Artemisia afra Toxicology

Artemisia afra Jacq.

Artemisia afra Toxicology

Artemisia afra Jacq.

2005

James Tshikosa Mukinda
Acute and chronic toxicity of the flavonoid- containing plant, Artemisia afra in rodents
A thesis submitted in partial fulfillment of the requirements for the degree of Magister Scientiae in the Department of Pharmacology, University of the Western Cape. December 2005

Acute and chronic toxicity of the flavonoid- containing plant, Artemisia afra in rodents

Abstract :

Despite being one of the most popular traditional plant medicines used in South Africa, very little is known about the safety and efficacy of Artemisia afra. The aim of this study was to investigate the possible toxicity of the flavonoid-containing plant, Artemisia afra and especially to establish the safety of the aqueous extract of this plant after acute and chronic administration to mice and rats respectively. To realize this objective, the following were determined : (1) the adverse effects caused by the dried aqueous extract of Artemisia afra in mice and rats, (2) the LD 50 for the aqueous extract of the plant administered acutely in mice using the traditional method of Litchfield and Wilcoxon and a computer-based (AOT425statPgm) method, (3) the doses (i.e. LD 50 ) of the plant causing adverse effects after oral and intraperitoneal acute administration, (4) the general toxicity profile obtained after chronic oral dosing with the extract in rats and (5) the correlation between the plasma levels of luteolin, plant doses and toxic effects obtained after the chronic oral administration of the plant extract to rats. It was hypothesized that : (i) the flavonoids such as luteolin contained in Artemisia afra, if given in high enough doses may be associated with some adverse effects ; (ii) the dose of the plant at which such adverse effects occurred would depend on both the route of administration (oral vs. intraperitoneal) as well as the duration (acute or chronic) of exposure to the plant material ; (iii) the new computer program-based method (i.e. AOT425statPgm) for testing acute toxicity of environmental pollutants, can advantageously be used to evaluate the acute toxicity of A. afra extract, and (iv) plasma levels of luteolin could be used as a marker to monitor A. afra toxicity and/or its bioavailability (ingestion) in rats.

Plant material (i.e. leaves & aerial parts) was collected from Montague Museum, Western Cape Province, South Africa and the freeze-dried aqueous extract prepared. Mice were administered single oral doses of A. afra (0.175 to 24 g/kg) or intra-peritoneal doses (0.175 to 5,5g/kg) and were monitored for mortality and toxic symptoms for two weeks. The LD 50 values for each route were determined according to the methods of “Litchfield and Wilcoxon” and the “Acute Oral Toxicity (Guideline 425) statistical program” (AOT425statPgm). In another study, rats were given oral doses (0,1 or 1g/kg) of A. afra for : (a) three months (92 days), after which blood was withdrawn for measurement of several haematological and biochemical parameters and plasma levels of luteolin by HPLC assay, and selected tissues inspected for histopathological changes, or (b) seven days, after which the levels of luteolin in the blood of rats were measured. In addition, the luteolin level in the Artemisia afra aqueous extract was determined using HPLC.

The LD 50 of A. afra extract after acute intraperitoneal injection in mice was 27% of that after oral administration (i.e. 2450 mg/kg vs. 8960mg/kg). The AOT425statPgm method, while giving the same symptom profile and LD 50 results and having the same study time to conduct, however involved substantially fewer mice, in fact only 20 and 50% of that used in the Litchfield and Wilcoxon method for the acute p.o. and i.p. toxicity testing, respectively. In the chronic toxicity study, all the rats survived the duration of the treatment (i.e. LD 50 was much higher than 1000mg/kg), with no significant changes in physical signs, haematological and biochemical parameters, except for a transient decrease in aspartate aminotransferase (AST) activity. There were also no significant differences in the organs weights, and the histopathological results showed normal architecture suggesting no morphological disturbances. The luteolin levels in the Artemisia afra and rat plasma were 0,923± 0,015 µg/mg extract and less than 0,2 µg/ml plasma, respectively.

Collectively, the results indicate that acute doses of A. afra are relatively non-toxic in mice irrespective of the route of administration used, chronic doses of A. afra are very safe in the rat, that the AOT425statPgm is a potentially useful tool for the evaluation of acute toxicity of plant medicines and, finally, that luteolin may be a good marker for A. afra bioavailability, but not likely for its toxicity.

