Evaluation of the Biological Efficacy of the Aqueous Extract of Cyperus Rotundus on the Toxic Effect of Sodium Benzoate in Albino Male Rats

EurAsian Journal of BioSciences Eurasia J Biosci 14, 7481-7486 (2020)

Evaluation of the biological efficacy of the aqueous extract of Cyperus rotundus on the toxic effect of sodium benzoate in albino male rats

Ruqayah Abbas Jaddoa 1, Naseer Marza Hamza 1* 1 Department of biology, College of Education for Pure Sciences, University of Kerbala, IRAQ *Corresponding author: [email protected]

Abstract The current study showed that dosing animals with sodium benzoate at a concentration (200mg / kg / bw) for 30 days caused tissue damage to the liver, which was the occurrence of congestion and expansion in the central vein, necrosis of hepatocytes and their degeneration with severe irregularity in the arrangement of the hepatic cords, as well as the presence of Congestion and expansion of the sinuses with congestion of the central vein and infiltration of inflammatory cells around it, while treatment with aqueous extract of the leaves of the Cyperus rotundus in three concentrations (250- 300-350) mg / kg of body weight reduced or suppressed the toxic effects of benzoates. The lack of expansion of the sinuses and the return of most hepatic cord cells to regularity with the presence of degeneration in some hepatocytes in the first treatment group, but in the second and third focus groups, the central vein congestion disappeared in a way and the expansion of the sinuses decreased with the regularity of the hepatic cords and the tissue became as close as possible to the normal tissue. Noting that there is slight degeneration in some cells and thus it can be considered that the aqueous extract of the Cyperus rotundus has biological efficacy in resisting the histological changes of the liver resulting from Take poisons intentionally or unintentionally, with the possibility of being considered safe for use.

Keywords: Sodium benzoate, Cyperus rotundus, pathological changes

Jaddoa RA, Hamza NM (2020) Evaluation of the biological efficacy of the aqueous extract of Cyperus rotundus on the toxic effect of sodium benzoate in albino male rats. Eurasia J Biosci 14: 7481-7486.

INTRODUCTION abundant flowers with few seeds and spikes are lanceolate or oval. Arranged radially, Cyperus rotundus Cyperus rotundus Plant L is a species known for its medicinal benefits within this The plant Cyperus rotundus is one of the plants that genus (Chakravarty, 1976). has been used since ancient times in the treatment of This plant was given many generic names, and many diseases for its medicinal benefit. Its tubers have among its most popular common names in English is been used in Egypt since the ancient Stone Age in the (Nut-grass) which is the most used as it was called treatment of various diseases, including as an anti- (Purple Nut-Sedge, Muthan, Tirirca and Musta). Arabic bacterial, pain reliever, fever reducer, gas repellent, is (Cyperus rotundus), (Al-Sajil), (Swan) and (Sale), and diuretic, immune stimulator, regulator For the menstrual the most common of them is the Cyperus rotundus cycle, reducing body weight, killing cancer cells (El- (Bajpay et al., 2018). Moghazy, 1967). Researchers (Tyler, Brady and Robbers, 1988) This plant possesses a great capacity to grow in indicated that the Cyperus rotundus plant contains many various forms of environmental conditions as it invades effective compounds that play a fundamental role in lands in mountainous regions in northern Iraq as it grows treating many diseases, including (polyphenols, in plateaus and plains and in marshes, swamps and flavonoids, essential oils, ascorbic acid, linoleic acid, central regions. It also grows in the sedimentary plains Pectin) Thus, this plant is considered a pain reliever, and on the banks of rivers (Al-Ali, 1977). anti-bacterial, anti-malarial, sugar-lowering, immune The genus Cyperus L. is one of the most important stimulant, and hair tonic. genera of the family Cyperus rotundusia and the origin of this name Cyperus is Greek. This genus includes Received: February 2020 many species in which the plant consists of tubers that Accepted: April 2020 grow under the soil and a triple stem in cross section and Printed: December 2020

