LC-ESI-MS phenolic contents assessment, antioxidant, and protective ability of Punica granatum root bark extract against ethanol-induced gastric ulcer in rats: in silico H+, K+-ATPase inhibitory pathway study.

The objectives of the current study were to evaluate the Punica granatum root bark extract's (PGE) antioxidant and gastroprotective activities against ethanol-induced gastric ulcers in Wistar rats and to elucidate the putative mechanism of action using in silico analysis. The PGE phytochemical study shows high levels of phenolics, flavonoids, tannins, and polysaccharides. In vitro, the PGE was more effective at scavenging hydroxyl radicals than quercetin and had lower ferric reducing activity than catechin. In vivo, it was revealed that pretreatment of ethanol-ulcerated rats with PGE at oral doses of 100, 200, and 400 mg/kg b.w. offered a dose-dependent shield against ethanol-induced ulcers when compared to Omeprazole (20 mg/kg b.w.) by preventing the development of deep ulcer lesions, lowering gastric juice output and pH rises, boosting gastric mucus production and antioxidant enzyme levels, and attenuating malondialdehyde and myeloperoxidase contents. Moreover, the liquid chromatography-mass spectrometry analysis of PGE identified 5 phenolic acids and 4 flavonoids, which revealed an in silico high oral bioavailability, drug-likenesses, and good binding affinities and thus inhibitory effects on the gastric H+, K+-ATPase enzyme. PGE may have synergistic antioxidant, anti-inflammatory, and H+, K+-proton pump inhibitory actions that contribute to its antiulcer efficacy.

Pistacia atlantica Desf. roots extract: LC-ESI-MS Analysis, antioxidant activity and gastroprotective effect on experimentally-induced ultrastructural gastric ulcers in mice.

Pistacia atlantica Desf. (Atlas Pistachio) is an Anacardiaceae tree traditionally used in Tunisia for the treatment of ophthalmic, stomatitis, and digestive tract diseases. In the present study, the Pistacia atlantica Desf. roots extract (PR) was phytochemically analyzed, for the first time, by LC-ESI-MS for phenolic and flavonoid contents, in vitro tested for its potential antioxidant activity based on the 2.2-diphenyl-1-picrylhydrazyl (DPPH) and the reduced power essays (FRAP), and in vivo tested for its ability to shield against ethanol-induced gastric ulcer in mice. The LC-ESI-MS analysis proved the identification of 12 compounds, including Quinic, Gallic, and Protocatechuic, as major phenolic acids and high levels of flavonoids, such as Catechin, Epicatechin, and Cirsiliol. PR also exhibited a mild in vitro antioxidant activity when compared with ascorbic acid. In vivo pretreatment of ethanol-ulcerated mice with PR doses 50 mg/kg and 100 mg/kg body weight (b.w) significantly reduced (P< .05) gastric lesions at a rate of 20.10% and a rate of 40.90%, respectively, when compared with 60.70% rate of sucralfate (50 mg/kg b.w) evidenced by a dose-dependent manner increase in the gastric mucosa enzymatic (SOD, CAT, GPx) antioxidant levels, the decline of the lipid peroxidation, and the preservation of normal gastric superficial epithelium. The underlying mechanism of PR antiulcerogenic activity could be due to a synergistic effect of phenolic acids and flavonoid contents which enhances the gastric antioxidant defense system.Abbreviations: BHT: butylated hydroxytoluene, b.w: body weight, CAT: catalase, DPPH:1-Diphenyl-2-picrylhydrazyl, DW: dry weight, EtOH: ethanol, FRAP: Ferric reducing antioxidant power, GAE: gallic acid equivalents, GPx: Glutathione peroxidase, QE: quercetin equivalents, LC-ESI-MS: Liquid chromatography-Electrospray Ionization-Tandem Mass Spectrometry, MDA: malondialdehyde, PR: Pistacia root, TBA: thiobarbituric acid reagent, TBARS: thiobarbituric acid reactive substances, TCA: trichloroacetic acid, SOD: Superoxide dismutase.