Keywords : Acute and chronic toxicity, Artemisia afra, Hydrolyzed extract, Flavonoids, Luteolin, HPLC, Rodents, Adverse effects, Oral and intra-peritoneal routes, Heamatological parameters, Biochemical parameters, Histopathological examination

2007

J.T. Mukinda, J.A. Syce
Acute and chronic toxicity of the aqueous extract of Artemisia afra in rodents
Journal of Ethnopharmacology 112 (2007) 138–144

Full text submitted to request

Abstract

Artemisia afra (Jacq. Ex. Willd), “African Wormwood” is widely used traditionally in South Africa with no literature evidence substantiating its safety. The aim of this study was to investigate the safety of the aqueous extract of Artemisia afra by determining its pharmaco-toxicological effects after acute and chronic administration in mice and rats, respectively. The aqueous extract mimicked the traditional decoction dosage form of Artemisia afra. In mice, single intraperitoneal injections of Artemisia afra-extract (1.5–5.5g/kg) induced a regular dose-dependent increase in the death rate and incidence of general behaviour adverse effects, while with single oral doses (2–24g/kg) the increases in incidence of general behaviour adverse effects and mortality rate were dose-independent. The LD 50s after acute intraperitoneal and oral doses were 2.45 and 8.96g/kg, respectively.Rats given oral doses of Artemisia afra-extract (0.1or1g/kg/day) survived the 3 months of dosing (i.e.LD 50 much highert han1g/kg), experienced no significant changes in general behaviour and haematological and biochemical parameters, except for transient decrease in AST activity. No significant changes were observed in organ weights, and histopathological results showed normal profile suggesting no morphological alterations. Collectively, the results indicate that Artemisia afra-extract is non-toxic when given acutely, has low chronic toxicity potential and, in high doses, may have a hepatoprotective effect.

Keywords : Artemisia afra ; Aqueous extract ; Plant medicine ; Acute and chronic toxicity ; Rodents ; Traditional dosage form

***

James Waweru Gathirwa, Geoffrey M. Rukunga, Eliud N. M. Njagi, Sabah A. Omar, Anastasia N. Guantai, Charles N. Muthaura, Peter G. Mwitari, Cecilia W. Kimani, Peter G. Kirira, Festus M. Tolo, Teresia N. Ndunda, Isaiah O. Ndiege
In vitro anti-plasmodial and in vivo anti-malarial activity of some plants traditionally used for the treatment of malaria by the Meru community in Kenya
Journal of Natural medicine (2007) 61:261–268

Abstract :

Extracts of seven medicinal plant species used for treatment of malaria in traditional/cultural health systems of the Ameru people in Kenya were tested in vitro and in vivo against Plasmodium falciparum (D6 and W2 strains) and P. berghei, respectively. Of the plants tested, 28.57% were highly active (IC 50 <10 lg/ml) and 42.86% moderately active (IC 50 10–50 lg/ml), while 28.57% had weak activity of 50–125 lg/ml in vitro. The water and methanol extracts of Boscia salicifolia Oliv. and Artemisia afra Jacq. (ex-Willd.) were the most active against both the chloroquine (CQ)-sensitive (D6) and the CQ-resistant (W2) P. falciparum strains. Artemisia afra and Rhus natalensis Bernh. (ex-Krauss) exhibited the highest parasite clearance and chemo-suppression (>70%) in vivo (in mice). The plants with high in vitro anti-plasmodial (low IC 50 values) and high anti-malarial activity (high chemo-suppression) in vivo are potential sources of novel anti-malarial drugs.

