Dear Label_Name,

Greetings! I hope you are well.

I am writing on behalf of an international publishing house, Science and Education Publishing, USA (http://www.sciepub.com).

Science and Education Publishing (SciEP) is a multidisciplinary open access publisher of Journals covering the fields of Science, Technology and Medicine. All SciEP publications are subject to high-quality peer review, editorial and production processes and are freely available online.

Welcome to the April, 2013 edition of SciEP newsletter, providing monthly summaries of articles on science & education related topics of interest to scientist and professionals. The mission of our journals is to provide an open access forum for publishing new findings on issues pertinent to scientific and academic field. We also invite you to submit original research papers, review articles, short reports, or case studies are considered for publication in our Multidisciplinary Journals.

American Journal of Public Health Research
DOI: 10.12691/ajphr-1-1-3

The Implementation of a Tele-Homecare System with Service Oriented Architecture and HL7 Message Transmission Standard
Ching-Sung Wang

American Journal of Medical and Biological Research
DOI: 10.12691/ajmbr-1-1-2

The Influence of Ethanol on Pyruvate Kinases Activity in Vivo, in Vitro, in Silico
Lelevich Sergey Vladimirovich, Khrustalev Vladislav Victorovich, Barkovsky Eugene Victorovich, Shedogubova Tatyana Aleksandrovna

American Journal of Medical Sciences and Medicine
DOI: 10.12691/ajmsm-1-1-1

Clinical and Histological Features in Wegener Granulomatosis
Li Cavoli Gioacchino, Passantino Rita, Bono Luisa, Tortorici Calogera, Ferrantelli Angelo, Giammarresi Carlo, Rotolo Ugo

American Journal of Infectious Diseases and Microbiology
DOI: 10.12691/ajidm-1-1-2

The Curse of Lymphatic Filariasis: Would the Continual Use of Diethylcarbamazine Eliminate this Scourge in Papua New Guinea?
Anne Lanham, Lillian Mwanri

We have released issues of some journals on February 2013.
American Journal of Public Health Research
American Journal of Medical Sciences and Medicine
American Journal of Medical and Biological Research
American Journal of Pharmacological Sciences
American Journal of Clinical Medicine Research
American Journal of Nursing Research
American Journal of Cancer Prevention
American Journal of Medicine Studies
American Journal of Infectious Diseases and Microbiology
American Journal of Cardiovascular Disease Research
American Journal of Biomedical Research
American Journal of Mechanical Engineering
American Journal of Microbiological Research
Journal of Preventive Medicine
American Journal of Food and Nutrition
American Journal of Hypertension Research
American Journal of Sports Science and Medicine
Journal of Cancer Research and Treatment
American Journal of Zoological Research
Journal of Biomedical Engineering and Technology
American Journal of Microbiological Research
Biomedical Science and Engineering

We are pleased to announce that submissions are now being accepted for peer review for all the journals across all our subject areas in SciEP.

Please submit your manuscript via our online submission system:
http://www.sciepub.com/author/signin

To unsubscribe from our mailing list, please click on the following link: unsubscribe

I am looking forward to hearing from you.

Kind regards,

David Rain
Email:join@sciepub.com
Science and Education Publishing (http://www.sciepub.com)

Label_Title

Dear Label_Name,

Greetings! I hope you are well.

I am writing on behalf of an international publishing house, Science and Education Publishing, USA (http://www.sciepub.com).

Science and Education Publishing (SciEP) is a multidisciplinary open access publisher of Journals covering the fields of Science, Technology and Medicine. All SciEP publications are subject to high-quality peer review, editorial and production processes and are freely available online.

Welcome to the April, 2013 edition of SciEP newsletter, providing monthly summaries of articles on science & education related topics of interest to scientist and professionals. The mission of our journals is to provide an open access forum for publishing new findings on issues pertinent to scientific and academic field. We also invite you to submit original research papers, review articles, short reports, or case studies are considered for publication in our Multidisciplinary Journals.

American Journal of Public Health Research
DOI: 10.12691/ajphr-1-1-3

The Implementation of a Tele-Homecare System with Service Oriented Architecture and HL7 Message Transmission Standard
Ching-Sung Wang

American Journal of Medical and Biological Research
DOI: 10.12691/ajmbr-1-1-2

The Influence of Ethanol on Pyruvate Kinases Activity in Vivo, in Vitro, in Silico
Lelevich Sergey Vladimirovich, Khrustalev Vladislav Victorovich, Barkovsky Eugene Victorovich, Shedogubova Tatyana Aleksandrovna

American Journal of Medical Sciences and Medicine
DOI: 10.12691/ajmsm-1-1-1

Clinical and Histological Features in Wegener Granulomatosis
Li Cavoli Gioacchino, Passantino Rita, Bono Luisa, Tortorici Calogera, Ferrantelli Angelo, Giammarresi Carlo, Rotolo Ugo

American Journal of Infectious Diseases and Microbiology
DOI: 10.12691/ajidm-1-1-2