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Researchers (Duke, 1992) also indicated that the the growth of these organisms through the interference roots of the Cyperus rotundus plant contain multiple that occurs between chemical preservatives and phenols that contribute to the prevention of cancer and membranes. Microorganism cells and their enzymes can be considered as antioxidants, in addition to the and their genetic mechanisms (Sunitha and Sagar, tubers containing essential oils, including Limonene, 2012) . which have medicinal efficacy against breast and Sodium benzoate is one of the dietary additives pancreatic and prostate cancer diseases. because it is has a high inhibitory effect on the growth of Many researches indicated that the extract of the many yeasts and molds, in addition to a wide range of Cyperus rotundus plant contains small quantities of bacteria, but in addition to its inhibitory effectiveness, essential oils, which constitute (0.3 - 1%) of its crude many studies have indicated that it has harmful effects weight, and these oils contain at least (27) medically on the liver, the researchers pointed out ,In their study effective compounds, including derivatives, of the effect of oral dosing in adult male rats with sodium Sesquiterpene epoxides, Sesquiterpene hydrocarbon, benzoate, it has clear effects on liver tissue, and the monoterpenes, aliphatic alcohols, and sitosterol (Dhillon level of these effects is significantly correlated with the et al., 1993). length of treatment, so pathological changes were Researchers (Bajpay et al., 2018) indication in their observed in the liver represented by the loss of most of study of the medicinal efficacy of the aqueous extract of the structural organization of the tissue with the Cyperus rotundus L, that there are widespread presence of ulcers in the hepatocytes and the applications of this plant in the medical and cytoplasm. In addition to the death of some cells and pharmaceutical fields as it is used in the treatment of their decomposition with fatty changes in the diseases of indigestion, constipation, flatulence and pain hepatocytes represented by the presence of fatty Stomach, as it is anti-microbial and anti-inflammatory droplets in the cytoplasm as well as the occurrence of and contributes to increasing wound healing activity and depletion of carbohydrates, this is in the experiment resistance to cancer and oxidative stress. groups for the first time period, As for the changes that The researchers (Sivapalan and Jeyadevan, 2012) occurred after the end of the second period of time, the indicated that the Cyperus rotundus plant contains many liver cells were completely destroyed and the normal effective compounds that contribute to the treatment of shape of the liver lobules was lost (Khidr, Makhlouf and many diseases, including diseases resulting from the Ahmed, 2012). occurrence of blood infections, which are accompanied In the same direction, the study (Sinha and D’Souza, by the occurrence of the release of free radicals, which 2010) indicated that the oral treatment of white mice with constitute the stress of oxidative stress, which in turn is sodium benzoate resulted in severe hepatocyte related to infection with many Diseases that result in vacuolization, loss of nucleation in some sites, increased damage to a large group of body tissues, which comes mitochondrial activity, plasma membrane shrinkage and in the forefront of liver tissue, where all the toxic effects the formation of finger-shaped extensions. that the body is exposed to, intentionally or Aim of the study unintentionally, are treated. The present study aims to demonstrate the In the same direction, researchers (Soltan and Zaki, protective effects of Cyperus rotundus L aqueous extract 2009) indicated that the medicinal efficacy of Cyperus against the toxic effects of sodium benzoate on liver rotundus L comes from the ability of this plant to resist tissue. oxidative stress through the formation of free radicals inside the body. The alcoholic extract of this plant was MATERIALS AND METHODS tested on cells of type (neuro-2a). It was found that it has antioxidant activity as well as anti-tumor activity in the The current study was conducted in the animal body. house of the Department of Life Sciences / College of Education for Pure Sciences / University of Karbala for Food additives the period from November 2019 to April 2020, in which Additives are defined as substances that are added male Norwegian white rats used the type Rattus to food and become one of its components, directly or norvegicus. The animals were deposited in the animal indirectly, and affect its properties, and include any home to adapt to laboratory conditions before their use. substance used in the production, manufacture, They were placed in plastic cages measuring (20 x 40 x processing, packaging, transport and storage of food 20) cm for the length, width and height respectively, and (James, 2004) . each cage had a metal cap with a clamp provided with a The reason for the use of preservatives is due to the water bottle and a place to put food, and the floor of the presence of microorganisms that cause food spoilage cages was spread with sawdust that was replaced every and other materials and to cause unwanted changes in two days to keep the rats clean while sterilizing them the materials, as their use leads to delay or prevent the from time to time With sterilizers, the animals were freely occurrence of unwanted changes in food, by inhibiting

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slides were prepared from them, based on the method of (Bancroft and Stevens, 1982).

