The therapeutic potential of
There is currently much interest in the therapeutic potential of traditional and herbal therapy as an antioxidants in reducing free radical-induced oxidative stress and inflammation particularly in hepatic diseases [
This causes the generation of ROS such as hydrogen peroxide (H2O2) and hydroxyl (OH−) radicals that affect the cellular membrane and induce lipid peroxidation by eliminating hydrogen from a polyunsaturated fatty acid and subsequent liver damage or necrosis [
Medicinal plants are an important source of natural antioxidant agents because of the less toxic nature and being free from side effects compared to synthetic antioxidant [
The matured pods of
The powdered seeds (150 g) were soaked in methanol (500 mL) for 7 days under room temperature, 28°C [
The total phenolic content of the extracts was determined using the method described by Li et al. [
The DPPH radical scavenging activity of
The percentage DPPH radical scavenging was calculated using the following equation:
The assay was conducted based on the modification method by Chakraborthy [
The reducing power assay was evaluated by the method of Oyaizu (1986) as described by Yildirim et al. [
The IC50 is defined as the concentration of the sample that provides inhibition of 50% of the initial radical concentration with the unit, mg/mL or
Fifteen adult male Swiss albino mice aged 6 to 8 weeks old and weighed 25 to 30 g were used to study the hepatoprotective activity of
The paracetamol tablets were obtained from a nearby pharmacy. Each tablet contains 500 mg of paracetamol. The mice were administered with paracetamol at a dose of 1 g/kg body weight (b.w.). The paracetamol was made into fine powder using a mortar and pestle. The powdered paracetamol was suspended in distilled water and was administered according to the body weight of mice. An aqueous suspension of seed extract was prepared in distilled water and different doses of
Fifteen adult male Swiss albino mice were divided into 5 groups and each group consists of 3 mice each (Table
Mice groupings and administrated treatments.
Groups | Treatment |
---|---|
Negative control | 10% DMSO |
Induced | 1.0 g/kg paracetamol per body weight |
Treatment Group I | 250 mg/kg seed extract per body weight + paracetamol |
Treatment Group II | 500 mg/kg seed extract per body weigh + paracetamol |
Positive control | 200 mg/kg silymarin per body weight + paracetamol |
15 adult mice were divided into 5 groups (
The mice of each group were anaesthetized with ketamine/xylazine and blood was collected directly from the heart. Then centrifuged at 3000 rpm for 15 minutes to separate the serum and kept at 4°C for analysis of various biochemical parameters including ALT, AST, and ALP [
The mice were weighed daily during the study and the body weights of the mice were determined and recorded. After the mice were euthanized, the livers were isolated and washed with saline and weights were determined by using an electronic balance [
Livers were perfused with saline and homogenized in chilled potassium chloride (1.17%) using a homogenizer to determine the in vivo antioxidant level in the liver tissues.
