The present work aims to study the influence of antioxidants activity of lion’s foot ( Alchemilla vulgaris) leaves at different concentrations to give more protection against chronic liver disease. Results indicated that dried lion’s foot leaves had rich in total polyphenolic and flavonoids content (395.65 and 183.10 mg/100g, respectively). These results were reflected to the antioxidant activity (DPPH); it’s noticed that the antioxidant activity of dried lion’s foot leaves was high (131.74%). The major polyphenolic components were benzoic acid (1084.63 ppm) followed by ellagic acid, catechol, and catechin (614.16, 580.54, and 566.53 ppm, respectively) then salicylic acid and protocatechuic acid (479.71 and 444.43 ppm, respectively). On the same trend, flavonoids fractions indicated the highest content in luteo-6-arabinase 8-glucose, apig. 6-rhamnase 8-glucose, acatein, narengin and luteolin (40.01; 15.04; 8.07; 6.64 and 6.42 ppm, respectively). Fifty-six male albino rats were used in biological experiments. Rats fed on basal diet for two weeks before the performance of the experiment. At the beginning, rats divided into eight main group were fed on diets for 45 days as follows: Negative control group (first group) was fed on basal diet. Forty nine rats were fed on basal diet and induced by CCl 4, in paraffin oil (50% v/v, 2 ml/Kg) twice weeks subcutaneous injection to induce chronic damage in the liver, then divided into 7 groups numbered from group 2 to group 8. Positive control group rats fed on basal diet till final experiment (second group). Group 3 and 4 rats treated with 50 and 100 ppm ethanolic leaves extracts, respectively. Also, group 5 and 6 treated with 50 and 100 ppm aqueous leaves extracts, respectively. All extracts were fed on orally every day. While, rats in group 7 treated with 1% and 2% dried lion’s foot leaves. At the end of the experimental period, serums were collected to determine liver and renal functions. The liver was removed surgically for histopathological observation. The results revealed that CCl 4 intoxication impaired liver function. Serum AST, ALT, ALP and total bilirubin levels were elevated by CCl 4 administration, while significant decreasing was noticed in serum albumin in CCl 4 group. Histopathologically, CCl 4 caused congestion of central vain, fatty change of hepatocytes, and focal inflammatory cells in filtration. Treatment with lion’s foot with different forms and concentration attenuated these adverse effects and markedly ameliorated histopathological and biochemical alterations caused by CCl 4 especially with 2% powder and 100 ppm ethanol extract administration. Therefore, the results of this study concluded that lion’s foot can be proposed to protect hepatotoxicity induced by CCl 4 in rats. The results also revealed that the hepatoprotection effect of lion’s foot may be attributed to its antioxidant contents and free radical scavenger effect.
Alchemilla vulgaris (Lion’s foot or Lady’s mantle) is an uncommon herbaceous member of the rose family (Rosaceae). Lady’s mantle is native to Europe and Asia and grows well in New Zealand. The medicinal part of the Alchemilla vulgaris is aerial part with flowers and stems. The recommended official dose of Lion’s foot, the main medicinal plant weigh level is 5 - 10 g/day of the dried plant noticed by [
The liver is a highly sensitive organ which plays a major role in maintenance and performance of the homeostasis in the body. It is the chief organ where important processes like metabolism and detoxification take place. The entry of these toxicants into the body is principally via the gastrointestinal tract and after absorption; they are transported through the hepatic vein to the liver revealed by [
Hepatic disorders are one of the most global health problems, considering drugs, chemicals and/or alcohol as main triggers of the disease [
Alchemilla mollis (Buser) Rothm aerial part and root methanolic-water extracts were evaluated for their hepatoprotective activity on carbon tetrachloride induced hepatotoxicity and hypoglycemic activity on alloxan-induced diabetic mice. Hepatoprotective activity results have revealed that serum ALT levels were significantly lowered by both the aerial part and root extracts at doses of 100 mg/Kg and 200 mg/Kg. Histopathological examination showed that A. mollis aerial parts and roots induced significant recovery from cellular damage; when compared to the carbon tetrachloride group, the most significant activity was observed with A. mollis aerial part extracts at a dose of 200 mg/Kg [
The aim of the present work was to high light the reverse effect of dried Lion’s foot; its aqueous and ethanol extracts at various ratios against the toxicity of CCl4, also to evaluate the various ratios on body weight, feed intake, liver and renal functions and histopathological changes that may occur on rats suffering from chronic liver disease.