RESULTS First: The effect of treatment with sodium benzoate on the histological structure of the liver compared to the control group: The liver generally consists of two main lobes, a right lobe, which is the largest and a left lobe, and each lobe in turn is divided into smaller lobes called lobules, where the cross sections of the liver tissue in the control group showed that each lobe contains a central vein surrounded by a group Large, compact hepatic cells that are arranged in strips and contain clear nuclei, between supplied with water and ad libitum during the which are gaps called sinusoids (Figs. 1 and 2) experiment. Whereas liver tissue sections treated with sodium Design of the experiment benzoate at a concentration of (200) mg / kg for a period The experiment included a study of 30 rats an of 30 days showed that he suffers from congestion and inventory whose ages ranged between (25-27) weeks expansion in the central vein, necrosis of hepatocytes and average weights (300 - 400 grams). The animals and their degeneration with severe irregularity in the were divided into five groups, with six animals for each hepatic cords as well as the presence of congestion and group. The first group was considered a negative control expansion of the sinuses. Central and infiltration of group and dosed orally with tap water only. And the inflammatory cells around it (Figs. 3-5). second group was a positive control group and was Second: The protective efficacy of the aqueous dosed orally with sodium benzoate at a dose of (200 mg extract of the Cyperus rotundus plant compared with / kg / bw) and the third, fourth and fifth groups were the positive control group: considered protective groups and a dose of all with The results of the current study of the cross-sections sodium benzoate in the same previous dose in addition of the liver tissue of the samples treated with Cyperus to the aqueous extract Leaves of the Cyperus rotundus rotundus at leaf extract as a protective substance and plant, at a dose of(250 ,300,350mg / kg / bw) for the last with concentrations (250-300-350 mg /kg of body three groups in a row and after the end of the experiment weight) showed that they were less affected by the period of (30) days, the animals were anesthetized and toxicity of sodium benzoate when compared to the dissected, the liver extracted and preserved with a samples of the group of rats treated with sodium formalin solution of 10% concentration until the tissue benzoate at a concentration of (200 Mg / kg of body

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the hepatic cords, as well as degeneration of hepatocytes in the prophylactic group, the concentration of (250 mg / kg of body weight), while it was observed in the prophylactic group With a concentration of (300 mg / kg of body weight), it was less congestion to the central vein, expansion of the sinuses, regularity of some hepatic cords, and degeneration of some hepatocytes, while in the prophylactic group at a concentration of (350mg /kg of body weight), it was observed that liver tissue Closer to normal, with reduced central vein congestion, which appears naturally with the presence of sinuses and regularity of the hepatic cords. It is noticed normal in most hepatic areas and slight degeneration of other areas of the liver (Figs. 6-8).

DISCUSSION

The results of the current study showed that there is a clear significant effect on liver tissue in the group treated with sodium benzoate, and that is consistent with the researchers (Kaboglu and Aktac, 2002) indicated in their study, which showed degenerative and necrotic changes in liver tissue in animals Treatment with sodium benzoate, which was detected by a close physical examination. In the same direction, the researchers (13) indicated in their study of rat liver tissue that was treated with sodium benzoate at a concentration of (1) g / kg for a period of 3 months. Normal and completely broken liver cells in some liver lobules. The researchers (Smyth Jr and Carpenter, 1948) weight) was observed less congestion of the central vein indicated in their study of the toxic effects resulting from and expansion of the sinuses with irregularity of some of treatment with sodium benzoate at concentrations of

