Phytochemical Screening, Antioxidant Activity, and Acute Toxicity Evaluation of Senna italica Extract Used in Traditional Medicine

Medicinal plants such as Senna italica are increasingly used for their purgative virtues to treat stomach aches, fever, and jaundice. This study aims to screen the phytochemical compounds and to assess the antioxidant activity in vitro and the acute oral toxicity in vivo of Senna italica leaves. The plant was harvested, dried, pulverized, and preserved. Phytochemical screening was performed using different laboratory protocols. Ethanolic and aqueous extracts were, respectively, obtained by maceration and decoction technics. The assay for free radical scavenging was used to examine the antioxidant activity using DPPH. Acute oral toxicity was performed with aqueous and ethanolic extracts at 5000 mg/kg of body weight on female albinos Wistar rats, weighing 152.44 ± 3.68 g. Subjects were checked for any signs of mortality and macroscopy toxicity during the 14 days of the study. Biochemical and hematological parameters were measured to assess liver and kidney functions, and histological analysis of these organs was conducted. Phytochemical analysis highlighted the presence of total phenols, flavones, tannins, alkaloids, and quinone derivatives. Semiethanolic (78 μg/mL), ethanolic (9.7 μg/mL), and aqueous extract (9.2 μg/mL) showed an interesting antioxidant activity. Biochemical and hematological parameters were normal and not significantly different (p > 0.05). The plant extracts did not produce any toxic effect or mortality at the provided dose. Senna italica extracts induced an increase in the volume of liver and kidney tissues but no necrosis. Thus, lethal dose 50 of Senna italica leaf extract is probably higher than 5000 mg/kg.


Introduction
According to the World Health Organization, various plant fractions and their dynamic components are used as traditional medicines by 80% of the global population [1]. One of these plants is Senna italica, colloquially known as agouema, an herbaceous plant or small deciduous shrub reaching 60 cm high generally with prostrate stems. Its leaves are organized in a spiral as encountered in Leguminosae-Caesalpinioideae family. Tey are glaucous, glabrous, alternate, and paripinnate with 5 to 6 pairs of obliquely oblong, obovate leafets, and adnate at both ends with pointed apices [2]; the leaves, the ripe seeds, and the pods of Senna italica (S.italica) have always been consumed for their purgative virtues. Taken as a maceration or decoction, they may be useful to treat jaundice, stomach aches, venereal diseases, fever, and bilious crises as well as intestinal afections. Te leaves are dried, pulverized, and used as a remedy for skin troubles such as ulcers and burns. Te fowers are employed to purge and induce childbirth. Te macerated roots are taken to treat colics and fu. Te macerated roots are used to treat colic and fu. Boiled roots are used to dress wounds [3]. Te infusion is smeared on eyes to relieve sores. S. italica roots are also a constituent of various medicines taken against indigestion, liver problems, spleen disorders, dysmenorrhea, vomiting, and nausea. In Malawi, the root infusion is used to treat infantile diarrhea. Te value of Senna italica. as a grazing plant is not unanimous [4]. In East Africa, most domestic animals eat it, while in West Africa, they seem to avoid it. On top of the purgative efects of the mature seeds, the young seeds are consumed as an appetizer or a vegetable in the region of Sahel. In Mauritania, the seeds of Senna italica Mill. are smoked. Sold as "neutral henna" or "blond henna," the leaves are also exploited as conditioner to make hair shiny. It can give hair a yellowish tint [5].
Several anthraquinones such as aloe emodin, chrysophanol, rhein, sennosides, and sennidines have been isolated from the leaves and pods of Senna italica. Tese diferent compounds are responsible for the purgative efect of this plant. Chrysophanol is another active component of "neutral henna." Te proportion in the leaves varies from 1.1 to 3.8% of dry weight. Te pods show a much lower content than those of the leaves. In addition, the leaves contain steroids (α-amyrin, β-sitosterol, and stigmasterol) and favonoids (kaempferol, quercetin, and apigenin) [3]. Te ethanolic extract of S. italica plant possesses some interesting antipyretic and antiinfammatory properties. 1,5-dihydroxy-3-methoxy-7-methylanthraquinone obtained from Senna italica Mill. can be used against several Gram-negative and Gram-positive bacteria. It also shows an anticarcinogenic activity in vitro [5]. Toxicity assays performed on rabbits and goats fed with the plant foliage were negative. Chicks and rats nourished with 10% of seed diet presented symptoms of toxicity, but no mortality sign during the 6-week trial. A 2% seed diet stimulated chick growth. Seeds produce a gum soluble in water and mainly composed of Dgalactose and D-mannose [6,7].
Te ethanolic and aqueous extracts of S. italica leaves have free radical scavenging, antioxidant, and secondary metabolite efects and are nontoxic for rats.
To explore the phytochemical constituents of Senna italica, we performed phytochemical screening and determined the phenolic composition. To assess the extracts capacity for free radical scavenging, we measured their antiradical activity. To check if the extracts are toxic or not for rats, we executed the acute toxicity test on Wistar rats following the OECD protocol.