Protective effects of Mentha spicata against nicotine-induced toxicity in liver and erythrocytes of Wistar rats.

The aim of this study was to investigate the protective effect of Mentha spicata supplementation against nicotine-induced oxidative damage in the liver and erythrocytes of Wistar rats. Bioactive substances were determined by liquid chromatography - electrospray ionization - tandem mass spectrometry analysis. Animals were divided into 4 groups of 6 rats each: a normal control group, a nicotine-treated group (1 mg/kg), a group receiving M. spicata extract (100 mg/kg), and a group receiving both M. spicata extract (100 mg/kg) and nicotine (1 mg/kg). Many phenolic acids were identified in the M. spicata aqueous extract. After 2 months of treatment, nicotine induced an increase in the level of white blood cells and a marked decrease in erythrocytes, hemoglobin, and haematocrit. Aspartate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase activities were also found to be higher in nicotine-treated group than those of the control group. Furthermore, nicotine-treated rats exhibited oxidative stress, as evidenced by a decrease in antioxidant enzymes activities and an increase in lipid peroxidation level in liver and erythrocytes. Interestingly, the oral administration of M. spicata extract by nicotine-treated rats alleviated such disturbances. M. spicata contained bioactive compounds that possess important antioxidant potential and protected liver and erythrocytes against nicotine-induced damage.

Variability of antioxidant and biological activities of Rhus tripartitum related to phenolic compounds.

Rhus species are known in traditional medicine for their therapeutic virtue and their extracts showed numerous important properties including antimalarial, antimicrobial, antiviral, and hypoglycemic and anticonvulsant activities. Rhus tripartitum (Ucria) is a medicinal plant widely used in Tunisia folk medicine against chronic diarrhea and gastric ulcer. This study was designed to examine in vitro and ex vivo antioxidant, anti-inflammatory and anticancer activities of four extracts of Rhus tripartitum root cortex with increasing solvent polarity (hexane, dichloromethane, methanol and water). HPLC was used to identify and quantify phenolic compounds in Rhus extract. Water extract showed the highest antioxidant activity using oxygen radical absorbance capacity (ORAC method) with 8.95 ± 0.47 µmol Trolox/mg and a cell based-assay with 0.28 ± 0.12 µmol Trolox/mg as compared to the other fractions. Moreover, methanol extract displayed the strongest anti-cancer activity against human lung carcinoma (A-549) and colon adenocarcinoma cell lines (DLD-1) with an IC50 value of 60.69 ± 2.58 and 39.83 ± 4.56 µg/ml (resazurin test) and 44.52 ± 5.96 and 55.65 ± 6.00 µg/ml (hoechst test), respectively. Besides, the highest anti-inflammatory activity, inhibiting nitric oxide (NO) release, was exhibited by dichloromethane extract with 31.5 % at 160 µg/ml in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The HPLC analysis showed that catechol and kaempferol were the major phenolics. These data suggest the richness of all fractions of Ucria root on interesting bioactive molecules with different polarity and confirm the known traditional therapeutics virtues of this species for the treatment of dysentery, diarrhea and gastric ulcer.

Phytochemical study and protective effect of Trigonella foenum graecum (Fenugreek seeds) against carbon tetrachloride-induced toxicity in liver and kidney of male rat.

