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The acetylcholinesterase (AChE) inhibitors are the main drugs for symptomatic treatment of neurodegenerative disorders like Alzheimer’s disease. A recently designed, synthesized and tested hybrid compound between the AChE inhibitor galantamine (GAL) and the antioxidant polyphenol curcumin (CU) showed high AChE inhibition in vitro. Here, we describe tests for acute and short-term toxicity in mice as well as antioxidant tests on brain homogenates measured the levels of malondialdehide (MDA) and glutathione (GSH). Haematological and serum biochemical analyses were also performed. In the acute toxicity tests, the novel AChE inhibitor given orally in mice showed LD50 of 49 mg/kg. The short-term administration of 2.5 and 5 mg/kg did not show toxicity. In the ex vivo tests, the GAL-CU hybrid performed better than GAL and CU themselves. In a dose of 5 mg/kg, it demonstrates 25% reduction in AChE activity, 28% and 73% increase in the levels of MDA and GSH, respectively. No significant changes in blood biochemical data were observed. The GAL-CU hybrid is a novel non-toxic AChE inhibitor with high antioxidant activity which makes it a perspective multitarget drug candidate for treatment of Alzheimer’s disease.

Three new flavonoid glycosides (7-9) named patuletin-3-O-(5″'-О-Е-feruloyl)-β-d-apiofuranosyl(1→2)[β-d-glucopyranosyl (1→6)]-β-d-glucopyranoside (7), spinacetin-3-O-(5″'-О-Е-feruloyl)-β-d-apiofuranosyl (1→2)[β-d-glucopyranosyl(1→6)]-β-d-glucopyranoside (8) and 6-methoxykaempferol-3-O-(5″'-О-Е-feruloyl)-β-d-apiofuranosyl(1→2)[β-d-glucopyranosyl (1→6)]-β-d-glucopyranoside (9) together with six known flavonoid glycosides of patuletin, spinacetin and 6-methoxykaempferol (1-6) were isolated from the aerial parts of C. bonus-henricus and identified with spectroscopic methods (1D and 2D NMR, UV, IR, HRESIMS). The MeOH extract exerts hepatoprotective and antioxidant activities comparable to those of flavonoid complex silymarin in in vitro (60μg/mL) and in vivo (100mg/kg/daily for 7days) models of hepatotoxicity, induced by CCl4. Flavonoids (1-9) (100μM), compared to silybin, significantly reduced the cellular damage caused by CCl4 in rat hepatocytes, preserved cell viability and GSH level, decreased LDH leakage and reduced lipid damage. High concentrations of compounds (1-9) showed marginal or no cytotoxicity on HepG2 cell line. The experiment data suggest that the glycosides of 6-methoxykaempferol, spinacetin and patuletin are a promising and safe class of hepatoprotective agents.

Geigeria alata Benth. & Hook.f. ex Oliv. & Hiern (Asteraceae) is used in Sudanese folk medicine for treatment of diabetes. The study aimed to estimate the acute oral toxicity of trans-3,5-dicaffeoylquinic acid (3,5-diCQA) from G. alata roots and to assess its antihypeglycemic, antioxidant and antihypertensive effects on chemically-induced diabetic spontaneously hypertensive rats (SHRs). The structure of 3,5-diCQA was established by NMR and HRMS spectra. Type 2 diabetes was induced by intraperitoneal injection of streptozotocin. 3,5-diCQA was slightly toxic with LD50 = 2154 mg/kg. At 5 mg/kg 3,5-diCQA reduced significantly (p