Medicinal Plant
Extracts. An extraction of S. italica leaves was conducted using traditional medicine techniques, utilizing aqueous, ethanolic, and semiethanolic methods.
Te leaves were freshly collected from the Grand Popo (Agoué) commune in the south of Benin and identifed in the Benin National Herbarium under the identifcation number N°YH 722/HNB. Following collection, the leaves were dried at a controlled temperature of 20 to 25°C, ground into powder, and stored in a hermetically sealed container using a proper protocol [8]. For the aqueous extract, 50 grams of S. italica leaf powder was boiled in 500 mL of water at 100°C. Te ethanolic extracts were prepared by mixing 50 grams of leaf powder with ethanol or 50% ethanol and continuously agitating the mixture for 72 hours. Te resulting macerate was fltered using the Whitman paper N°1, and the fltrates were concentrated in a rotavapor before being dried in a proofer at 50°C. Te extraction yield was calculated using the following formula:

Determination of Total Phenolic, Tannins, and Flavonoid
Contents. Te total phenolic compounds in the extracts obtained from 1/15 sample (w: v) of fresh nuts were measured using Folin-Ciocalteu's method, as described by singleton, six solvents (water, methanol, ethanol, water-HCl 1%, ethanol-HCl 1%, and methanol-HCl 1%) were used to determine the extraction capacity of C. nitida. Te nuts were ground with a grinder, adapted to 96 well-plates, and 25 μl of Folin-Ciocalteu's reagent (50% v/v) were combined with 10 μl of 1 mg/ml (w/v) of the nuts extract. After incubation at room temperature for 5 min, 25 μl of 20% (w/v) sodium carbonate (Na 2 CO 3 ) and water were added to reach a volume of 200 μl per well. Blanks were prepared using water instead of the reagent to minimize the impact of interfering compounds. Te absorbance was measured at 760 nm after incubation (30 min) using a multiwell plate reader. Te assays were performed in triplicate at least, and the results were expressed as microgram gallic acid equivalent per 100 grams of extract using gallic acid (0-500 μg/ml) as a standard. Te total favonoids in each sample were quantifed using the aluminum trichloride method adapted to 96 well-plates [10]. Hundred microliters of methanolic AlCl3 (2%) were mixed with 100 μl of appropriate dilution of the extract solution. After incubation (15 min), the absorbance was measured at 415 nm using a multiplate Epoch spectrophotometer Biotech connected to a computer with the help of "Gen5" software against a blank (mixture of 100 μl methanolic extract solution and 100 μl methanol) and compared to a quercetin (0-50 μg/ml) calibration curve (R 2 � 0.99). Te favonoid content was expressed as mg of quercetin substitutes per 100 g of extract.
Te condensed tannin content was determined using the vanillin assay recommended by Belyagoubi et al. [11]. 1500 μL of vanillin/methanol solution (4%, w/v) was added to 50 μL of extract (S1 or S2) and followed by the addition of 750 μL of 37% HCl. Te sample was incubated at room temperature for 20 min, and the absorbance was measured at 550 nm against a blank. Catechin was used as a standard for calibration curve, and the total proportion of condensed tannins was calculated as mg of catechin equivalents per g of dry matter (mg CE/g DM).