Keywords : Anti-malarial, Anti-plasmodial, Toxicity, Boscia salicifolia, Artemisia afra, Rhus natalensis

2012

Andrea Lubbe, Isabell Seibert, Thomas Klimkait, Frank van der Kooy
Ethnopharmacology in overdrive : The remarkable anti-HIV activity of Artemisia annua (and Artemisia afra)
Journal of Ethnopharmacology (2012)

Full text submitted to request

Abstract :

Ethnopharmacological relevance : Artemisia annua contains the well-known antimalarial compound artemisinin, which forms the backbone of the global malaria treatment regime. In African countries a tea infusion prepared from Artemisia annua has been used for the treatment of malaria only for the past 10–20 years. Several informal claims in Africa exist that the Artemisia annua tea infusions are also able to inhibit HIV. Since HIV is a relatively newly emerged disease, the claims, if substantiated, could provide a very good example of “ethnopharmacology in overdrive”. The objective of this study was to provide quantitative scientific evidence that the Artemisia annua tea infusion exhibits anti-HIV activity through in vitro studies. A second objective was to determine if artemisinin plays a direct or indirect (synergistic) role in any observed activity. This was done by the inclusion of a chemically closely related species, Artemisia afra, known not to contain any artemisinin in our studies.

Materials and Methods : Validated cellular systems were used to test Artemisia annua tea samples for anti-HIV activity. Two independent tests with different formats (an infection format and a co-cultivation format) were used. Samples were also tested for cellular toxicity against the human cells used in the assays.

Results : The Artemisia annua tea infusion was found to be highly active with IC 50 values as low as 2.0 ?g/mL. Moreover we found that artemisinin was inactive at 25 ?g/mL and that a chemically related species Artemisia afra (not containing artemisinin) showed a similar level of activity. This indicates that the role of artemisinin, directly or indirectly (synergism), in the observed activity is rather limited. Additionally, no cellular toxicity was seen for the tea infusion at the highest concentrations tested.

Conclusion : This study provides the first in vitro evidence of anti-HIV activity of the Artemisia annua tea infusion. We also report for the first time on the anti-HIV activity of Artemisia afra although this was not an objective of this study. These results open the way to identify new active pharmaceutical ingredients in Artemisia annua and thereby potentially reduce the cost for the production of the important antimalarial compound artemisinin.

Keywords : Artemisia afra, Artemisia annua, Artemisinin, HIV, Malaria Tea infusion

2013

L. Spies, T.C. Koekemoer, A.A. Sowemimo, E.D. Goosen, M. Van de Venter
Caspase-dependent apoptosis is induced by Artemisia afra Jacq. ex Willd in a mitochondria-dependent manner after G2/M arrest
South African Journal of Botany, Volume 84, 2013, pp. 104-109

Caspase-dependent apoptosis is induced by Artemisia afra Jacq. ex Willd in a mitochondria-dependent manner after G2/M arrest

Abstract

Artemisia afra is one of the oldest, most well known and widely used traditional medicinal plants in South Africa. It is used to treat many different medical conditions, particularly respiratory and inflammatory ailments (Liu et al., 2009). There is no reported evidence of its use for the treatment of cancer but due to its reported cytotoxicity (Fouche et al., 2008 ; Mativandlela et al., 2008), we investigated the mode of cell death induced by an ethanolic A. afra extract by using two cancer cell lines. IC50 values of 18.21 and 31.88 μg/mL of ethanol extracts were determined against U937 and HeLa cancer cells, respectively. An IC50 value of the aqueous extract was greater than 250 μg/mL. The effect of the cytotoxic ethanolic A. afra extract on U937 and HeLa cells and their progression through the cell cycle, apoptosis and mitochondrial membrane potential were investigated. Melphalan was used as a positive control. After 12 h of treatment with A. afra a delay in G2/M phase of the cell cycle was evident. Apoptosis was confirmed by using the TUNEL assay for DNA fragmentation, as well as fluorescent staining with annexin V-FITC. Apoptosis was evident with the positive control and A. afra treatment at 24 and 48 h. JC-1 staining showed a decrease in mitochondrial membrane potential at 24 h. The results obtained suggest that A. afra potentially has medicinal anticancer properties

Keywords : Artemisia afra, Cytotoxicity, Apoptosis, HeLa, U937

2015

Nikodimos Eshetu
Evaluation of the acute and sub-chronic toxic effects of aqueous leaves extracts of Artemisia afra on Liver, Kidney and some Blood parameters in Wistar Rats
A thesis submitted to the Department of Anatomy, School of medicine, College of health science, Addis Ababa University, Addis Ababa, Ethiopia in partial fulfillment of the requirement for the Degree of Master of science (MSc) in anatomy. Addis Ababa, Ethiopia August 2015.