The acetylcholinesterase (AChE) inhibitors are the main drugs for symptomatic treatment of neurodegenerative disorders like Alzheimer’s disease. A recently designed, synthesized and tested hybrid compound between the AChE inhibitor galantamine (GAL) and the antioxidant polyphenol curcumin (CU) showed high AChE inhibition in vitro. Here, we describe tests for acute and short-term toxicity in mice as well as antioxidant tests on brain homogenates measured the levels of malondialdehide (MDA) and glutathione (GSH) and in vitro DPPH, ABTS, FRAP and LPO inhibition assays. Hematological and serum biochemical analyses were also performed. In the acute toxicity tests, the novel AChE inhibitor given orally in mice showed LD50 of 49 mg/kg. The short-term administration of 2.5 and 5 mg/kg did not show toxicity. In the ex vivo tests, the GAL-CU hybrid performed better than GAL and CU themselves, in a dose of 5 mg/kg, it demonstrates 25% reduction in AChE activity, as well as a 28% and 73% increase in the levels of MDA and GSH, respectively. No significant changes in blood biochemical data were observed. The antioxidant activity of 4b measured ex vivo was proven in the in vitro tests. In the ABTS assay, 4b showed radical scavenging activity 10 times higher than the positive control butylhydroxy toluol (BHT). The GAL-CU hybrid is a novel non-toxic AChE inhibitor with high antioxidant activity which makes it a prospective multitarget drug candidate for treatment of neurodegenerative disorders.

The aim of this study was to investigate the effect of Asphodeline lutea (L.) Rchb. dry root extract (ALE) administered alone and against carbon tetrachloride (CCl4)-induced liver injury in vitro/in vivo. The dried roots of A. lutea were extracted with 70% ethanol and was characterized with HPLC-UV. Hepatoprotective potential was investigated by in vivo/in vitro assays in Wistar rats as well as antioxidant properties. At concentrations ranging from 10 to 200μg/mL of ALE significant cytotoxic effects on isolated hepatocytes were found. ALE showed some toxicity in Wistar rats discerned by increased ALT (Alanine transaminase), ALP (Alkaline phosphatase) activities and MDA (malondialdehyde) quantity, decreased GSH (reduced glutathione) levels without affecting the activity of the antioxidant enzymes (GPx (Gluthatione peroxidase), GR (Glutathione reductase) and GST (Glutathione-S-transferase activity)). The antioxidant and hepatoprotective potential of ALE was also observed in vitro/in vivo against CCl4-induced liver injury, where ALE normalizes all the examined parameters perturbated by CCl4 administration. In addition, ALE preserved the decreased cytochrome P450 level and EMND (Ethylmorphine-N-Demethylase) activity without affecting AH (Aniline 4-Hydroxylase) activity. ALE is rich in anthraquinones, naphthalenes and caffeic acid. The pro-oxidant effects of ALE could be due to naphthalene and anthraquinone bioactivation pathways involving toxic metabolites.

Diabetes and hypertension are diseases that often coexist, which increases the risk of chronic organ damages and cardiovascular complications.To evaluate the effects of saponarin, isolated from Gypsophila trichotoma Wend, on blood pressure, glycemia, body weight, and liver biochemical parameters related to oxidative stress in diabetic normotensive Wistar Kyoto rats (NTR) and spontaneously hypertensive rats (SHR).Diabetes was induced by administration of streptozotocin (40 mg/kg, i.p.). The following biochemical parameters: reduced glutathione (GSH), malondialdehyde (MDA), total cytochrome P450, aniline hydroxylase (AH) activity, as well as the activities of antioxidant enzymes such as glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST) were measured in the livers of euthanized rats.Saponarin exerted slight antihypertensive activity in non-diabetic SHR, judged by 19% (p0.05) decrease of the initial blood pressure. However, such effect was not observed in streptozotocin-induced diabetic SHR (SHR-D). Streptozotocin-induced diabetes was evidenced by 78% (p0.05) and by 171% (p0.05) increase in blood glucose level in NTR and SHR, respectively. In non-diabetic SHR the initial MDA quantity was by 36% (p0.05) higher and the initial GSH levels were by 28% (p0.05) lower in comparison to non-diabetic NTR. Significant decrease in the activities of GPx, GR, and GST was measured in the livers of all diabetic rats. Treatment with saponarin ameliorated the above mentioned liver parameters in both diabetic strains, however its effects were less pronounced in the diabetic SHR group.Taken together our data indicate that diabetes and hypertension in combination are more difficult to be modulated by saponarin.