The Curse of Lymphatic Filariasis: Would the Continual Use of Diethylcarbamazine Eliminate this Scourge in Papua New Guinea?
Anne Lanham, Lillian Mwanri

We have released issues of some journals on February 2013.
American Journal of Public Health Research
American Journal of Medical Sciences and Medicine
American Journal of Medical and Biological Research
American Journal of Pharmacological Sciences
American Journal of Clinical Medicine Research
American Journal of Nursing Research
American Journal of Cancer Prevention
American Journal of Medicine Studies
American Journal of Infectious Diseases and Microbiology
American Journal of Cardiovascular Disease Research
American Journal of Biomedical Research
American Journal of Mechanical Engineering
American Journal of Microbiological Research
Journal of Preventive Medicine
American Journal of Food and Nutrition
American Journal of Hypertension Research
American Journal of Sports Science and Medicine
Journal of Cancer Research and Treatment
American Journal of Zoological Research
Journal of Biomedical Engineering and Technology
American Journal of Microbiological Research
Biomedical Science and Engineering

We are pleased to announce that submissions are now being accepted for peer review for all the journals across all our subject areas in SciEP.

Please submit your manuscript via our online submission system:
http://www.sciepub.com/author/signin

To unsubscribe from our mailing list, please click on the following link: unsubscribe

I am looking forward to hearing from you.

Kind regards,

David Rain
Email:join@sciepub.com
Science and Education Publishing (http://www.sciepub.com)

Label_Title

Dear Label_Name,

Greetings! I hope you are well.

I am writing on behalf of an international publishing house, Science and Education Publishing, USA (http://www.sciepub.com).

Science and Education Publishing (SciEP) is a multidisciplinary open access publisher of Journals covering the fields of Science, Technology and Medicine. All SciEP publications are subject to high-quality peer review, editorial and production processes and are freely available online.

Welcome to the April, 2013 edition of SciEP newsletter, providing monthly summaries of articles on science & education related topics of interest to scientist and professionals. The mission of our journals is to provide an open access forum for publishing new findings on issues pertinent to scientific and academic field. We also invite you to submit original research papers, review articles, short reports, or case studies are considered for publication in our Multidisciplinary Journals.

American Journal of Public Health Research
DOI: 10.12691/ajphr-1-1-3

The Implementation of a Tele-Homecare System with Service Oriented Architecture and HL7 Message Transmission Standard
Ching-Sung Wang

American Journal of Medical and Biological Research
DOI: 10.12691/ajmbr-1-1-2

The Influence of Ethanol on Pyruvate Kinases Activity in Vivo, in Vitro, in Silico
Lelevich Sergey Vladimirovich, Khrustalev Vladislav Victorovich, Barkovsky Eugene Victorovich, Shedogubova Tatyana Aleksandrovna

American Journal of Medical Sciences and Medicine
DOI: 10.12691/ajmsm-1-1-1

Clinical and Histological Features in Wegener Granulomatosis
Li Cavoli Gioacchino, Passantino Rita, Bono Luisa, Tortorici Calogera, Ferrantelli Angelo, Giammarresi Carlo, Rotolo Ugo

American Journal of Infectious Diseases and Microbiology
DOI: 10.12691/ajidm-1-1-2

The Curse of Lymphatic Filariasis: Would the Continual Use of Diethylcarbamazine Eliminate this Scourge in Papua New Guinea?
Anne Lanham, Lillian Mwanri

We have released issues of some journals on February 2013.
American Journal of Public Health Research
American Journal of Medical Sciences and Medicine
American Journal of Medical and Biological Research
American Journal of Pharmacological Sciences
American Journal of Clinical Medicine Research
American Journal of Nursing Research
American Journal of Cancer Prevention
American Journal of Medicine Studies
American Journal of Infectious Diseases and Microbiology
American Journal of Cardiovascular Disease Research
American Journal of Biomedical Research
American Journal of Mechanical Engineering
American Journal of Microbiological Research
Journal of Preventive Medicine
American Journal of Food and Nutrition
American Journal of Hypertension Research
American Journal of Sports Science and Medicine
Journal of Cancer Research and Treatment
American Journal of Zoological Research
Journal of Biomedical Engineering and Technology
American Journal of Microbiological Research
Biomedical Science and Engineering

We are pleased to announce that submissions are now being accepted for peer review for all the journals across all our subject areas in SciEP.

Please submit your manuscript via our online submission system:
http://www.sciepub.com/author/signin

To unsubscribe from our mailing list, please click on the following link: unsubscribe

I am looking forward to hearing from you.