Evaluation of the Acute and Sub-chronic Toxic Effects of Aqueous Leaf Extracts of {Artemisia afra} on Liver, Kidney and Some Blood Parameters in Wistar Rats

Abstract

Traditional medicine has remained to be the most affordable and easily accessible source of treatment in the primary healthcare system of resource poor communities and, it is the only means of treatment for such communities. Plants have been the basis for treatment throughout human history, and they are still widely practiced today as part of traditional medicine. The plant Artemisia afra has been shown to display a wide spectrum of biological and pharmacological activities, which provide experimental support for the empiric ethno-pharmacological use of this plant in traditional medicine. However, despite its widespread use, very little is known about its safety and efficacy.

This Experimental laboratory based study has been carried out to investigate the acute and subchronic toxic effects of leaves of Artemisia afra on liver and kidney ; and some blood parameters in rats. The study was carried out in Department of Anatomy, School of Medicine, Addis Ababa University. The experiments were performed on 59 Wister rats (32 for acute and 27 for subchronic study) based on the OECD guideline they were assigned to each group randomly. The study was conducted from January 2014-July 2015. Various doses of aqueous extract of the leaves were employed for single dose toxicity studies, while the effective dose and triple the effective dose were used for repeated toxicity studies.

This study showed that the oral lethal dose (LD 50s ) is higher than 5000mg/kg. Generally in the acute toxicity study ; the general behavior and body weights were not altered in Wister rats administered with doses up to 5000mg/kg. After subchronic study with both doses (600 and 1800mg/kg), there were no significant changes in the overall body weight, the evaluated hematological and most of the biochemical parameters. No death was recorded. In gross observation, the kidneys and liver of treatment groups appear normal in their texture, size or color as compared to the control. Histopathological presentations were generally normal
though there were mild mononuclear leukocytic infiltrations around the central venules & portal areas of Wister rats’ liver at both 600 and 1800mg/kg dose in addition minor tubulointerstitial leukocytic infiltrations were observed in small areas of kidney sections administered higher dose. Findings of this study revealed that A. afra is relatively safe.

Keywords : Artemisia afra, Traditional medicine, Toxicological assessment

***

Mekonen, Ketema
Evaluation of the Acute and Subacute Toxicity of Aqueous Leaves Extracts of Artemisia Afra on Brain, Heart and Suprarenal Gland in Swiss Albino Mice
Thesis submitted to the Department of Anatomy, School of medicine, College of health science, Addis Ababa University, Addis Ababa, Ethiopia in partial fulfillment of the requirement for the Degree of Master of science (Msc) in anatomy. August, 2015

Evaluation of the Acute and Subacute Toxicity of Aqueous Leaves Extracts of Artemisia Afra on Brain, Heart and Suprarenal Gland in Swiss Albino Mice

Résumé :

Having primary health care is a human right which is fulfilled by western country because of expansion of health infrastructure, and increased quantity and quality of health professionals. But many developing countries including Ethiopia are yet far from achieving this. In Ethiopia, the majority of population relay on traditional medicine as a source of health care. The most common sources of traditional medicine are plants. A.afra is one of these plants that are used to treat different aliments. The aim of the present study was to investigate the toxic effects of A.afra on brain, heart and suprarenal glands. The study was conducted at Addis Ababa University, College of Health Science, School of Medicine, Department of Anatomy & Department of Physiology from Jan, 2014 to July, 2015. The plant was collected from Bale National park in Oromia Regional State. The plant was air dried and aqueous extract was prepared. In this research a total of 54 male and female mice of 8-12 weeks of age weighing 25-30g were used. The extract was given by oral rout in both acute and subacute study. The doses for acute toxicity study were 200mg/kg, 700mg/kg, 1200mg/kg, 2200mg/kg, 3200mg/kg, 4200mg/kg and 5000mg/kg of body weight, while for subacute toxcicy study a doses of 600mg/kg(low dose) and 1800mg/kg(high dose) of body weight were used. LD50 was grounded to be greater or above 5000mg/kg which indicates that the plant is relatively safe. There were no observed signs of toxicity at the lower three doses, although mild toxicity sign was observed at the higher dose level in dose dependent manner. In the subacute study, two treatment groups 600mg/kg and 1800mg/kg and one control group containing both sexes were used. Weights of mice were measured weekly and individual mice were observed for possible toxicity sign. At the end of 28 days, the animals were scarified and organs were harvested and processed for microscopic examination. No toxicity signs were observed in all treatment groups. There were also no significant weight changes between the treated and control group. On microscopic examination of the brain, heart and suprarenal glands no sign of cellular injury was observed. From this study it can be concluded that A.afra is relatively safe in mice.