Lion’s foot dried leaves was obtained from Horticultural Research Institute, Agriculture Research Center Giza, Egypt. Carbon tetrachloride (CCl4), casein, vitamins, minerals, cellulose and choline chloride will be purchased from El-Gomhoreya Company, Cairo Egypt. Animals: Fifty six male albino rats will be obtained from Helwan breeding farm, Cairo-Egypt. Weighing an average will be between (200 ± 10 g). Casein, cellulose, all vitamins and minerals were obtained from El-Gomhoria Pharmaceutical Company, Cairo, Egypt. Corn oil and starch were obtained from the local market. Kits to determine serum aspartate amine transaminase (AST), alanine amine transaminases (ALT), and alkaline phosphates (ALP), total bilirubin, albumin, glutathione reductase and malonaldehyde were purchased from Biodiagnostic Company in Egypt.
Total polyphenolic compounds were determined by the Folin-Ciocalteau method [
Soluble tannins were evaluated using the Folin Denis method as described by Taira [
Polyphenols and flavonoids compounds determined by HPLC according to method Goupy et al. [
DPPH scavenging activity tests were carried out according to the method of Brand-Williams et al. [
% DPPHradical-scavenging = [ ( AbsorbanceofDPPH − Absorbanceofsample ) / AbsorbanceofDPPH ] × 100
Lion’s foot (Alchemilla vulgaris) leaves were washed in aqueous several times to remove any adhering flesh, dried in oven under vacuum, then ground well. Ground Lion’s foot was dipping in ethanol 80% (1:100 w/v) or in distilled water (1:100 w/v) in dark bottle for 48h in refrigerator at 4˚C temperature. To obtain extracts, then mixtures were filtered by filter paper (Whatman1). Aqueous and ethanolic extracts were evaporated in rotary evaporator at 50˚C.
1) Diet composition: Basal diet was prepared according to Reeves et al. [
2) Experimental design: Animal house in Food Technology Research Institute, Agriculture Research Center albino rats were adapted for one week prior to commencement of the experiment, housed in well aerated cages under hygienic condition and aqueous was introduced ad-libitum. After this week, rats were divided into 8 main groups (seven rats of each) and fed on diets for four weeks as follows: Group 1: Negative control group fed on basal diet. Forty nine rats fed on basal diet and treated with CCl4, in paraffin oil (50% v/v 2 ml/Kg) twice weeks subcutaneous injection to induce chronic damage in the liver [
3) Blood Sampling: At the end of the experiment period, the rats were fasted overnight then the rats were anaesthetized, sacrificed and blood samples were collected from the aorta. The blood samples were centrifuged for 15 minutes at 3000 rpm to separate the serum. The serum was carefully separated into dry clean Wassermann tubes by using a Pasteur pipette and kept frozen till analysis at −20˚C.
1) Liver functions: Aspartate amine transaminase (AST), Alanine amine transaminases (ALT), and Alkaline phosphates (ALP) were measured according to the method described by Tietz et al. [
2) Kidney Functions: Uric acid was determined in the serum according to the method described by Fossati et al. [
Liver and kidney tissues stained by Hematoxylin and Eosin according to Bancroft et al. [
Results were expressed as the mean standard deviation ± SD. Data were statistically analyzed for variance “ANOVA” test at P ≤ (0.05) using SPSS statistical software, “version 20” will be used for these calculations [
Total polyphenols, total flavonoids, soluble tannins, saponins and antioxidant activity of dried lion’s foot are recorded in
Compounds | mg/100g |
---|---|
Total polyphenols | 395.65 ± 21.43* |
Total Flavonoids | 183.10 ± 8.70 |
Soluble Tannins | 150.64 ± 10.22 |
Saponins | 296.32 ± 14.51 |
Antioxidant Activity% (DPPH) | 131.74 ± 2.05 |
*Values are means of triplicates ± Standard deviation.
