemisinin and its derivatives:. from a well-known antimalarial agent to a potential anticancer drug. Biomed Res. Int., v.2012,| p.247597, 2012.
• CURICO, G.; GARCÍA-BARDALES, P.; PINEDO, T. et al. Resistance to single dose albendazole and reinfection with intestinal helminths among children ages 2 to 11 years from the Peruvian Amazon region: a study protocol. BMC Infect. Dis., v.22, p.528, 2022.
• DIAZ, E.; MONDRAGON, J.; RAMIREZ, E.; BERNAL, R. Epidemiology and control of intestinal parasites with nitazoxanine in children in Mexico. Am. J. Trop. Med. Hyg., v.68, p.384-385, 2003.
• DRURY, R.; WALLINGTON, E. Tissue histology. In: CARLETON, H.M.; DRURY, R.; WALLINGTON, E. Carleton’s histological techniques. New York: Oxford University Press, 1973. 520p.
• EFFERTH, T. Molecular pharmacology and pharmacogenomics of artemisinin and its derivatives in cancer cells. Curr. Drug Targets, v.7, p.407-421, 2006.
• EL-MAGGAR, E. Artemisia herba alba & Artemisia monosperma: the discovery of the first potential Egyptian plant sources for the pharmaceutical commercial production of artemisinin and some of its related analogues. J. Appl. Pharm. Sci., v.2, p.77-91, 2012.
• EMERY, D.L.; HUNT, P.W.; LE JAMBRE, L.F. Haemonchus contortus: the then and now, and where to from here? Int. J. Parasitol., v.46, p.755-769, 2016.
• FISSIHA, W.; KINDE, M.Z. Anthelmintic resistance and its mechanism: a review. Infect. Drug. Resist., v.14, p.5403-5410, 2021.
• GITHIORI, J.B.; HÖGLUND, J.; WALLER, P.J.; BAKER, R.L. The anthelmintic efficacy of the plant, Albizia anthelmintica, against the nematode parasites Haemonchus contortus of sheep and Heligmosomoides polygyrus of mice. Vet. Parasitol., v.116, p.23-34, 2003.
• GOODMAN, A.G. Ethyl alcohol. In: BRUNTON, L.L.; HILAL DANDAN, R.; KNOLLMANN, B.C. The Phamacological basis of therapeutics. United States: Macmillan and Co., 1960. [357p.].
• HO, W.E.; PEH, H.Y.; CHAN, T.K.; WONG, W.F. Artemisinins: pharmacological actions beyond anti-malarial. Pharmacol. Ther., v.142, p.126-139, 2014.
• HOU, J.; WANG, D.; ZHANG, R.; WANG, H. Experimental therapy of hepatoma with artemisinin and its derivatives: in vitro and in vivo activity, chemosensitization, and mechanisms of action. Clin. Cancer Res., v.14, p.5519-5530, 2008.
• HUSEN, E.A.; TAFESSE, G.; HAJARE, S.T. et al. Cross-sectional study on assessment of frequency of intestinal helminth infections and its related risk factors among school children from Adola Town, Ethiopia. BioMed. Res. Int., v.2022, p.5908938, 2022.
• IRSHAID, F.; MANSI, K.; ABURJAI, T. Antidiabetic effect of essential oil from Artemisia sieberi growing in Jordan in normal and alloxan induced diabetic rats. Pak. J. Biol. Sci., v.13, p.423-430, 2010.
• JESÚS-MARTÍNEZ, D.; OLMEDO-JUÁREZ, A.; OLIVARES-PÉREZ, J. et al. In vitro anthelmintic activity of methanolic extract from Caesalpinia coriaria J. Willd fruits against Haemonchus contortus eggs and infective larvae. Biomed. Res. Int., v.2018, p.7375693, 2018.
• KABOUTARI, J.; HAYDARNEJAD, M.S.; FATAHIAN, D.R.A.; RAEISI, V.S. Histomorphometric study on the effects of Artemisia sieberi extract in mice skin. J. HerbMed. Pharmacol., v.4, p.20-24, 2015.
• KANTHAL, L.K.; SATYAVATHI, K.; NAIDU, A. et al. Pharmacological potential of lactuca runcinata dc: a review. Int. J. Pharm. Pharm. Sci., v.4, n.4, 2014.
• KERBOEUF, D.; RIOU, M.; GUÉGNARD, F. Flavonoids and related compounds in parasitic disease control. Mini Rev. Med. Chem., v.8, p.116-128, 2008.
• KHALAJI, S.; ZAGHARI, M.; HATAMI, K. et al. Black cumin seeds, Artemisia leaves (Artemisia sieberi), and Camellia L. plant extract as phytogenic products in broiler diets and their effects on performance, blood constituents, immunity, and cecal microbial population. Poult. Sci., v.90, p.2500-2510, 2011.
• KREPEL, H.; HARING, T.; BAETA, S.; POLDERMAN, A. Treatment of mixed Oesophagostomum and hookworm infection: effect of albendazole, pyrantel pamoate, levamisole and thiabendazole. Trans. R. Soc. Trop. Med. Hyg., v.87, p.87-89, 1993.
• LIRA, C.D.; BARRY, T.; POMROY, W. et al. Willow (Salix spp.) fodder blocks for growth and sustainable management of internal parasites in grazing lambs. Anim. Feed Sci. Technol., v.141, p.61-81, 2008.