Liver and kidney diseases are a global concern, therefore considerable efforts to obtain fine herbs useful as drugs from medicinal plants are currently in progress. The aim of this work was to study the antioxidant effects of previous supplementation with fenugreek seeds (FS) against carbon tetrachloride (CCl4) toxicity in the liver and kidney. CCl4 toxicity was induced by one dose (i.g. 5ml CCl4/kg of body weight, 50% CCl4 in olive oil) after 7 weeks of normal diet or diet rich in 10% of grinded fenugreek seeds (20g of pellet rat food/rat/day). 24h after the treatment with CCl4, all animals were scarified and biological analyses were performed. A phytochemical study of fenugreek seed extract (FSE) was also carried out. The phytochemical analysis of FS and FSE revealed the presence of polyphenols (5.92±0.02mg EGA/g DM), flavonoids (0.44±0.19mg ER/g DM), polysaccharides and trace elements. DPPH radical-scavenging activity of FSE showed an EC50 of 285.59±2.01µg/ml. In vivo, CCl4 administration significantly (p<0.05) induced an increase liver and kidney biomarkers. A significant (p<0.05) alteration of the antioxidant enzyme activities was also observed. In animals pretreated with FS, the studied parameters were much less shifted. These results indicate that the supplementation with fenugreek seeds is significantly effective in protecting the liver and kidneys from CCl4 toxicity.

Lithium induced, oxidative stress and related damages in testes and heart in male rats: The protective effects of Malva sylvestris extract.

Malva sylvestris is widely used in Mediterranean and European traditional medicine and ethnoveterinary for the treatment of various diseases. This study, carried out on male Wistar rats, evaluates the beneficial effects of Malva sylvestris extract upon lithium carbonate-induced damages in testes and heart. For this purpose, Malva sylvestris extract at a dose of 0.2g/kg was orally administrated, followed by 25mg/kg lithium carbonate (intraperitoneal injection, twice daily). Lithium carbonate treatment significantly (p<0.01) decreased the weight of testes, accessory sex organ and heart, sperm count and motility, and serum testosterone level. In addition, exposure to lithium carbonate significantly (p<0.01) increased lipid peroxidation level (LPO) and decreased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in testes and heart. Treatment with M. sylvestris extract affords substantial protection in testes and heart by altering all the parameters to near normal levels that were further confirmed by histological examination. The beneficial effect of M. Sylvestris extract in several organs could be attributed to the interaction of antioxidant components, such as complex polysaccharides, as confirmed by phytochemical analysis.

Evaluation of the anti-diarrheal activity of the hydromethanolic root extract of Rhus tripartita (Ucria) (Anacardiacae).

INTRODUCTION: Rhus tripartita (Anacardiacae) is a plant which is traditionally used for the treatment of ulcer and diarrhea in Tunisia. However, the scientific basis for this usage has not been well established. The core aim of the present study is to evaluate the antidiarrheal activity of Rhus tripartita root methanolic extract (RRE). MATERIAL AND METHODS: The antidiarrheal activity of RRE oral doses (50, 100, 200 and 300mg/kg) was evaluated using the castor oil-induced diarrhea, the intestinal fluid emptying method and the normal intestinal transit test. The antibacterial activity was tested against four pathogenic bacteria using two methods. The RRE was also phytochemical studied. RESULTS: Diarrhea experiments showed a protective effect of the RRE which produced a significant (p<0.05) and dose-dependent reduction of all the diarrhea parameters. It delayed the onset of diarrhea, produced a significant decrease in the frequency of defecation and the diarrhea score severity and decreased the volume of intestinal fluid induced by castor oil as well as the propulsion intestinal transit. The effect of the extract at the highest dose (300mg/kg) was similar to that of loperamide, the standard anti-diarrheal drug (10mg/kg). The anti-bacterial activity test showed that RRE exhibited a great inhibition activity against four pathogenic bacteria strains (Esherichia coli, Salmonella typhimurium, Salmonella argenosa, Staphylococcus aureus). Oral administration of the extract up to 3g/kg did not produce any acute toxicity in rats. The preliminary phytochemical screening of the RRE revealed the presence of flavonoids, tannins, and polyphenols. CONCLUSION: Results showed that RRE at 300mg/kg possesses the highest anti-diarrheal activity possibly mediated by the inhibitory effects on gastrointestinal propulsion and intestinal fluid accumulation.