Key words : Traditional medicine, A.afra, Toxicity study, LD50, Histopathology

2016

Nikodimos Eshetu Mekbeb Afework, Eyasu Makonnen, Asfaw Debella, Wondwossen Ergete, Tesfaye Tolesss
Evaluation of the Acute and Sub-chronic Toxic Effects of Aqueous Leaf Extracts of Artemisia afra on Liver, Kidney and Some Blood Parameters in Wistar Rats
Advances in Bioscience and Bioengineering, Vol. 1, No. 1, 2016, pp. 1-9

Abstract :

Background : Artemisia afra is a plant traditionally used for treatment of different diseases in many parts of the world including Ethiopia. Its effects on different organs, however, have not yet been investigated. The objective of the present study was, therefore, to evaluate the acute and sub-chronic toxic effects of aqueous leaf extracts of Artemisia afra on Liver, Kidney and some Blood parameters in Rats.

Methods : For acute toxicity study, aqueous extracts of the leaves were administered in a single dose of 200, 700, 1200, 2200, 3200, 4200 and 5000mg/kg body weight, while the low dose (600mg/kg) and triple of lower dose (1800mg/kg) were used for sub-chronic toxicity studies. Selected hematological and biochemical parameters of the blood followed by histopathological analysis were investigated after 90 days of daily administrations. The results were expressed as M ± SE, and differences at P < 0.05 were considered significant. Differences between the experimental and control groups were analyzed using one-way analysis of variance (ANOVA), followed by Dunnett’s T-test to determine their level of significance.

Results : The current study showed that the median oral lethal dose (LD50) was greater than 5000mg/kg. Acute toxicity study revealed some changes in general behavior of the rats above 3200mg/kg. The levels of blood parameters did not change though AST level decreased significantly in female animals after 90 days of sub-chronic treatment with 1800mg/kg. Histopathological presentations were generally normal though there were mild mononuclear leukocytic infiltrations around the central venules & portal areas of rats’ liver at both 600 and 1800mg/kg dose. Furthermore, minor tubulointerstitial leukocytic infiltrations were observed in small areas of kidney sections treated at higher dose.

Conclusion : The aqueous extract of Artemisia afra at the test doses did not show significant toxicity : the minor inflammatory changes observed in this study were not accompanied by significant change in any of the hematological and biochemical markers of liver injury. It might be a response to parenchymal cell death with causes ranging from infectious agents, exposure to toxicants, generation of toxic metabolites, and tissue anoxia.

Keywords : Artemisia afra, Traditional Medicine, Toxicological Assessment

2019

Ndeye Fatou Kane, Mutinda Cleophas Kyama, Joseph Kangethe Nganga, Ahmed Hassanali, Mouhamadou Diallo, Francis Thuo Kimani
Acute Toxicity Effect of Artemisia afra Plant Extracts on the Liver, Kidney, Spleen and in Vivo Antimalarial Assay on Swiss Albino Mice
Advances in Bioscience and Bioengineering. Vol. 7, No. 4, 2019, pp. 64-71.