Kind regards,

David Rain
Email:join@sciepub.com
Science and Education Publishing (http://www.sciepub.com)

The Cytotoxic Effect of Aqueous Extract of Moringa Oleifera (Moringaceae) on Wistar Rats

Tchogou Atchadé Pascal, Semado Franck Médard, Agbogba Félicienne, Abissi Yollande, Kassa Roxane, Lokonon Jacques Ezéchiel, Medoatinsa Espérance, Houngbeme Alban, Anago Eugénie, Baba-Moussa Lamine, Gbenou Joachim Djimon, Senou Maximin*

Experimental and Clinical Biology Unit (UBEC), Medical and Pharmaceutical Biotechnology Research Laboratory (LaRBiMeP), National School of Applied Biosciences and Biotechnology of Dassa-Zoumé (ENSBBA), National University of Sciences, Technologies, Engineering and Mathematics of Abomey (UNSTIM), R Benin.

Abstract: Introduction: Moringa oleifera was a medicinal plant generally used by populations in the food as well as therapeutic field. Its use to treat anemia was observed in the Djougou area in northern Benin in traditional medicine. Although this plant underwent a lot of scientific study, there was still to our knowledge, a lack of scientific data on its effectiveness in the treatment of hemolytic type anemia. The aim of our study was to test its safety in vivo on Wistar rats. Methods: Wistar rats received by gavage a single dose of 2000 mg/kg of aqueous extract of Moringa oleifera leaves for the acute oral toxicity test (TOA). For the subchronic oral toxicity test (TSC), the Wistar rats received the aqueous extract by gavage at a daily dose of 200 mg/Kg of body weight per day for 28 days. The rats were weighed and blood samples were taken on day 0, then on day 14 for TOA and on day 28 for TSC, respectively. The renal assessment was carried out by the dosage of creatinine, the liver assessment by the transaminases AST and ALT and the immune assessment by the hemogram. The liver, kidneys and spleen were removed for histological analysis. The results were analyzed using the Mann Whitney test, the significance level being set at 5%. Results: The weight of the rats did not change significantly in the various acute or subchronic oral toxicity tests, suggesting the absence of physical disturbance in the rats. Serum creatinine did not change significantly, indicating preservation of renal function. It was the same for AST and ALT transaminases, indicating an absence of hepatic cytolysis. Hepatic, renal and splenic parenchyma showed no atypia. Conclusion: The aqueous extract of Moringa oleifera leaves did not reveal any toxic effect in the acute or subchronic state and offered prospects for its use in the treatment of anemia.

Keywords: Moringa oleifera, oral toxicity, subchronic liver, kidney and spleen toxicity

Received September 11, 2023; Revised October 13, 2023; Accepted October 20, 2023

Citation: Tchogou Atchadé Pascal, Semado Franck Médard, Agbogba Félicienne, Abissi Yollande, Kassa Roxane, Lokonon Jacques Ezéchiel, Medoatinsa Espérance, Houngbeme Alban, Anago Eugénie, Baba-Moussa Lamine, Gbenou Joachim Djimon, Senou Maximin, The Cytotoxic Effect of Aqueous Extract of Moringa Oleifera (Moringaceae) on Wistar Rats. American Journal of Medical Sciences and Medicine, 11 (3). 92-97. January 2023. doi:10.12691/ajmsm-11-3-5

1. Introduction

Anemia was defined by the fall in the level of hemoglobin in the circulating blood compared to normal values [1]. It was a syndrome observed in various pathologies such as genetic anomalies, infections [2]. The World Health Organization (WHO) estimated that 1.62 billion people suffer from anemia, i.e. a global prevalence of 24.8% with a predominance in Africa and South-East Asia, i.e. 67 respectively. 7% and 65.5% [3]. In Benin, 72% of children aged 6-59 months suffered from anemia, of which 3% in its severe form and 41% in its moderate form, then 58% of women aged 15 to 49 suffered from anemia: 26% in its mild form, 30% in its moderate form and 2% in its severe form [4]. Treatment varied depending on the type. This could be iron, vitamin B12 or B9 [5-6]. Given the high cost of treatment, the inaccessibility and adverse effects of conventional synthetic drugs used in developing countries, the search for alternative and/or complementary strategies was necessary for the use of synthetic antianaemics and other conventional drugs. WHO estimated that 80% of rural populations in developing countries depend on traditional medicine for primary health care [7], including anemia and its complications. In Benin, out of the 30,700 species of plants inventoried in forest ecosystems [8] which were used by local populations as food plants and others as medicinal plants, including Moringa oleifera and Jatropha Gossypiifolia [8]. Moringa oleifera leaf powder contained most of the essential nutrients needed for good health [9-12]. The leaf powder was rich in multiple minerals and vitamins, including iron, vitamin A (carotenoid) and vitamin C, which were important for iron metabolism. Additionally, Moringa had an added benefit for solving multiple malnutrition problems because it was rich in all essential amino acids, which were building blocks of proteins necessary for cell growth [2,13] [15,16]. Studies conducted in different countries like Senegal and India, the use of Moringa has been reported to reduce malnutrition in children as well as vitamin A and protein deficiencies [13,17] [18,19,20] without any risk of toxicity [21,22,23] and no adverse side effects [18]. The nutritional potential of Moringa leaf powder maked it an important ingredient for improving nutrient diversification in complementary foods for children [13]. In the rich and diversified flora of Benin, Moringa oleifera and many other plants were used in traditional medicine to treat anemia. These plants, although having already been the subject of several scientific studies on their therapeutic virtue; have not yet been proven to our knowledge in Benin, their effectiveness on hemolytic anemia. It is for this reason that this study was undertaken with the aim of evaluating the cytotoxic effect of the aqueous extract of Moringa oleifera (Moringaceae) on Wistar rats. Such an evaluation would allow not only their rational use but also their possible transformation into improved traditional medicines (MTA) to be used without toxic risk by poor populations.