Antioxidant Assay.
Te antioxidant assay of diferent extracts was evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH.) method, which was often used for its simplicity. It is a technic based on the reduction of an alcoholic solution of DPPH. Provided an antioxidant that gives hydrogen or a proton, the nonradical form DPPH-H was formed [12,13]. Te decoloration of DPPH radical depends on the concentration of the diferent extracts used. Te extract-free radical scavenging activity, expressed as IC50, defnes the efective concentration of the substrate that causes loss of 50% of DPPH radical activity [14,15]. Te optic densities were measured at 517 nm and used to calculate the percentage of scavenging of the DPPH radical, which was proportional to the antiradical power of the sample [16]. Tese IC50 are determined from curves that estimate the antioxidant activity of extract as a percentage of crude extract concentration [12]. A volume of 100 μL of each extract at diferent concentrations was added to 1900 μL of the ethanolic solution of DPPH (40 μg/mL). Te negative control was prepared in parallel by mixing 100 μL of extraction solvent with 1900 μL of DPPH solution. After incubation at room temperature and in the darkness for one hour, absorbances are taken at 517 nm using a HACH LANGE DR 3900 spectrophotometer.
Te percentage of trapping was calculated by the formula:  [17]. Te temperature of the room was maintained at 25 degrees Celsius, and artifcial lighting was provided in alternating cycles of 12 hours of light and 12 hours of darkness. Te rats were provided with standard granulated food from a commercial food supplier in Benin and were given access to drilling water ad libitum.

Experimental Design.
Te acute toxicity study was conducted in compliance with the OECD Guidelines (Organization for Economic Cooperation and Development, Guideline-423, adopted on 17 December 2001). Senna italica extract was administered once to the rats as a single dose of 5000 mg per kg of body weight in accordance with the acute oral toxicity protocol of the OECD Guidelines for Chemicals Testing 423, adopted December 17, 2001, and in accordance with good laboratory practice. Te rats were divided into three groups of three rats each: the frst group of control rats, the second group of normal rats which received the aqueous extract of Senna italica via tube-feeding at a dose of 5000 mg/ kg body weight, and fnally, the third group of normal rats which received Senna italica ethanolic extract via tubefeeding at a dose of 5000 mg/kg body weight. Te animals were fasted for 12 hours prior to the administration of the extracts, after which they were weighed and then the extracts were given based on their fasting body weight. Te rats were observed for 14 days during the experiment to monitor their behavior. Te extracts were considered toxic if 50% of the experimental rats had died.

Biochemical Analysis.
After allowing blood samples in non-EDTA coated tubes to clot for 5 minutes, they were immediately centrifuged at 3000 rpm for 10 minutes to separate the serum for analysis. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, alkaline phosphatase, gamma GT, and total cholesterol were analyzed using a chemistry analyzer (Hu-mastar 200, Germany). Electrolytes were analyzed using an electrolyte analyzer (Humate plus 5, Germany).

Histopathological Analysis.
On the 14th day of the experiment, the rats were sacrifced with thiopental (30 mg/ kg) after sampling. Te kidney and liver were removed, fxed in a 10% formalin solution, and parafn-embedded. Microtome sections of approximately 3 to 5 μm were made and mounted on glass slides. Te sections were dewaxed in toluene and hydrated in decreasing alcohol baths. For histological analysis, the sections were stained using hematoxylin and eosin (H and E) following the standard protocol [18].

Ethical Considerations.
Te animal research guideline adopted by the ethics committee of the Research Unit in Applied Microbiology and Pharmacology of Natural Substances-University of Abomey-Calavi was followed to ensure adherence to experimental guidelines and animal welfare.

Statistical Analysis.
Te variance was analyzed using the procedure of generalized linear model with the R software version 4.2.0. Te signifcance (p < 0.05) of the group factor was determined using the F test, and the averages were compared two-by-two using the student test.
Te favonoid contents in aqueous extract were higher than favonoid of ethanolic extract ( Table 2).

Antioxidant Activity.
Free radical scavenging depends on the concentrations of gallic acid, butylated hydroxytoluene, and quercetin. Tese curves allowed the determination of the concentrations of each synthetic compound which allows the scavenging of 50% (IC 50 ) of DPPH free radicals. It turns out that the concentrations allowed to trap 50% of the DPPH radicals by the reference compounds are, respectively, 10.45 ± 1.59 μg/mL; 29.98 ± 1.91 μg/mL; and 71.67 ± 2.52 μg/mL for quercetin, gallic acid, and butylhydroxytoluene. Ten, it makes sense that quercetin had higher activity than gallic acid and BHT (Table 3).