• LOO, C.S.N.; LAM, N.S.K.; YU, D. et al. Artemisinin and its derivatives in treating protozoan infections beyond malaria. Pharmacol. Res., vv.117, p.192-217, 2017.
• LÜ, G.; ZHANG, W.; WANG, J. et al. Application of a cDNA microarray for profiling the gene expression of Echinococcus granulosus protoscoleces treated with albendazole and artemisinin. Mol. Biochem. Parasitol., v.198, p.59-65, 2014.
• MANIKANDAN, P.; LETCHOUMY, P. V.; GOPALAKRISHNAN, M.; NAGINI, S. J. F. Evaluation of Azadirachta indica leaf fractions for in vitro antioxidant potential and in vivo modulation of biomarkers of chemoprevention in the hamster buccal pouch carcinogenesis model. Food Chem. Toxicol., v.46, p.2332-2343, 2008.
• MILLER, L.H.; SU, X. Artemisinin: discovery from the Chinese herbal garden. Cell, v.146, p.855-858, 2011.
• MORGAN, E.; ARNOLD, M.; GINI, A. et al. Global burden of colorectal cancer in 2020 and 2040: incidence and mortality estimates from GLOBOCAN. Gut, v.72, p.338-344, 2023.
• NAHREVANIAN, H.; SHEYKHKANLOOYE MILAN, B.; KAZEMI, M. et al. Antimalarial effects of Iranian flora Artemisia sieberi on Plasmodium berghei in vivo in mice and phytochemistry analysis of its herbal extracts. Malar. Res. Treat., v.2012, p.727032, 2012.
• NASR, F.A.; NOMAN, O.M.; MOTHANA, R.A. et al. Cytotoxic, antimicrobial and antioxidant activities and phytochemical analysis of Artemisia judaica and Artemisia sieberi in Saudi Arabia. Afr. J. Pharm. Pharmacol., v.14, p.278-284, 2020.
• NEGAHBAN, M.; MOHARRAMIPOUR, S.; SEFIDKON, F. Insecticidal activity and chemical composition of Artemisia sieben besser essential oil from Karaj, Iran. J. Asia. Pac. Entomol., v.9, p.61-66, 2006.
• PADOSCH, S.A.; LACHENMEIER, D.W.; KRÖNER, L.U. Absinthism: a fictitious 19th century syndrome with present impact. Subst .Abuse Treat. Prev. Polic., v.10, p.1-14, 2006.
• PARIDA, S.; PATRO, V.J.; MISHRA, U.S. et al. Anthelmintic potential of crude extracts and its various fractions of different parts of Pterospermum acerifolium Linn. Int. J. Pharma. Sci. Rev. Res., v.1, p.107-111, 2010.
• SAGAR, S.; YANCE, D.; WONG, R. Natural health products that inhibit angiogenesis: a potential source for investigational new agents to treat cancer-part 1. Curr. Oncol., v.13, p.14-26, 2006.
• SALEH, N.; REWAIDA, A.; AFAF, A. et al. Anthelmintic activity of Artemisia monosperma methanol extracts against Eisenia fetida in vitro study. Indian J. Anim. Res., v.58, p.115-121, 2024.
• SALIH, A.M.; QAHTAN, A.A.; AL-QURAINY, F. Phytochemicals identification and bioactive compounds estimation of Artemisia Species grown in Saudia Arabia. Metabolites, v.13, p.443, 2023.
• SILVA, N.; GUYATT, H.; BUNDY, D. Anthelmintics: a comparative review of their clinical pharmacology. Drugs, v.53, p.769-788,1997.
• ŠKROVÁNKOVÁ, S.; MIŠURCOVÁ, L.; MACHŮ, L. Antioxidant activity and protecting health effects of common medicinal plants. Adv. Food Nutr. Res., v.67, p.75-139, 2012.
• STEPEK, G.; BEHNKE, J.M.; BUTTLE, D.J.; DUCE, I.R. Natural plant cysteine proteinases as anthelmintics? Trends Parasitol., v.20, p.322-327, 2004.
• SUNG, H.; FERLAY, J.; SIEGEL, R.L. et al. Global cancer statistics. 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin., v.71, p.209-249, 2021.
• TANSATIT, T.; SAHAPHONG, S.; RIENGROJPITAK, S. et al. Fasciola gigantica: the in vitro effects of artesunate as compared to triclabendazole on the 3-weeks-old juvenile. Exp. Parasitol., v.131, p.8-19, 2012.
• THE WORLD health report 2005: make every mother and child count. World Health Organization, 2005.
• TORRES-ACOSTA, J.; MOLENTO, M.; DE GIVES, P.M. Research and implementation of novel approaches for the control of nematode parasites in Latin America and the Caribbean: is there sufficient incentive for a greater extension effort?. Vet. Parasitol., v.186, p.132-142, 2012.
• YANG, L.; WEN, K.S.; RUAN, X. et al. Response of plant secondary metabolites to environmental factors. Molecules, v.23, p.762, 2018.

Publication Dates

• Publication in this collection
  21 Feb 2025
• Date of issue
  Mar-Apr 2025

History

• Received
  09 May 2024
• Accepted
  19 July 2024