2. Materials and Methods

Animal material

The animal material used in this study consisted of albino Wistar strain rats aged 4 to 5 months and with an average body weight of approximately 210 g. These rats were acclimatized to ambient conditions at a constant temperature of 25 ± 1°C in the animal facility of the Experimental and Clinical Biology Unit (UBEC), of the National School of Applied Biosciences and Biotechnologies (ENSBBA) of Dassa, National University of Sciences, Technologies, Engineering and Mathematics (UNSTIM) in Benin. The rearing took place in a well-ventilated room with a cycle of 12 hours of light and 12 hours of darkness. The rats were kept in wire mesh cages with feeders and drinkers, and had free access to drinking water and food. They were supplied with drinking tap water and their daily diet consisted of a mixture of granulated feed presented in the form of kibble and marketed by Véto Services (Benin). The breeding enclosure was regularly cleaned to guarantee optimal development of the animals protected from any infection.

Identification and preparation of plants

The samples studied were fresh leaves collected in the northern Benin region, specifically in Djougou in the department of Donga and brought back to the laboratory for drying. The samples were spread out in a cold drying room (22°C) for about 14 days, after which time the samples were brittle and were practically anhydrous. The identification was carried out at the National Herbarium of Benin. The extraction of the total chemical principles was done using the method of maceration in accordance with the traditional use of plants. Based on the extraction techniques cited in the literature [24], 50 g of powder were dissolved in 500 mL of distilled water. The mixture was left stirring continuously for 48 hours. After cooling, the mixture obtained was filtered (3 times in a row) on absorbent cotton and the filtrate was transferred to a 1000 mL flask then subjected to evaporation until dryness at 40° C. using a rotavapor (Heidolph Laborota 4000 efficient) coupled to a water chiller (Julabo FL 300). The dry residue obtained represented the maceration.

Larval toxicity test

Larval cytotoxicity was commonly performed using shrimp larvae in seawater (Artemia salina) in the presence of the test solution. Its interest lied in the knowledge of the safety of use of the plants whose extracts were tested. Indeed, there was a correlation between toxicity on shrimp larvae and cytotoxicity on 9PS and 9KB cells (human nasopharygeal carcinoma) on the one hand, lung carcinoma A-549 cells and carcinoma HT-29 cells. colon on the other hand. The test, used successfully in 2010 by Fatondji et al., in 2012 by Sakirigui et al., and in 2014 by Houngbeme et al., was carried out according to the method of Michael et al., taken over by Vanhaecke et al., then by Sleet & Brendel. Artemia salina eggs were incubated in seawater until young larvae hatch (48 hours). A stock solution of the extract was prepared by dissolving 200 mg of extract in 4 mL of distilled water; i.e. a concentration by weight of 50 mg/mL. Ten [11] successive half (1/2) dilutions of the stock solution with seawater were carried out. A defined number of larvae [17] was introduced into each dilution.

All the solutions as well as control solutions containing no active substance were left under stirring for 24 hours. Macroscopic counting of the number of surviving larvae in each solution maked it possible to assess toxicity. In the case where deaths were observed in the control medium, the data were corrected by Abbott's formula: %death = [(test – control) / control)] x 100. The dose-response data were transformed by logarithm and the LC50 is thus determined by linear regression.

To assess the degree of toxicity from the LC50 values, we used the correspondence table (Table 1) drawn up by Mousseux in 1995 and widely used in the literature [24].

Table 1. Correspondence between LC50 and toxicity

Assessment of acute and subchronic oral toxicity

The Toxicity Test was performed according to the recommendations of the modified Organization for Economic Cooperation and Development (OECD) Guideline 423 at doses of 200, 300, 1200, 2000 and 5000 for testing chemicals [25]. Three (03) animals were used for the test. Their weights were determined before and after the test. Hepatic and renal functions were assessed at the beginning and at the end of the experiment on blood samples (assay of transaminases and serum creatinine) and histological sections of the kidney, liver and spleen (search for cell lesions).