Protective effects of vitamins (C and E) and melatonin co-administration on hematological and hepatic functions and oxidative stress in alloxan-induced diabetic rats

The present study aimed to investigate the potential effects of vitamins (C and E)/melatonin co-administration on the hematologic and hepatic functions and oxidative stress in alloxan-induced diabetic rats. The intraperitoneal injection of alloxan (120 mg/kg b.w. for 2 days) induced a significant increase of blood glucose levels (hyperglycemia) associated with serious hematologic disorders (P < 0.01) evidenced by the decrease in the levels of red blood cell count (RBC) (-18 %), hematocrit (Ht) (-18 %), hemoglobin content (Hb) (-36 %), mean corpuscular hemoglobin (MCH) (-17 %), and mean corpuscular hemoglobin concentration (MCHC) (-16 %). The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and the plasmatic levels of total cholesterol and triglyceride contents of diabetic rats were, however, noted to undergo significant increases by 42 % (P < 0.01), 134 % (P < 0.001), 27.5 % (P < 0.01), 147 % (P < 0.001), and 67 % (P < 0.01), respectively, as compared to the control animals. Furthermore, a significant increase in malondialdehyde (MDA) content and a significant decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities were observed in the plasma and hepatic tissues of diabetic rats when compared to the controls. Interestingly, the treatment with vitamins (C, E) in combination with melatonin was noted to reduce the plasma levels of glucose, lower the MDA levels, and restore the hematologic parameters and biochemical and antioxidant levels of diabetic rats back to normal values, alleviating diabetes metabolic disorders in rats

Protective effects of vitamins (C and E) and melatonin co-administration on hematological and hepatic functions and oxidative stress in alloxan-induced diabetic rats.

The present study aimed to investigate the potential effects of vitamins (C and E)/melatonin co-administration on the hematologic and hepatic functions and oxidative stress in alloxan-induced diabetic rats. The intraperitoneal injection of alloxan (120 mg/kg b.w. for 2 days) induced a significant increase of blood glucose levels (hyperglycemia) associated with serious hematologic disorders (P < 0.01) evidenced by the decrease in the levels of red blood cell count (RBC) (-18%), hematocrit (Ht) (-18%), hemoglobin content (Hb) (-36%), mean corpuscular hemoglobin (MCH) (-17%), and mean corpuscular hemoglobin concentration (MCHC) (-16%). The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and the plasmatic levels of total cholesterol and triglyceride contents of diabetic rats were, however, noted to undergo significant increases by 42% (P < 0.01), 134% (P < 0.001), 27.5% (P < 0.01), 147% (P < 0.001), and 67% (P < 0.01), respectively, as compared to the control animals. Furthermore, a significant increase in malondialdehyde (MDA) content and a significant decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities were observed in the plasma and hepatic tissues of diabetic rats when compared to the controls. Interestingly, the treatment with vitamins (C, E) in combination with melatonin was noted to reduce the plasma levels of glucose, lower the MDA levels, and restore the hematologic parameters and biochemical and antioxidant levels of diabetic rats back to normal values, alleviating diabetes metabolic disorders in rats.

Phytochemical, antioxidant and protective effect of Rhus tripartitum root bark extract against ethanol-induced ulcer in rats.

Rhus tripartitum (sumac) is an Anacardiaceae tree with a wide phytotherapeutic application including the use of its roots in the management of gastric ulcer. In the present study the Rhus tripartitum root barks extract (RTE) was phytochemical studied, in vitro tested for their potential antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assay and in vivo evaluated for its ability to prevent ethanol-induced gastric ulcer in rats. The RTE was rich in phenolics, flavonoids, tannins and polysaccharide contents and exhibited a low but not weak in vitro antioxidant activity when compared with (+)-catechin. Pre-treatment with RTE at oral doses 50, 200 and 400 mg/kg body weight was found to provide a dose-dependent protection against ethanol-induced ulcer by averting the deep ulcer lesions of the gastric epithelium, by reducing gastric juice and acid output, by enhancing gastric mucus production by preserving normal antioxidant enzymes activities, and inhibiting the lipid peroxidation. The antiulcerogenic activity of RTE might be due to a possible synergistic antioxidant and antisecretory effects.