The liver samples of the mice were fixed in 10% buffered formalin. After fixation, the livers were dehydrated in a graded series of alcohol, cleared in xylene, and embedded in paraffin wax. Multiple 5
Data are expressed as mean ± Standard Deviation (SD). Significance was evaluated using
The extraction was carried out using matured
The total phenolic content of the
Figures
Percentage inhibition of methanolic seed extract of
Inhibition effect of
Dose-dependent NO scavenging activity of the methanol extract of the
Percentage inhibition of methanolic seed extract of
Inhibition effect of
The dose-response action for the reducing activity of methanol extract of
Reducing power of methanolic seed extract of
Table
Effect of
Groups | Dose (mg/kg) | Body weight, BW (g) | Liver weight, LW (g) | Relative liver weight (%) |
---|---|---|---|---|
10% DMSO pretreated negative control | — | | | |
Paracetamol-induced | — | | | |
Silymarin + paracetamol | 200.00 | | | |
Seed extract + paracetamol | 250.00 | | | |
Seed extract + paracetamol | 500.00 | | | |
Results are expressed in means ± SD (
The effect of
Effect of
Groups | Dose (mg/kg) | ALT (U/L) | AST (U/L) | ALP (U/L) |
---|---|---|---|---|
10% DMSO pretreated negative control | — | | | |
Paracetamol-induced group | — | | | |
Silymarin + paracetamol | 200.00 | | | |
Seed extract + paracetamol | 250.00 | | | |
Seed extract + paracetamol | 500.00 | | | |
Results are expressed in means ± SD (
The in vivo antioxidant level in the liver tissues of paracetamol-intoxicated mice pretreated with
Effect of
Groups | Dose (mg/kg) | GSH (mg/mg protein) | SOD (U/mg protein) | MDA (nmol/mg protein) |
---|---|---|---|---|
10% DMSO pretreated negative control | — | | | |
Paracetamol-induced | — | | | |
Silymarin + paracetamol | 200.00 | | | |
Seed extract + paracetamol | 250.00 | | | |
Seed extract + paracetamol | 500.00 | | | |
Results are expressed in means ± SD (
To further explore the biochemical analysis findings, histopathological observation was conducted on liver tissue. Liver sections taken from paracetamol-induced mice (Figure
Light microphotograph of negative control liver. (H: hepatocytes; N: nucleus; SS: sinusoid; CV: central vein). Magnification: 40x.
Light microphotograph of paracetamol-induced liver. (NP: neutrophil; DS: dilated sinusoid; FN: focal necrosis; CV: central vein). Magnification: 40x.
The hepatocyte nucleases are at a recovery stage and there are very minimal numbers of neutrophils surrounding the central vein.
The pretreated group with seed extract at a dose of 500 mg/kg b.w. (Figure
Light microphotograph of liver cells of mice treated with
Light microphotograph of liver cells of mice treated with
Light microphotograph of liver cells of mice treated with silymarin (H: hepatocytes; N: nucleus; SS: sinusoid; CV: central vein). Magnification: 40x.
This study showed that
The result of the present study clearly indicated that the methanol extract of
The DPPH radical is regularly used as a substrate to evaluate the antioxidant activity of medicinal plant; it is a stable free radical that can receive an electron to become a stable molecule. The
Nitric oxide (NO) is a key chemical facilitator involved in the regulation of various physiological processes. Higher concentration of NO is related with numerous diseases. Oxygen molecules react with the excess amount of NO to generate nitrite and peroxynitrite anions, which eventually act as free radicals [
Reducing power assay is commonly used to assess the capability of a natural antioxidant to donate an electron [
The body weight of the mice in this study was not affected by the treatment of paracetamol, silymarin, and
Paracetamol induced mice liver damage in this study because of the formation of toxic metabolite, NAPQI, which is initially detoxified by conjugation with glutathione to form mercapturic acid [
However, pretreatment of high dose of
The pretreatment with the 250 mg/kg b.w. dose of seed extract slightly reduced the elevated serum liver enzyme levels but not as effective as the treatment with 500 mg/kg b.w. dose of the seed extract. This proved that
This study also showed that in mice group with paracetamol-induced liver hepatotoxicity, the ALT and AST levels were elevated more compared to the lesser increase in the ALP level. This finding indicated that paracetamol caused cell necrosis at a single dose of 1 g/kg b.w. as cell necrosis involves the initial increase of ALT and AST and modest increase in ALP. The ratio of ALT/AST : ALP plays an important role in the determination of the types of liver damage by hepatotoxins. The ratio is greater than or equal to five for cell necrosis injury compared to if the ratio is less than or equal to two for cholestasis injury.
The ROS induced oxidative stress inside the cells triggered damage to hepatic parenchymal cells leading to hepatic injury [
The methanolic extract of
The seed extract which exhibited a concentration dependent activity was not effective in recovering histopathological lesions at a dose of 250 mg/kg b.w. as
In the present study, the
The methanolic extract of
In conclusion, the results of the present study strongly suggested that
The authors have no conflict of interests to declare concerning this article.