Acute toxicity

It consisted of testing Moringa oleifera extract at a single dose of 2000 mg/kg on rats. The test was carried out on 6 Wistar rats. The behavior of the rats was observed as well as the number of deaths over a period of 14 days. After 15 h of fasting, they were distributed as follows: control group consisting of 3 rats receiving distilled water, at a rate of 10 ml/kg; experimental batch consisting of 3 rats, receiving the extract of Moringa oleifera, at a rate of 2000 mg/kg. A behavioral observation was carried out 3 hours after administration of the extracts. Then hydration and feeding were performed daily for 14 days. During this period, signs of toxicity including coat change, motility, tremors, stool appearance, mobility and death were monitored. At the end of the treatment, the rats were fasted for 24 h, then a blood sample was taken, followed by a dissection after general anesthesia with ether. The organs removed were the liver, kidneys and spleen. The latter were rinsed with a 0.9% saline solution, then placed in 10% diluted formalin for the histological study.

Subchronic toxicity

It was carried out on 6 Wistar rats divided into 3 equal groups of 3 rats each as follows: group 1, receiving distilled water at a rate of 1 ml/100 mg of body weight (control group); batch 2 (experimental batch) receiving a solution of Moringa oleifera extract at 200 mg/kg body weight every day for 28 days. The rats were fed and hydrated ad libitum. At the end of the treatment, the rats were fasted for 24 h, then a blood sample was taken, followed by a dissection after general anesthesia with ether. The organs removed were the liver, kidneys and spleen. The latter were rinsed with a 0.9% saline solution, then placed in 10% diluted formalin for the histological study.

Blood tests

Serum creatinine was measured to explore renal function. AST and ALT transaminases were measured for liver function. The number of white blood cells was measured as hematological parameters.

Histological studies

A standard historical study was carried out for the acute and subchronic oral toxicity test on the organs removed after sacrifice of the different groups of animals. Kidneys, liver and spleen were removed, fixed in a 10% buffered formalin solution and embedded in paraffin. Sample sections (5 μm) were mounted on glass slides, deparaffinized and hydrated. For histological analysis, the sections were stained with hematoxylin and eosin (H&E), according to a standard protocol [18]. Pictures were taken at 400X magnification.

Statistical analyzes

Parameter means were compared using the Mann Whitney test. The significance level was set at 5%.

3. Results

In vitro Cytotoxicity of the extract

The sensitivity curve of the Artemia salina larvae to the extract was plotted from the numbers of dead larvae recorded after reading the test, as a function of the concentration of the extract. The curve was represented by the figure below.

The analysis of the LC50 values of the extracts tested by the correspondence table (Table 1), allowed to say that the tested extract did not present any toxicity in the range of concentrations analyzed. In fact, the LC50 values obtained were above the toxicity limit set (0.1 mg/mL) Figure 1.

Figure 1. Sensitivity curve

The aqueous extract of Moringa oleifera leaves showed no acute oral toxicity. The results of the acute oral toxicity test of the extract were presented in (Table 2).

On D0, the weight of the rats was 147 ± 9 g, the serum creatinine was 9 ± 1 mg/L, the AST and ALT transaminases were respectively 101 ± 12 U/L and 73 ± 11 U/L, the mean number of blood leukocytes was 10.1 ± 1.4 G/L. These different parameters did not change significantly on D14, indicating an absence of physical disturbance, an absence of alteration of the renal, hepatic and immune functions of the rats. This result suggested an absence of toxicity of the extract in the acute state (Table 2).

Table 2. Acute oral toxicity of Moringa oleifera extract

The aqueous extract of Moringa oleifera leaves showed no subchronic toxicity. The results of subchronic oral toxicity test of the extract were presented in (Table 3).

On D0, the weight of the rats was 185 ± 29 g, the serum creatinine was 9 ± 2 mg/L, the AST and ALT transaminases were respectively 91 ± 13 U/L and 53 ± 12 U/L, the mean number of blood leukocytes was 13.3 ± 2.1 G/L. These different parameters did not change significantly on D28, indicating an absence of physical disturbance, an absence of alteration of the renal, hepatic and immune functions of the rats. This result suggested an absence of toxicity of the extract in the subchronic state (Table 3).

Table 3. Subchronic oral toxicity of Moringa oleifera extracts

Moringa oleifera leaf aqueous extract did not change liver parenchyma

Figure 2 showed the renal paremchyma. In acute (AOT) and subchronic (SCT) oral toxicity tests, the liver of rats did not show atypia with a normal parenchyma like that of control rats. The hepatocytes (arrows) were organized into laminae separated by conspicuous venous sinusoids (VS) which communicate with the centrilobular vein (CV) Figure 2.

Figure 2. Liver histology in acute and subchronic oral toxicity tests of aqueous Moringa oleifera extract (400X magnification)

Moringa oleifera leaf aqueous extract did not change renal parenchyma

Figure 3 showed the renal parenchyma. In the acute (AOT) and subchronic (SCT) oral toxicity tests, the renal parenchyma did not show any atypia and was like that of the control rats. The glomeruli (G), the proximal tubes (PT) and the distal tubes (DT) were clearly visible Figure 3.