Acute Toxicity.
Te mean values of several hematological parameters, namely, white blood cell (WBC) number, red blood cell (RBC) count, hemoglobin (Hb), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), packed cell volume (PCV) did not (p > 0.05) diverge signifcantly in all three groups. MCV of group 2 varied signifcantly at day 14. Similarly, the thrombocytes were signifcantly increased on day 14 in all three groups, especially in the two experimental groups (Table 4).
We found a signifcant diference (p ≤ 0.05) in ALT levels and a very signifcant diference (p ≤ 0.001) in alkaline phosphatase (ALP) levels. However, AST and ALT levels at the 14 th day for all groups were slightly lower than the ones at the beginning. At the same time, creatinine and urea levels showed no signifcant (p > 0.05) change from day 0 to day 14. However, we noted a slight increase in the creatinine concentration at day 14 in group 1 and a slight decrease in groups 2 and 3; whereas, urea levels showed a decrease at day 14 in groups 1 and 3 and an increase in group 2 (Table 5). Figure 1 shows the weight variation between the experimental rats and the controls. At the beginning of the experience, the average weight in the three groups did not vary signifcantly. However, we found a signifcant increase in the average weights during the test. Te rats that received aqueous extract showed the highest weight gain (198 g). Te control rats showed the lowest weight gain (175 g). Figures 2 and 3 show the results of histological sections of the kidney and of the liver, respectively.

Histological Sections.
Te diameter of the centrilobular vein was enlarged in rats having received the S. italica aqueous extract, whereas in rats treated with the ethanolic extract, this vein was narrowed. In all experimental rats, the volume of hepatocytes increased (Figure 2).
Analysis of the Figure 3 shows that the renal cortex was normal in all rats. Te glomeruli are normal (compact) in the controls but are altered (red arrow) with the retrieval of some podocytes and increase of Bowman's space in the batches that received the extracts. Te renal tubules (black arrows) in the controls had a normal morphology. Teir volume had increased in the experimental rats. Te tubular epithelial cells (yellow arrow) were detached from the basal lamina and several of them were necrotic (Figure 3).