results were reflected to the antioxidant activity, its noticed that the antioxidant activity of dried lion’s foot leaves was high (131.74%). European Pharmacopoeia described the aerial parts of lion’s foot and its chemical content as containing a minimum of 6% tannin, chlorogenic acid and caffeic acid [
Data in
As seen in
Phenolic compounds | ppm |
---|---|
Gallic acid | 85.65 |
4-Amino-benzoic acid | 33.19 |
Protocatchuic acid | 444.43 |
Catechin | 566.53 |
Chlorogenic acid | 70.10 |
Catechol | 580.54 |
Epicatachin | 264.32 |
Caffeine | 59.32 |
P-OH-benzoic acid | 182.68 |
Caffeic acid | 50.46 |
Vanillic acid | 285.94 |
P?coumaric acid | 153.95 |
Iso-ferulic acid | 69.95 |
Reversetrol | 11.97 |
Ellagic acid | 614.16 |
Alpha-coumaric acid | 66.86 |
Benzoic acid | 1084.63 |
3,4,5-methoxy-cinnamic acid | 30.16 |
Coumarin | 27.67 |
Salycilic acid | 479.71 |
Cinnamic acid | 21.32 |
Flavonoids Compounds | ppm |
---|---|
Luteo.6-arbinose 8-glucose | 40.01 |
Luteo.6-glucose 8-arabinose | 3.19 |
Apig. 6-arabinose 8-galactose | 1.31 |
Apig. 6-rhamnose 8-glucose | 15.04 |
Luteolin | 6.42 |
Narengin | 6.64 |
Rutin | 0.44 |
Rosmarinic | 0.30 |
Apig.7-O-neohespiroside | 1.07 |
Kamp.3,7-dirhamoside | 1.93 |
Apig.7-glucose | 0.69 |
Quercetrin | 0.22 |
Quercetin | 0.75 |
---|---|
Kaemp.3-(2-p-comaroyl)glucose | 6.00 |
Naringenin | 0.24 |
Hespirtin | 0.63 |
Kampferol | 0.33 |
Rhamnetin | 0.56 |
Apegnin | 0.14 |
Acacetin | 8.07 |
Feed efficiency ratio (FER) | Feed Intake (g/d) | Body Weight Gain (%) | Parameters Groups |
---|---|---|---|
0.59a ± 0.016 | 18.98a ± 1.38 | 20.22a ± 2.84 | Normal group |
0.22b ± 0.008 | 13.50b ± 1.65 | 11.03c ± 1.94 | Control positive group (PG) |
0.53a ± 0.011 | 18.89a ± 1.19 | 15.00b ± 1.52 | Ethanol extract 50 ppm |
0.56a ± 0.017 | 18.66a ± 1.92 | 16.00b ± 1.62 | Ethanol extract 100 ppm |
0.58a ± 0.009 | 19.30a ± 1.50 | 15.00b ± 1.99 | Aqueous extract 50 ppm |
0.59a ± 0.013 | 19.05a ± 1.30 | 16.13b ± 1.15 | Aqueousextract 100 ppm |
0.55a ± 0.017 | 18.50a ± 1.4 | 16.00b ± 1.70 | Dried lion’s foot 1% |
0.58a ± 0.019 | 19.00a ± 1.19 | 17.18b ± 2.39 | Dried lion’s foot 2% |
Data are presented as mean (n = 7 rats) ± standard deviation, values with different superscripts within the column are significantly difference at P < 0.05, while those with have similar or partially are not significant.