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and then the decrease in the weight of the organs (liver, kidneys, muscle mass), which in turn is associated with a decrease in body weight. On the other hand, the results of the current study proved that the aqueous extract of the leaves of the Cyperus rotundus plant had a protective effect against the toxic effects that the body was exposed to, as it contributed significantly to treating most of the tissue damage that the body was exposed to as a result of exposure to sodium benzoate and that this resulted from the containment of this extract on Many active substances with medicinal efficacy (polyphenols, flavonoids, essential oils, ascorbic acid, linoleic acid, pectin) (Chandratre, Chandarana and Mengi, 2012). In the same direction, the researcher (Necib, Bahi and Zerizer, 2013) pointed out that the significant preventive effect of the Cyperus rotundus plant extract contributed to improving the level of the tissue structure of the liver in the extract treated with the extract and that the level of protection was significantly associated with the increase in the concentration of the extract with certain limits and that this role results from the ability of

this plant To improve the level of enzymatic and non- 640 and 1320 mg / kg for a period of 90 days in the enzymatic antioxidants due to what this plant contains of inventory of a mortality rate equivalent to 50% effective compounds and thus reduce the level of accompanied by the occurrence of changes and oxidative stress that the body is exposed to by curbing pathological signs in the tissue of the kidney and liver the formation of effective oxygen cells that are when examined Histologically. generated inside the body from the ingestion of toxic In their study of the effect of dietary additives, the substances directly or indirectly. researchers (Aldaamy and Al-Zubiady, no date) The researchers (Bahi and Necib, 2014) in their indicated that dosing rats with these substances for a study of the medicinal effects of the Cyperus rotundus long period of time led to the occurrence of pathological plant, indicated that there was a clear change in the changes in the blood followed by histological changes tissue-damaged liver cells when the extract was taken that occur mainly in the liver tissue, which accompanies through a decrease in the level of venous hemorrhage the change in the oxidation parameters inside the body and the accumulation of inflammatory cells with where the metabolism of toxic substances leads to The regularity of the hepatic cords after they were exposed release of high concentrations of the active oxygen to great damage as a result of treatment with mercury species (ROS) as part of the metabolism process, and chloride. Which caused various pathological changes in as a result of the formation of ROS. SOD and GSH are the livers of the treated rats. consumed in order to prevent cell death by the effect of Researchers, (Kamala et al., 2018) In their study, free radicals. Which leads to damage to the tissues of the major role of the Cyperus rotundus plant root extract the liver and kidneys. as an antioxidant comes from its high content of In the same direction, (Azez and Hamza, 2020) flavonoids, ascorbic acid, and other active ingredients. indicated in their study of the effect of dietary additives, The researchers also indicated (8) that Cyperus which were exemplified by sodium fluoride, that rotundus L extract is widely used in a range of medicinal exposing the body to these substances in high and and pharmaceutical applications, which are widely used unexamined concentrations by eating lunch directly or in traditional medicine around the world to treat various indirectly would cause a clear decrease in body weight, diseases and ailments such as indigestion, constipation, which is mainly related to tissue damage. Which comes flatulence. Nervous stomach pain, skin diseases, boil in the forefront of the liver and kidneys, as the liver and infections, staph infections and stomach pain. It also has kidneys take it upon themselves to rid the body of all a variety of pharmacological properties such as toxic elements, which exposes them to harm, whether antibacterial and microbial activity, has anti-platelet on the part of calibrators, physiological or activity, anti-inflammatory activity and has anti- histopathology, which resulted in the lack of drinking rheumatoid, anti-diabetic, anti-cancer pharmacological water in experimental animals, and thus an imbalance in properties. Anti-malarial, anti-oxidant, anti-obesity, anti- bodily fluids and a lower rate The metabolism of animals, liver damage.

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In addition, the researchers (Sivapalan and stress from the metabolism of toxic substances inside Jeyadevan, 2012) in their study of the protective role of the body. the alcoholic extract of Cyperus rotundus plant indicated that the extract showed great effectiveness in reducing CONCLUSION liver enzymes levels after they were significantly the aqueous extract of the Cyperus rotundus has elevated in mice treated with sodium fluoride and that biological efficacy in resisting the histological changes of this reduction in these enzymes was followed regularly. the liver resulting from Take poisons intentionally or The histological composition of the liver lobes after they unintentionally, with the possibility of being considered were exposed to tissue damage as a result of oxidative safe for use.

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