Discussion
Te main organ managing vital functions like digestion and detoxifcation of molecular compounds in the body is the liver [1]. In developing countries, the use of pharmacopeia therapies is very common because of the fora wealth. However, the chemical content and safety of certain plants used remain unknown. Tis work has looked for the chemical composition, the antioxidant power, and the acute oral toxicity of S. italica leaves extracts. Phytochemical analysis of S. italica leaves powder showed steroids, alkaloids, tannins, favonoids, saponosides, mucilages, quinone derivatives, and cardiotonic derivatives. On the other side, we did not fnd any traces of free anthracenes and cyanogenic derivatives. Tese results support the ones of [19] who also detected favonoids, alkaloids, and steroids presence in the aqueous and methanolic extracts of S. italica. Tannins were not found in Dabai et al. [19], as opposed to our study. Tis could be due to the variations observed in the nature of the plant material, phenological state, and geographical location [19]. Te present work is not fully in line with the work of Alqethami and Aldhebiani [20] who did not detect favonoids presence in S. italica fruit, but did fnd tannins and saponins as in our study [20].
Te nonappearance of certain secondary metabolites in our results could be justifed by the seasonal variations. Tey can afect chemical composition but also biological activity of plants [21]. Te results of this work agree with those of [22] who did not detect the presence of cyanogenic derivatives in the leaves of S. italica.
Phenolic substances are recognized for their numerous health benefts. Various pharmacological activities (antioxidant, anticancer, and antimicrobial activities) are    [24,25]. Te present work shows a progressive rise in the trapped DPPH percentage. It goes up to 80%. Te aqueous extract of this plant was the most active with an IC50 equal to 10 μg/ mL and ensued by the semiethanolic extract (66 μg/mL) and by the ethanolic extract with an IC50 value equal to 78 μg/ mL. Te concentrations in semiethanolic and ethanolic extracts of S. italica leaves diverged from 9.2 μg/mL to 78 μg/ mL. Tis is not in line with the results of [26] which pointed the ethanolic extract as the best source of antioxidants. Te highest IC50 value was given by the semiethanolic extract. Tis proves the low antiradical activity of semiethanolic extract compared to others. Te aqueous extract showed the highest activity with an IC50 value � 9.2 μg/mL tailed by the ethanolic extract. Te ethanolic and aqueous extracts of S. italica leaves showed higher antiradical activities than the controls used in this work.
Flavonoids, which pertain to phenolic compounds, are considered as potential antioxidant sources. Tey have the ability to reduce free radical species and reactive forms of oxygen. Te reducing power of free radicals is explained by the shield efect of favonoids. Te low redox potential that makes favonoids thermodynamic has also been attributed to their shielding efect. Te transfer of the hydrogen atom generated by the antioxidant reaction gives rise to a peroxyl radical. According to the work of Mokgotho et al. [27], the antioxidant power of S. italica is linked to its content in resveratrol. For other authors, the use of S. italica in traditional care was due to its antibacterial, antioxidant, antidiabetic, and hypertensive properties [28][29][30][31].
For the acute oral toxicity of S. italica leaves extracts, after gavage of the rats with the ethanolic and aqueous extracts of Senna italica, we found neither mortality nor morbidity signs. Not a single moribund animal was obtained throughout the 14 days of experiment. Tis supports the results of [6]. We did not observe any change in the functioning of the skin, eye, hair, and respiratory system. Behavioral and physical signs of toxicity such as sleep disturbance, seizure, breathing, restlessness, or hyperactivity were also absent. Tese results give evidence for the nontoxicity of the ethanolic and aqueous extracts of S. italica after administration at 5000 mg/kg of body weight. Te oral ingestion of ethanolic and aqueous extracts of S. italica at 5000 mg/kg did not afect the normal growth of the experimental rats as shown by the evolution of the weight gain ( Figure 1). However, a gradual change in their weight was underlined. According to mean weight values analysis, the weights of the rats of the three groups at day 0 and day 14 did not difer signifcantly. Considering the extract impact on weight gain, our results are similar to the ones of Frimpong and Nlooto [29].
To assess the extracts efects on the function of the rats's vital organs, certain hematological parameters were measured. Te white blood cells (WBCs) number, hemoglobin (Hb), red blood cells (RBCs), hematocrit (Hte), the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin content (HCM), platelets (PLTs) were checked for the rats. Table 3 sums up the results obtained about the hematological parameters. We found that the count of white blood cells (WBCs), hemoglobin (Hb), red blood cells (RBCs), hematocrit (Hte), and mean corpuscular hemoglobin concentration (MCHC) did not increase signifcantly (p > 0.05) in all three groups as opposed to mean corpuscular volume (MCV) which increased on day 14 compared to day 0. Tere is a decline in the count of red blood cells (RBCs), an anemia and an increase in case of exaggerated production or loss of liquid [32]. Platelet count (PLT) increased signifcantly on day 14 compared to day 1 in group 1 (p ≤ 0.05). It also increased very signifcantly (p ≤ 0.0001) in the two other groups on day 14. Te platelet count allows the detection of a bleeding, infectious, or infammatory risk after huge bleeding [32]. Te thrombocytes count variations observed in our case would be linked to physiological growth of the rats. Statistical analysis showed no signifcant diference (p > 0.05) between the hematological parameters of experimental rats having received the aqueous extract or the ethanolic extract and control rats for the other parameters. Tere was also no signifcant variation in these parameters in each group between the frst day and the fourteenth, despite the frst observations. Te high values of MCV, CHM, and MCHC indicate the presence of macrocytic normochromic red blood cells, while a decrease points to the presence of hypochromic microcytic RBC [30]. Apart from the signifcant variation in MCVs observed in batch 2 on day 14, in our case, there was no other signifcant behavior. Tis shows that the RBC of the rats were normocyticnormochromic and, therefore, that the ethanolic extract had no noxious efect on the RBC of the rats at 5000 mg/kg of body weight. In conclusion, the ethanolic and aqueous extracts of S. italica had no toxic efect on blood platelets and the aqueous extract of S. italica had no toxic efect on MCV. However, various factors related to the subject and its environment could be responsible for the nonsignifcant variability recorded in this study and the observations related to the cell variability, in particular lifespan [33].