Parameters Groups | AST (IU/L) | ALT (IU/L) | ALP (IU/L) | Albumin (g/dl ) | Total Bilirubin (mg/dl) |
---|---|---|---|---|---|
Normal group | 53.42c ± 5.88 | 24.50c ± 4.00 | 160.57c ± 17.83 | 3.85a ± 0.11 | 1.70b ± 0.09 |
Control positive group (PG) | 66.57a ± 9.24 | 37.50a ± 6.69 | 219.42a ± 29.37 | 2.05b ± 0.48 | 2.18a ± 0.18 |
Ethanol extract 50 ppm | 58.28b ± 9.64 | 29.85b ± 3.09 | 183.28b ± 10.50 | 3.83a ± 0.20 | 1.75b ± 0.17 |
Ethanol extract 100 ppm | 56.57bc ± 8.36 | 24.14c ± 5.10 | 175.14bc ± 15.09 | 3.45a ± 0.08 | 1.75b ± 0.14 |
Aqueous extract 50 ppm | 60.00ab ± 13.85 | 34.42ab ± 6.27 | 195.42ab ± 16.51 | 3.75a ± 0.26 | 2.07a ± 0.19 |
Aqueousextract 100 ppm | 58.28b ± 9.15 | 30.85b ± 5.79 | 186.00b ± 11.92 | 3.39a ± 0.20 | 1.90b ± 0.15 |
Dried lion’s foot 1% | 55.00bc ± 5.90 | 29.71b ± 4.13 | 187.14b ± 15.92 | 3.71a ± 0.35 | 1.82b ± 0.10 |
Dried lion’s foot 2% | 52.70c ± 6.49 | 24.28c ± 4.06 | 180.00b ± 10.69 | 3.82a ± 0.44 | 1.78b ± 0.14 |
Data are presented as mean (n = 7 rats) ± standard deviation, values with different superscripts within the column are significantly difference at P < 0.05, while those with have similar or partially are not significant.
Parameters Groups | mg/dl | ||
---|---|---|---|
Creatinine | Urea | Uric Acid | |
Normal group | 0.65d ± 0.10 | 35.00d ± 2.44 | 1.80d ± 0.25 |
Control positive group (PG) | 1.30a ± 0.02 | 68.20a ± 7.08 | 3.98a ± 0.42 |
PG treated with ethanol extract 50 ppm | 0.78c ± 0.08 | 43.43c ± 4.99 | 2.73bc ± 0.11 |
PG treated with ethanol extract 100 ppm | 0.66d ± 0.04 | 39.52cd ± 3.12 | 2.25c ± 0.25 |
PG treated with aqueous extract 50 ppm | 0.93b ± 0.07 | 62.30ab ± 5.12 | 3.10ab ± 0.19 |
PG treated with aqueous extract 100 ppm | 0.85bc ± 0.10 | 58.60b ± 4.83 | 2.82bc ± 0.94 |
PG fed on Lion’s foot dried 1% | 0.69d ± 0.08 | 40.55cd ± 6.45 | 2.90c ± 0.92 |
PG fed on Lion’s foot dried 2% | 0.61d ± 0.08 | 38.52cd ± 7.47 | 2.29c ± 0.80 |
All results are expressed as mean ± SD. Values in each column which have different letters are significantly different (p < 0.05).
polyphenols, tannins composed of some gallic and mostly ellagic acid, flavonoids such as orientin, quercetin, quercitrin, isoquercetin, vitexin, rutin, hyperoside, with others, including luteolin and pro-anthocyanidins to which the main pharmacological activities of the plant [
Results in
Liver of rat from group 1 revealed the normal histological structure of hepatic lobule (
4 showed slight vacuolar degeneration of hepatocytes (
Microscopically, kidneys of rat from group 1 showed the normal histological structure of renal tubules (
There is evidence of a hepatoprotective activity of A. vulgaris on the polyphenolic and flavonoids compounds of plant leaves, which have potent antioxidant properties.
El-Hadidy, E.M., Refat, O.G., Halaby, M.S., Elmetwaly, E.M. and Omar, A.A. (2018) Effect of Lion’s Foot (Alchemilla vulgaris) on Liver and Renal Functions in Rats Induced by CCl4. Food and Nutrition Sciences, 9, 46-62. https://doi.org/10.4236/fns.2018.91004