Full text submitted to request

Abstract :

Artemisia afra (Jacq. Ex. Wild), or "African Wormwood" belonging to the family of Astereaces and is widely used traditionally for health care in the eastern part of Africa with few research evidence substantiating its safety. The aim of this study was to investigate the safety of the ethanolic, dichloromethane, and hexanolic extracts of Artemisia afra by determining its pharmaco-toxicological effects after an acute oral administration in mice and to test also their in vivo antimalarial effects. Oral acute doses of Artemisia afra extracts were given to thirty mice at the doses of 1000, 2000 and 2500 mg/kg of body weight. The mice were then observed for fourteen days, toxicity signs, body weight, organs weight and biochemical parameters were checked. Four days peter’s test was run on mice to determine the in vivo antimalarial activity of the plant extracts and the IC50 for each extract was determined. The results show few toxicity signs from the first two days after oral administration. There were no differences in organs weight and body weight for the experimental mice when compared to the control group. The level of alanine transaminase (ALT) and aspartate transaminase (AST) were found do not be statistically different from the control. The LD 50 of the extracts was found to be greater than 2500 mg/kg of body weight. The results also showed a high antimalarial effect of the extracts when tested in vivo using Plasmodium Berghei Anka. In Conclusion Artemisia afra is a strong drug candidate for malaria with no toxic effects in high dosage.

Keywords : Oral Acute Toxicity, Medicinal Plant, Antimalarial Assay, Plasmodium Berghei Anka, Artemisia afra, Biochemical Test

***

Ndeye Fatou Kane
Effect of extracts of Artemisia afra collected from five different regions in Africa (Kenya, Burundi, Tanzania, South Africa and Senegal) on in vitro and in vivo cultures of Plasmodium Species
A thesis submitted to Pan African University Institute of Science, Technology and Innovation in partial fulfillment of the requirement for the award of the degree of Doctor of Philosophy in Molecular Biology and Biotechnology of the Pan African University, 2019

Full text submitted to request

Abstract :

Malaria is one of the deadliest disease in the world affecting millions of individuals yearly. Artemisinin combination therapies (ACTs), which are the first line of defense against this disease for many years, show some inefficacy due to delay in parasite clearance. Many episodes of resistance against these drugs have been registered in many countries in Africa and in Asia. Currently, there is growing research interest in the use of full blend extract of medicinal plants like Artemisia afra or Artemisia annua as an alternative treatment. Artemisia afra is an indigenous species to Africa, and are traditionally used for decades by traditional healers to cure a lot of afflictions among them malaria. The main objective of this thesis is to compare the growth inhibition effects and molecular profiles of the parasite exposed to extracts of the Artemisia afra plants collected from five regions. Artemisia afra leaves were collected from five countries in Africa (South Africa, Tanzania, Burundi, Senegal, and Kenya), and compared for their level of phytochemicals content, antimalarial, and antioxidant activities. The plant extracts were first tested in vitro and the most active extracts were incubated during 2 days with the parasites to study the expression level of FabI and FabZ. A virtual screening was ran using PyRx with vina to determine the potential interactions between the Fab enzymes (FabI and FabZ) and the active compounds of the plant extracts. The result showed a big antimalaria

l property of the plant, however a different level of activity depending on the geographical localization. A. afra collected from Burundi was found to have the highest level of phenols and flavonoids. This plant also exhibited the highest antioxidant and antimalarial activities compared to the others. Acute toxicity test run in mice revealed an ED50 greater than 2500mg/kg body weight and no toxic sign was detected on the liver, the organs, and the tissues. The Fab I enzyme, which plays important rule during the liver stage of the malaria infection was found to be downregulated in the W2 strain, after exposition of the parasites with the ethanolic and dichloromethane of the plant extract collected from Burundi. In the D6 strain the enzyme was downregulated with the hexane and ethanolic extracts of the plant. Compared to Fab I, Fab Z was found to be downregulated only on the D6 strain when exposed to the hexane and ethanolic extracts, both extracts downregulate Fab I and Fab Z in the two strains (D6, W2). Virtual screening showed interaction between the active compounds of the plant and the Fab enzymes. In conclusion the results confirm the high antimalarial effect of Artemisia afra and also its prophylactic effect with the inhibition of Fab I enzyme which is crucial during liver stage of the plasmodium falciparum life cycle. The difference in level of flavonoids and phenols suggest that the agro-ecological zone play an important role in influencing the level of phytochemical. Artemisia afra active compounds detected during GCMS analysis was found to interact with the fab enzymes of the parasite, further study need to be done to confirm their in vitro activities of those active compounds.

Published online by La vie re-belle
 6/04/2020
 http://lavierebelle.org/artemisia-afra-toxicologie

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