Several biochemical markers, namely, glucose, transaminases (AST and ALT), urea, creatinine, alkaline phosphatase (ALP), and gamma GT were also measured to assess the impact of ethanolic and aqueous extracts of S. italica on rats' vital organs. Table 5 shows the average concentrations of these parameters in the experimental group with the time. We obtained no signifcant diference (p > 0.05) between the levels of these markers in the groups on day 0 and 14, except on day 14 for group 2 (Table 5). Tere was a substantial variation (p ≤ 0.05) in ALT level and a highly signifcant diference (p ≤ 0.001) in alkaline phosphatase (ALP). However, AST and ALT levels on day 14 for all groups of rats were slightly lower than on day 0, as were creatinine and urea levels. Te fndings revealed no signifcant change (p > 0.05) between day 0 and 14. However, we noted a slight increase by day 14 in the creatinine level in group 1 and a slight decrease in groups 2 and 3, while urea levels decreased in groups 1 and 3 and increased in group 2. Creatinine and urea levels are good indicators of kidney function [34].
Te data collected about biochemical parameters revealed no signifcant variation between the levels of AST, blood glucose, urea, gamma GT, and creatinine for the experimental rats exposed to the extracts compared to controls. Among the biochemical markers covered, transaminases (AST and ALT) are normally found in many cells' cytoplasm and mitochondria, mainly in the liver, heart muscle, and skeletal muscle. However, their concentrations are lower in the pancreas, kidney, and erythrocytes [35]. Terefore, increased serum AST and ALT levels indicate liver toxicity. Tis occur generally in the blood when the permeability of liver cells is impaired or when necrosis take place.
AST and ALT are hepatic enzymes insuring the chemical transfer of an amine group to other molecules in the liver [34]. Te activity of these enzymes is relative to the degree of damage [35]. Terefore, they are two relevant indicators of hepatotoxicity [36,37].
An increase in two-or three-folds range of ALT levels implies hepatic cytolysis [32]. An increase in serum AST activity indicates a traumatic, an infammatory or degeneracy due to plasma membrane damage and cellular necrosis [37,38]. Te signifcant change in ALT levels seen in group 2 highlights a hepatic cytolysis. Infammation and tissue degeneration noticed in rats treated with S. italica aqueous extract at 5000 mg/kg of body weight did not afect hepatocytes. Terefore, we can deduce that the S. italica ethanolic extract did not cause any damage on the liver of rats at 5000 mg per kg of body weight.
Renal function was assessed through serum ureal and creatinine concentrations. Creatinine and urea are important markers of the kidney function [39,40]. Tese metabolism products have a constant level under normal conditions [41]. Renal impairment is refected by their decrease or increase [42]. Pritchard and his colleagues had shown that a reduction in serum creatinine could indicate cachexia. Looking at the serum urea concentration, its rise can sign a nephropathy, dehydration, electrolyte imbalance, Journal of Toxicology 7 hypo-albuminemia, and tissue catabolism [42]. Because these parameters did not vary signifcantly in experimental rats as opposed to controls, we came to the conclusion of a normal kidney function. In sum, since no signifcant variation in AST, gamma GT, blood glucose, urea, and creatinine levels was noted, we can deduce that the ethanolic extract of S. italica is not toxic to the liver and kidneys in rats at 5000 mg/kg of body weight. S. italica ethanolic extract administered to rats in a proportion of 5000 mg/kg of body weight have not caused concomitant changes in the number of white blood cells and red blood cells. Te frst are essential to fght infection and develop resistance to infection after a prior exposure or vaccination. Tey consist of monocytes, lymphocytes, and granulocytes [43]. In the case of infection, infammation, cancer or leukemia, the leucocytes count is increased and can be reduced by the bone marrow failure, liver disease, splenomegaly, autoimmune diseases, and certain drugs. Teir levels are not very diferent in treated rats in comparison to the control group. Tis demonstrates that the S. italica ethanolic extract presents no toxicity for the white blood cells of rats at 5000 mg/kg of body weight. S. italica aqueous extract administered at 5000 mg/kg body weight produced signifcant change in ALT levels indicating liver cytolysis, infammation, tissue degeneration, and thrombocytosis in administered rats. Tis explains the switch of the hemostasis system. Moreover, there was a signifcant variation in the alkaline phosphatase level of the rats having received 5000 mg/kg of body weight of S. italica aqueous extract. We can then summarize that the ethanolic extract was not toxic at a limit dose of 5000 mg/kg of body weight, while the aqueous extract of S. italica would be toxic only at a dose lower than 5000 mg/kg of body weight.

Conclusions
Te Senna italica leaves contain various pharmacologically active compounds including phenolics, derivated quinone, and favonoids, which are good antioxidant, anti-infammatory, and protective. S. italica leaves are nontoxic at the dose of 5000 mg/kg of body weight. Tus, the leaves of S. italica have a powerful antioxidant activity. Tey can be used orally as traditional medicine. S. italica leaves constituted then a useful phytobiotic resource which can promote human health. Dose studies are, however, necessary depending on the type of pathology.

Data Availability
No additional information is available for this paper.

Conflicts of Interest
Te authors declare that they have no conficts of interest.

Supplementary Materials
EQUATOR network study checklist. (Supplementary Materials)