Figure 3. Renal histology in acute and subchronic oral toxicity tests of aqueous Moringa oleifera extract (400X magnification)

Moringa oleifera leaf aqueous extract did not change splenic parenchyma

Figure 4 showed the splenic parenchyma. In acute (AOT) and subchronic (SCT) oral toxicity tests, the splenic parenchyma was typical as in control rats. The central arteries (CA) with their periarteriolar sheaths (PS) and germinal centers (GC) were clearly visible in the white pulp. The venous sinusoids (VS) were well separated by the Billroth cords (BC) in the red pulp Figure 4.

Figure 4. Histology of the spleen in acute and subchronic oral toxicity tests of aqueous Moringa oleifera extract (400X magnification)

4. Discussion

This study focused on the evaluation of the cytotoxic effect of the aqueous extract of Moringa oleifera (Moringaceae) on rats Wistar. That was a plant with anti-anemic property whose leaves could be used for the prevention and correction of malnutrition in because of its exceptional nutritional qualities [26-32]. The in vitro toxicity study using the larval toxicity test did not show any toxic effect. This result is comparable to that obtained with the aqueous extract of Cocos nucifera L. (Arecaceae) root, also used to treat anemia [41]. In the acute or subchronic in vivo toxicity study, the behavior and weight of the rats were not modified. Similar results were obtained by Fikriansyah et al [32] during their study on the cardioprotective effect of Moringa oleifera but by injecting a dose of 4.67 mg/kg. However, these results were contrary to those obtained by Sudha P. et al [33] who reported increased mortality of rats tested at a dose of 2000 mg/kg of body weight, prompting them to opt for doses of 750 mg/ kg and 250 mg/kg in their evaluation tests. In addition, Lemita N. et al [34] reported an increase in the weight of rats after administration of Moringa Oleifera leaf extract to rats at a dose of 150 mg/kg by the intraperitoneal route. In the acute or subchronic oral toxicity tests of the extract, the ALAT and ASAT transaminases were not modified. These observations suggested that the aqueous extract of Moringa Oleifera would have hepatoprotective effects. These results were similar to those reported by Senou et al [35], who noted a non-significant decrease in ALT enzyme levels after subchronic administration of an aqueous extract of Senna alata at a dose of 200 mg/kg in the treated rats. In a similar study, Luka et al [36] explained that a decrease in ASAT and ALAT hepatic enzymes could indicate a hepatoprotective effect of the crude ethanolic extract of the roots of Diospyros mespiliformis hochst (Ebenaceae). In a study carried out in diabetic rats, Lemita N. et al [34] observed this hepatoprotective effect when they noted a very significant increase in ALT and AST transaminases in untreated diabetic rats compared to controls, and compared to diabetic rats treated with Moringa Oleifera. This result was confirmed by a liver histology that remained normal, an observation also noted by [34] and [35] in their respective study. Evaluation of acute and subchronic toxicity of Moringa oleifera extracts in rats showed that creatinine did not experience a significant increase, suggesting that the aqueous extract of Moringa oleifera leaves was not nephrotoxic. This observation was confirmed by histology which showed that the renal parenchyma with glomeruli, proximal and distal tubules and collecting tubules were normal. These results were similar to those reported by Lemita N. et al [34] in diabetic rats treated with Moringa oleifera compared to untreated rats, noting a total absence of lesions in all the organs studied, including the kidneys in the diabetic rats treated with Moringa Oleifera. These results were also observed by other authors [37] on the bark of Psorospermum febrifugum, an anti-anemic plant. An absence of subchronic oral toxicity of the flowers of Senna alata, an antiamemic plant, was also observed by authors [35] and [38], on the liver, kidneys and heart of Rat Wistar. In acute and subchronic oral toxicity tests, immune function as assessed by white blood cell count was not affected. The histology of the spleen, a peripheral immune organ, was not altered, confirming the non-toxic nature of the plant. Similarly, similar observations have been made with the aqueous extract of the root bark of Cocos nucifera, an anti-anaemic plant [40].

5. Conclusion

The different results obtained on the biochemical, hematological and histological level during in vitro and in vivo toxicity tests, reassure on the safe use of Moringa oleifera leaves in the perspective of treatment of anemia. Nevertheless, it would be important to continue the evaluation of the toxicity of the leaves of this plant in the long term with chronic toxicity tests. This would make it possible to envisage the formulation of these extracts in improved traditional medicine and accessible to the rural populations who were the most numerous and the most exposed.

Statement of Competing Interests

The authors declared no conflicts of interest.