An ultra-high-performance liquid chromatography-high-resolution mass spectrometry based profiling of a purified MeOH extract (PME) from the roots of Chenopodium bonus-henricus L. (Amaranthaceae) tentatively identified 15 saponins of six sapogenins. The PME exerts hepatoprotective and antioxidant activities comparable to those of flavonoid complex silymarin in in vitro (1 and 10 μg/mL) and in vivo (200 mg/kg/daily for 7 days) models of hepatotoxicity, induced by CCl4. The main constituents of PME, respectively saponins bonushenricoside A (1), 3-O-β-D-glucuronopyranosyl-bayogenin-28-O-β-D-glucopyranosyl ester (2), 3-O-β-D-glucuronopyranosyl-medicagenic acid-28-O-β-D-xylopyranosyl (1→4)-α-L-rhamnopyranosyl(1→2)-α-L-arabinopyranosyl ester (3), 3-O-β-D-glucuronopyranosyl-2β-hydroxygypsogenin-28-O-β-D-glucopyranosyl ester (4), 3-O-α-L-rabinopyranosyl-bayogenin-28-O-β-D-glucopyranosyl ester (6) and bonushenricoside B (8) (3 μg/mL each), compared to silymarin (5 and 50 μg/mL), significantly reduced the cellular damage caused by CCl4 in rat hepatocytes, preserved cell viability and glutathione level, decreased lactate dehydrogenase leakage and reduced lipid damage. The experimental data suggest that the glycosides of phytolaccagenin, bayogenin, medicagenic acid, 2β-hydroxygypsogenin, 2β-hydroxyoleanoic acid and oleanoic acid are a promising and safe class of hepatoprotective agents.

This study investigated the possible antioxidant and neuroprotective effects of alcesefoliside, isolated from Astragalus monspessulanus L., Fabaceae, against carbon tetrachloride (CCl4)-induced brain injury in Wistar rats. Iron sulphate/ascorbic acid lipid peroxidation was induced in rat brain microsomes and pre-incubated with alcesefoliside and silybin. Male rats were treated in vivo with alcesefoliside and with silymarin alone; animals challenged with CCl4; and pre-treated with alcesefoliside or silymarin in respective doses for 7 days, challenged with CCl4, followed by curative treatment (additional 14 days). The activity of acetylcholine esterase and the antioxidant enzymes: superoxide-dismutase, catalase, glutathione-peroxidase, glutathione reductase and glutathione-S-transferase as well as the biomarkers of oxidative stress malondialdehyde and reduced glutathione were measured. The alcesefoliside pre-treatment and consecutive curative treatment normalizes the activity of the antioxidant enzymes as well as levels of malondialdehyde and reduced glutathione. The observed effects on tissue level correlate with the histopathological observations of the brain. They were comparable to the effects of silymarin, used as a positive control. The results showed that alcesefoliside has a neuroprotective effect against CCl4-induced brain toxicity in rats. Keywords: Alcesefoliside, Brain, Carbon tetrachloride, Neuroprotection, Oxidative stress

Aim: To investigate the possible antioxidant and hepatoprotective effects of purified extract obtained from aerial parts of Astragalus glycyphyllos (EAG) using in vivo model of CCl 4 - induced liver damage in male Wistar rats. Study Design: A total of 36 animals were randomly allocated in six experimental groups, each consisted of six animals: Control rats ( Group 1), rats challenged orally with CCl 4 (10 % solution in olive oil) ( Group 2), rats treated for 7 days with silymarin (100 mg.kg - 1 , 0.5 mL/100 g) alone ( Group 3) and challenged with CCl 4 after 7 - day pre - treatment with silymarin ( Group 4); animals in groups 5 and 6 were either treated with EAG (100 mg.kg - 1 , 0.5 mL/100 g) alone or challenged with CCl 4 after 7 - day pre - treatment with the extract.

