@2024 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2016;22(3):185-191

Background
The liver is one of the vital organs of human body that detoxifies external substances, drugs, etc. Meanwhile, it may be damaged and liver diseases would appear [1].
Previous Studies
... [2]. Liver damage is usually associated with cell necrosis, high tissue peroxidation and reduction in the level of tissue glutathione. In addition, serum levels of biochemical indices such as SGPT (serum glutamic pyruvate transaminases), SGOT (serum glutamic oxaloacetic transaminase), ALP (alkaline phosphatase), GGT (gamma glutamyl trans-peptidase) and bilirubin increase [3]. ... [4] In recent years, there has been a tendency toward the use of herbal medicines for treating various diseases [5]. Salvia (Salvia officinalis) has several active compounds such as Toine, Cineol, Borneol, Pinene, Flavonoid, Saponin, Glycosides, resin, vitamin C, vitamin E, tannins, resinous substances and Ditropan [6].... [7-11].
Aim(s)
This study aimed to investigate the therapeutic effects of salvia (Salvia officinalis) on toxicity resulted by acetaminophen poisoning in the liver.
Research type
This study is carried out experimentally in the laboratory.
Research society, place and time
This study was designed and conducted in Toxicology Research Center in Jondi Shapoor University of Ahwaz in spring, 2015.
Sampling method and number
60 albino mice in the weight range of 20 to 25 grams were used. They were bought from the laboratory animal breeding center in Jondi Shapoor University of Ahwaz.
Used devices&materials
Here is the way of preparing the extract: first the flowers and aerial parts of the plant were collected at flowering season (spring) from Larestan area in the south of Fars province and after being detected by the Medicinal Plant Research Center of Jondi Shapoor University of Ahwaz they were dried and then ground. Then, the ground plant was put into 80%hydro-alcoholic solvent for 3 days in Toxicological Research Center of Ahwaz University and its extraction was taken. After that, the extract was filtered and the filtered solution was distilled by soxhlet (HiDolph Persia; Germany). Then, it was placed in the oven in 30-40 ° C so the dried extract would be prepared. The output of extraction method was 16%. For being used in the experiments, each time a specific amount of the extract was dried, weighted and dissolved in physiologic saline. Acetaminophen Suspension technique: Acetaminophen powder was prepared in the commercial company (Sigma; USA). Since acetaminophen does not dissolve in cold water, the physiologic serum at 70 °C was used to make a homogenous suspension of the drug. Then it got cooled to 37 °C to be administered [12, 13]. Acetaminophen was prescribed orally by gavage. Studied groups: Animals were divided into 6 groups; each was composed of 10 mice. The first, second and third group each received physiologic saline (negative control), raw extract of salvia (500 mgs per kg) and 500 mgs of acetaminophen per kg in a single dose as a positive control, respectively. The fourth, fifth and sixth groups received doses of 125, 250 and 500 mgs per kg of salvia extract for 5 days, respectively. Then one hour after administrating the last dose of the extract, they received 500 mgs per kg of acetaminophen. The salvia extract was also fed to mice through gavage. Gavage technique: the flexible pipe was attached to an insulin syringe containing salvia extract instead of the needle and it was used for administering drugs to animals. To do this, the skin of the back of animal`s neck and ears was removed so the animal would stay vertically. This led to the open mouth and stillness of the mouse; thus, the gavage tube was put into the mouth such that it could reach the pharynx. After making sure that gavage tube is in the esophagus, the plant extract was given to the animal orally. Sampling: 24 hours after receiving the last dose of acetaminophen, all the groups were anesthetized by chloroform in desiccator. Then blood samples were taken from the mice`s carotid, and their livers were separated for histological studies and were placed in 10% formalin solution. The collected blood was put in a water bath at 37 ° C for 40 minutes to be clotted. Then, for separating the serum, 16-branch locked centrifuge (Behdad; Iran) at 2,500 rounds per minute was used for 10 minutes. The activities of Alanine (ALT) and Aspartate (AST) aminotransferase enzymes were measured through the method by Reitman and Frankel [14]. Alkaline