References

[1]   | Bertrand G. Diagnostic des anémies hémolytiques, CHU Henri Mondor Créteil Paris, 2011.
[2]   | Assobayire FS; Adou P, Davidson L, Cook JD Hurrel R. Pevalence of iron deficiency with and without. Concurrent anemia in population group with high prevalences of malaria and other infections: a study in Cote d’iviore. Am J Clin Nutr; 2001, 74:776-82.
[3]   | OMS. https://www.who.int/fr/health-topics/anaemia, 2020, consulté le 24 juillet 2023.
[4]   | OMS (Genève). Prevalence of anemia 1993-2005: who global Database on anemia. Genève: OMS : 2008, 25.
[5]   | INSAE and ICF. Enquête démographique et de sante.´ https://www.insae-bj.org/images/docs/insae, statistiques/enquetesrecensements/EDS/2017-2018/1.Benin EDSBV Rapport final.pdf Accessed: 2020-05-15.
[6]   | Movaffaghi Z, Hasanpoor M. Effect of therapeutic touch on blood hemoglobin and hematocrit level JHolist Nurs 2006; 24 :41-8.
[7]   | Seidou A. Evaluation de quelques proprietes de « Forte » utilisée pour traiter l’anemie, these de doctoraten pharmacie de la Faculté des Sciences de Santé ; FSS/UAC, 2013, Bénin 100p.
[8]   | OMS (Harare). Promotion du role de la médécine traditionelle dans le système de santé : stratégie de la région africaine AFR/ RC / 9.Harare : bureau régional de l’Afrique : 2001, 20.
[9]   | Akoegninou et al. flore analytique du Bénin, 2006.
[10]   | Angela et al. Proposal of an Adapted Physical Activity Exercise, Protocol for Women with Osteoporosis-Related Vertebral Fractures: A Pilot Study to Evaluate Feasibility, Safety, and Effectiveness, 2009.
[11]   | Ashifaq et al. Agric. including coastal areas. It is a fast-growing, drought- resistant tree. Soc. Sci., Vol. 8, No. 3, 2012. 116.
[12]   | Hiawatha Bey. All things Moringa,p. 4,5, 2010. (4) Carter, M.R., and E.G. Gregorich (eds). 2008. Soil sampling and methods of analysis. 2nd, 2010.
[13]   | Yang et al. Moringa and other highly nutritious plant resources: Strategies, standards and markets for a better impact on nutrition in Africa. Accra, Ghana, November 16-18, 2006.
[14]   | Angela E. Shija. Effect of Moringa Oleifera leaf powder supplementation on reducing anemia in children below two years in Kisarawe District, Tanzania, 2019.
[15]   | Ashfaq, M., S.M.A. Basra and U. Ashfaq. Moringa: A Miracle Plant of Agro-forestry. J. Agric. Soc. Sci., 2012, 8: 115–122.
[16]   | Hiawatha Bey. All Things Moringa: The Story of an Amazing Tree of Life, 2010, 40p.
[17]   | Yang et al. Both IL-12p70 and IL-23 Are Synthesized during Active Crohn's Disease and Are Down-regulated by Treatment with Anti-IL-12 p40 Monoclonal Antibody, 2006.
[18]   | Fahey. Moringa oleifera: A Review of the Medical Evidence for Its Nutritional, Therapeutic, and Prophylactic Properties. 2005, Part 1.
[19]   | Fuglie. Nutritional evaluation of complementary food formulations from maize, soybean and peanut fortified with Moringa oleifera leaf powder was carried out, 2001.
[20]   | Mahmood, Mugal, & Haq. Moringa oleifera: a natural gift-A review. Mahmood, Khawaja Tahir; Mugal, Tahira; Haq, Ikram Ul. Journal of Pharmaceutical Sciences and Research, 2010.
[21]   | Srikanth. Non-traditional security threats in the 21st, 2014.
[22]   | Stohs& Hartman. Review of the Safety and Efficacy of Moringa oleifera. Phytother Res. 2015 Jun; 29(6):796-804.
[23]   | Luqman, Srivastava, Kumar, Maurya, & Chanda, Experimental Assessment of Moringa oleifera Leaf and Fruit for Its Antistress, Antioxidant, and Scavenging Potential Using In Vitro and In Vivo Assays, 2012.
[24]   | Devaraj, Asad, & Prasad. Effect of leaves and fruits of Moringa oleifera on gastric and duodenal ulcers, 2007. Devaraj, V.C.; Asad, M.; Prasad, S.

[25]  
Houngbeme AG, Gandonou C, Yehouenou B, Kpoviessi SDS, SohounhloueD, Moudachirou M and Gbaguidi FA. Phytochemical analysis, toxicity and antibacterial activity of Benin medicinal plants extracts used in the treatment of sexually transmitted infections associated with HIV-AIDS. Int J Pharm Sci Res; 2014, 5 (5): 1739-1745.