Ameliorative effect of Opuntia ficus indica juice on ethanol-induced oxidative stress in rat erythrocytes.

The aim of the present study was to investigate the efficacy of Opuntia ficus indica f. inermis fruit juice (OFIj) on reversing oxidative damages induced by chronic ethanol intake in rat erythrocytes. OFIj was firstly analyzed with HPLC for phenolic and flavonoids content. Secondly, 40 adult male Wistar rats were equally divided into five groups and treated for 90 days as follows: control (C), ethanol-only 3 g/kg body weight (b.w) (E), low dose of OFIj 2 ml/100 g b.w+ethanol (Ldj+E), high dose of OFIj 4 ml/100 g b.w+ethanol (Hdj+E), and only a high dose of OFIj 4 ml/100g b.w (Hdj). HPLC analysis indicated high concentrations of phenolic acids and flavonoids in OFIj. Ethanol treatment markedly decreased the activities of erythrocyte superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), and the level of reduced glutathione (GSH). Changes in the erythrocyte's antioxidant ability were accompanied by enhanced oxidative modification of lipids (increase of malondialdeyde level) and proteins (increase in carbonyl groups). Interestingly, pre-administration of either 2 ml/100 g b.w or 4 ml/100 g b.w of OFIj to ethanol-intoxicated rats significantly reversed decreases in enzymatic as well as non enzymatic antioxidants parameters in erythrocytes. Also, the administration of OFIj significantly protected lipids and proteins against ethanol-induced oxidative modifications in rat erythrocytes. The beneficial effect of OFIj can result from the inhibition of ethanol-induced free radicals chain reactions in rat erythrocytes or from the enhancement of the endogenous antioxidants activities.

Protective effects of fenugreek (Trigonella foenum graecum L.) upon dieldrin-induced toxicity in male rat.

The purpose of this study was to evaluate the protective effects of fenugreek (Trigonella foenum graecum L.) upon dieldrin-induced perturbations of haematological parameters and damages to liver and kidney of male Wistar rats. Under our experimental conditions, dieldrin poisoning resulted in 1) an alteration of several haematological parameters, 2) an oxidative stress evidenced by an increase of lipids peroxidation level associated with an increase of superoxide dismutase activity and a decrease of glutathione peroxidase and catalase activities in hepatic and renal tissues, 3) increased levels of glucose, total cholesterol, triglycerides, creatinine, urea, uric acid and proteins in blood, 4) increased activities of lactate dehydrogenase, alkaline phosphatase and transaminases in blood. Previous administration of fenugreek was found to hinder these dieldrin-induced damages: all hematological, renal and hepatic biomarkers, level of lipids peroxidation and activities of catalase and glutathione-peroxidase in liver and kidney were kept close to control values. This protective effect is mainly attributed to antioxidant properties of fenugreek.

Evaluation of Opuntia ficus indica f. inermis fruit juice hepatoprotective effect upon ethanol toxicity in rats.

The aim of our present study is to investigate the effect of Opuntia ficus indica f. inermis prickly pear juice (PPJ) against ethanol-induced liver injury in rats. Chronic ethanol administration (3 g/kg b.w.) during 90 days to Wistar rats, significantly (p < 0.01) increased the liver lipid and protein oxidation, reduced the glutathione content and the activities of liver antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase and conversely elevated the liver injury biochemical markers like aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyl transferase, lactate dehydrogenase, cholesterol, triglycerides and caused a severe histopathologic injuries. Conversely pre-treatment of ethanol-fed rats with PPJ (20 and 40 ml/kg b.w., orally), interestingly reduced liver lipid and protein oxidation, histopathologic lesions and inhibited the alterations of antioxidant enzymes and the release of biochemical markers. The hepatoprotective effect of PPJ could be due to their capacity to end free radicals chain reactions or to enhance the endogenous antioxidants activities.