phosphatase enzyme (ALP) was evaluated using the King method [15]. Direct and total bilirubin was measured through the method by Watson and Rogers [16]. The liver tissue was set in formalin solution after being removed and then the tissues were dehydrated using the successive stages of ethanol exposure. Dehydrated liver tissue was fixed in dissolved paraffin, and cuttings with the diameters of 5 micron were made out of it after being molded by rotary microtome device. After fixing the sliced tissues on glass slides, their water yielding and deparaffinization were done using different concentrations of ethanol. Then, the slides were colored by hematoxylin and eosin and finally they were studied using a light microscope (Olympus; Japan). The observed histological changes included necrosis, fatty change, inflammation and accumulation of lymphocytes and the Kuepfer cells. The statistical analysis was performed using SPSS16 software. For each group of the mice, the mean of variable`s level was calculated as statistical mean. To compare the means, one-way Analysis of Variance (ANOVA) was employed, and in order to examine and determine the means` differences and the significance of ANOVA analysis, Tukey complementary test was used in the range of p<0.05.
Findings by Text
After the administration of acetaminophen, the animals suffered from severe liver toxicity which was determined with a significant increase of AST, ALT and ALP enzymes in comparison with the control group (p< 0.05). Moreover, the concentration of direct and total bilirubin went up in this group. Prescribing salvia extract in doses of 125, 250 and 500 mgs per kg reduced AST, ALT and ALP enzymes significantly in groups 4, 5 and 6 compared to the positive control group (p< 0.05). In addition, this reduction was dependent to the doses. Direct and total bilirubin decreased with administrating salvia extract. In the group receiving the extract (500 mg per kg), liver enzymes and bilirubin did not have any significant difference with the control group (Table 1). In histopathological examination of liver tissue, different groups were studied according to the presence or absence of inflammation, the presence of inflammatory cells, fat vacuoles, necrosis and the structure of hepatocytes and liver sinusoids. Histological images of the liver in the control group and the group which had received only salvia extract represent the natural structure of liver cells, hepatic sinusoids and central vein (Figures A1 and B1). However, the group receiving acetaminophen showed severe tissue changes including inflammation, fatty liver, accumulation of lymphocytes and necrosis (Figure C1). Receiving 125 ml grams of the extract reduced the mentioned damages, so that inflammation, necrosis and disorder of lobular were observed in a more limited form (Figure D1). With increasing the dose of the extract a greater improvement was achieved such that at a dose of 500 mgs per kg only inflammation and cell swelling were perceived (Figures E1 and F1).
Main comparison to the similar studies
... [17-30]. In a similar study, Farhoodi et al. investigated the hepatic-protective effect of salvia on the hepatotoxicity resulted by carbon tetrachloride in rats. They administered the extract of the plant to the animals for 14 days and on the fifteenth day, they administered tetrachloride. In this study, indicators such as hepatic transaminases, MDA and superoxide dismutase were used and the groups receiving the extract had a significant reduction in liver enzymes [31] ... [32].
Suggestions
Non-declared
Limitations
Lack of measurement of tissue oxidative stress factors such as MDA, glutathione, catalase and glutathione peroxidase was one of the limitations of this study.
Conclusions
Protective effect of salvia is dose-related, as in a dose of 500 mgs per kg the maximum protective effect can be observed. Although this plant reduces the damage caused by acetaminophen significantly, it cannot completely restore the damage back to normal. Generally, this plant has a potential against the damage caused by acetaminophen in mice.
Acknowledgments
Non-declared
Conflict of interest
Non-declared
Ethical Permissions
This study has been carried out on the basis of the instructions of working with laboratory animals of Ahvaz Medical University.
Funding Sources
This study is financially supported by the Student Investigative Committee of Ahwaz Medical Science University numbered 94s164.

TABLES and CHARTS

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