[26]   | Organisation de Coopérat ion ET de Devellopement Economique, OCDE Bookshop. Essai N0423: Toxicité orale aigue-Methode par classe de casassus toxicité. (End ligne) (consulté le 15 Juin 2021), 2020, URL http://WWW.oecd.org/dataoecd/17/50/1948370. pdf.
[27]   | Rosidah, Yam, M.F., Sadikun, A., Ahmad, M., Akowuah, G.A. and Asmawi, M.Z. Toxicology Evaluation of Standardized Methanol Extract of Gynura procumbens. Journal of Ethnopha Macology, 2009, 123, 244-249.
[28]   | Abdou S. N. Valorisation des feuilles de Moringa oleifera Lam en tant que produits phytotherapeutiques, mémoire de master, Univsersité de Mahajanga, Mahajanga, and 53p, 2006.
[29]   | Chourouk B., Rayane L. Propriétés nutritionnelle ET fonctionnelle de la plante Moringa oleifera, mémoire de master, Université Mohamed Khider de Biskra, 60p, 2022.
[30]   | BleuG.M. AhuiB.M.L.B., KonanB.A. Brou A.A.E.F.,Obou C.O. & TraoréF. Etude des Effets Antianémiques d’un Complément Alimentaire à Base de Feuilles de Jatropha gossypiifolia chez des Rats Wistar.ESI Preprints.
[31]   | Stohs S. J., Hartman M. J. Review of the Safety and Efficacy of Moringa oleifera, Phytother. Res. 2015, 29: 796–804.
[32]   | Magagi S., Oumarou D. H., Ibro G., Kaka S., Balla A. Perceptions endogènes ET utilisations des feuilles de Moringa oleifera en milieu rural de la République du Niger: cas des régions de Tillabéri et de Maradi, Int. J. Biol. Chem. Sci. 2022, 16(5): 2070-2087.
[33]   | Fikriansyah, F., Widiastuti, M., Wulandari, N., Tirtanirmala, P., & Murwanti, R. Cardioprotective effect of Kelor (Moringa oleifera) leaf ethanolic extract against doxorubicin induced cardiotoxicity in rats. Indonesian Journal of Cancer Chemoprevention, 2015, 6(2), 53-57.
[34]   | Sudha P., Syed M. B. A., Sunil S. D. and Gowda K. C. Immunomodulatory activity of methanolic leaf extract of moringa oleifera in animals, Indian J Physiol Pharmacol 2010; 54 (2): 133–140.
[35]   | Lemita N. Effet cytoprotecteur de Moringa Oleifera sur le diabète expérimental chez les rats Wistar, mémoire de master, Université de Larbi Tebessi –Tébessa, Algérie, 2020.

[36]  
Senou, M., Dehou, R., Agbogba, F., Tchogou, P., Abissi, Y., Houngbeme, A., Kpossou, G., Anago, E. and Attakpa, E. Safety of the Aqueous Extract of the Leaves of Senna alata (L.) Roxb. (Leguminosae-Caesalpinioideae), a Plant Used in Benin to Treat Infections. Journal of Biosciences and Medicines, 2022, 10, 86-95.

[37]  
Luka, J., Badau, S.J., Mbaya, A.W., Gadzama, J.J. and Kumshe, H.A. Acute Toxicity Study and Effect of Prolonged Administration (28 Days) of Crude Ethanolic Root Extract of Diospyros mespiliformis Hochst (Ebenaceae) on Clinical, Haematological and Biochemical Parameters of Albino Rats. Journal of Ethno pharmacology, 2014, 153, 268-273.

[38]  
Agbogba, F., Senou, M., Tchogou, A.P., Lokonon, J.E., Sacramento, T.I., Medoatinsa, E., Kanfon, R.E., takpa, E., Dougnon, T.V., Agbangnan, D.C.P., Loko, F., Laleye, A., Ategbo, J.M., Baba-Moussa, S.L., Agbonon, A. and Sezan, A. Ethyl Acetate Fraction of Psorospermum febrifugum Spach Aqueous Extract Did Not Exhibit Acute or Sub-Chronic Toxicity. Experimental Study on Wistar rats. International Journal of Biology, 2019, 11, 123-129.

[39]   | Ogbe, R.J., Adoga, G.I. and Abu, A.H. Antianemic Potentials of Some Plant Extracts on Phenyl Hydrazine-Induced Anemia in Rabbits. Journal of Medicinal Plants Research, 2010, 4, 680-684.
[40]   | Owolabi J. O., Ogunsola O. A., Fabiyi O. S. Histological Assessment of Moringa Oleifera Ameliorative Activities on Lead Toxicity in the Spleen of Adult Wistar Rats. World Journal of Life Sciences and Medical Research; 2014, 3(2):63-6.
[41]   | Tchogou, A.P., Senou, M., Agbangnan-Dossa, C.P., Agossadou, A., Assogba, F., Dougnon, T.V., Klotoe, J.R., Gbenou, J., Sezan, A., Laleye, A. and Loko, F. Test of the Safety of Cocos nucifera L. (Arecaceae) Root Aqueous Extract. Journal of Chemical, Biological and Physical Sciences, 2017, 7, 282-291.
[42]   | Mukinda, J.T. and Syce, J.A.Acute and Chronic Toxicity of the Aqueous Extract of Artemisia afra in Rodents. Journal of Ethnopharmacology, 2007, 112, 138-144.