Protective effect of Opuntia ficus indica f. inermis prickly pear juice upon ethanol-induced damages in rat erythrocytes.

Juice from the fruit of the cactus Opuntia ficus indica is claimed to possess several health-beneficial properties. The present study was carried out to determine whether O. ficus indica f. inermis fruit extract might have a protective effect upon physiological and morphological damages inflicted to erythrocytes membrane by chronic ethanol poisoning, per os, in rat. Chemical analysis of the extract revealed the presence of polyphenols, flavonoids, ascorbic acid, carotenoids, and betalains. Ethanol administration (3 g/kg b.w, per day for 90 days) induced an increase of malondialdehyde (MDA) and carbonylated proteins levels and a decrease of glutathione (GSH) level in erythrocyte. Ethanol administration also reduced the scavenging activity in plasma and enhanced erythrocytes hemolysis, as compared to control rats. In addition, ethanol intake increased the erythrocyte shape index by +895.5% and decreased the erythrocyte diameter by -61.53% as compared to controls. In animals also given prickly pear juice during the same experimental period, the studied parameters were much less shifted. This protective effect was found to be dose-dependent. It is likely that the beneficial effect of the extract is due to the high content of antioxidant compounds.

Evaluation of antioxidant and antiulcerogenic activities of Opuntia ficus indica f. inermis flowers extract in rats.

The Opuntia ficus indica f. inermis methanolic flowers extract (OMFE) was phytochemical studied, in vitro tested for their potential antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, linoleic acid peroxidation assays and in vivo evaluated for its ability to prevent ethanol-induced gastric ulcer in rats. The OMFE was rich in polysaccharide, phenolics and flavonoids contents and exhibited a moderate in vitro antioxidant activity when compared with (+)-catechin and ascorbic acid. Pre-treatment with OMFE at oral doses 250, 500 and 1000 mg/kg body weight was found to provide a dose-dependent protection against ethanol-induced gastric ulcer by averting the deep necrotic lesions of the gastric epithelium, by preserving normal antioxidant enzymes activities, by inhibiting the lipid peroxidation, the oxidation of protein and the DNA fragmentation in gastric mucosa. The antiulcerogenic activity of OMFE might be due to a possible synergistic antioxidant and antihistaminic-like effects.

Antioxidant and antiulcerogenic activities of Opuntia ficus indica f. inermis root extract in rats.

Opuntia ficus indica f. inermis methanolic root extract (ORE) was investigated for phenolic and flavonoids contents, in vitro evaluated for DPPH radical scavenging activity, reducing power and in vivo tested for its gastro-protective ability against 80% ethanol induced ulcer in rats. Phytochemical test of ORE were positive for phenolic and flavonoid contents. DPPH radical scavenging activity and reducing power of ORE showed an EC(50) of 118.65±2.51 µg/ml and 300 µg/ml respectively. In vivo the pre-treatment of rats with ranitidine (50 mg/kg) and 200, 400, and 800 mg/kg doses of ORE significantly (p<0.05) reduced the 80% ethanol induced-ulcer lesion, with a rate of 82.68%, 49.21%, 83.13%, and 92.59% respectively, and prevented the depletion of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), total glutathione (GSH), and inhibited the increase of myeloperoxidase (MPO) and malondialdehyde (MDA) in rat stomach tissues when compared with ethanol group. Also pre-treatment with ORE marked a dose-dependent attenuation of histopathology changes induced by ethanol. Phenolic and flavonoids wealth, radical scavenging activity, and reducing power, have been implicated for antiulcer property of ORE.