The aim of the current study was to evaluate the effect of a defatted extract (EAS) and three flavonoids, isolated from Astragalus spruneri Boiss. (Fabaceae) using in vitro/in vivo models of liver injury. The EAS was characterized by HPLC and flavonoids (14 mg/g dw) and saponins (8 mg/g dw) were proved. The flavonoids (ASF1, ASF3 and ASF5) were isolated from the same extract and partially identified by LC-MS. In in vitro models of non-enzyme induced (Fe2+/AA) lipid peroxidation in isolated liver microsomes and CCl4-induced metabolic bioactivation and t-BuOOH-induced oxidative stress in isolated rat hepatocytes, both EAS and the flavonoids exerted similar to silybin (positive control) an antioxidant and cytoprotective activity, discerned by decreased MDA production in the microsomes and by preserved cell viability and GSH levels as well as by decreased LDH activity and MDA quantity in isolated rat hepatocytes. The antioxidant and hepatoprotective effect of EAS has been confirmed in vivo against CCl4-induced liver injury in rats. EAS restored the GSH levels and the activity of the antioxidant enzymes CAT and SOD, affected by CCl4 administration, as well as decreased the production of MDA. The effect of EAS was commensurable with those of silymarin.

Objective To investigate the hepatoprotective potential of n-butanolic extract of Astragalus monspessulanus L. (EAM) against in-vitro/in-vivo carbon tetrachloride (CCl(4))-induced liver damage in rats. Silymarin was used as a positive control. Methods and results The in-vitro experiments were carried out in primary isolated rat hepatocytes first incubated with CCl(4) (86 µmol/l). Hepatic injury was discerned by a decrease in cell viability and cell glutathione (GSH) levels, an increase in lactate dehydrogenase leakage into the medium, and an elevation in malondialdehyde (MDA) quantity. Cell pre-incubation with EAM (1 µg/ml and 10 µg/ml) significantly ameliorated the CCl(4)-induced liver damage. In-vivo rats were challenged orally with CCl(4) (10% solution in olive oil) alone and after 7 days pre-treatment with EAM (100 mg/kg body weight per day, oral gavage). CCl(4) damage was judged by an increased production of MDA, depletion of cell GSH, and a decrease in cell antioxidant defense system. EAM pre-treatment normalizes the activities of the antioxidant enzymes and the levels of GSH and MDA. These data are supported by the histopathological examination. Conclusion These results indicate that EAM has a similar significant protective effect, in vitro and in vivo, against CCl(4) induced hepatotoxicity in rat as silymarin.This may be due to its antioxidant and membrane stabilizing properties.

The hepatoprotective potential of saponarin, isolated fromGypsophila trichotoma, was evaluatedin vitro/in vivousing a hepatotoxicity model of paracetamol-induced liver injury. In freshly isolated rat hepatocytes, paracetamol (100 μmol) led to a significant decrease in cell viability, increased LDH leakage, decreased levels of cellular GSH, and elevated MDA quantity. Saponarin (60–0.006 μg/mL) preincubation, however, significantly ameliorated paracetamol-induced hepatotoxicity in a concentration-dependent manner. The beneficial effect of saponarin was also observedin vivo. Rats were challenged with paracetamol alone (600 mg/kg, i.p.) and after 7-day pretreatment with saponarin (80 mg/kg, oral gavage). Paracetamol toxicity was evidenced by increase in MDA quantity and decrease in cell GSH levels and antioxidant defence system. No changes in phase I enzyme activities of AH and EMND and cytochrome P 450 quantity were detected. Saponarin pretreatment resulted in significant increase in cell antioxidant defence system and GSH levels and decrease in lipid peroxidation. The biochemical changes are in good correlation with the histopathological data. Protective activity of saponarin was similar to the activity of positive control silymarin. On the basis of these results, it can be concluded that saponarin exerts antioxidant and hepatoprotective activity against paracetamol liver injuryin vitro/in vivo.

Ethanol extract from Astragalus glycyphylloides was investigated for possible protective effect on carbon tetrachloride (CCl4)-induced cytotoxicity (model of metabolic bioactivation) (86 μmol/L) in isolated rat hepatocytes, a well-controlled biological model system in vitro with high metabolizing capacity. Hepatocytes isolated by two-step collagenase perfusion were treated with 10 μg/mL and 100 μg/mL of the extract and with silymarin—7 μg/mL and 70 μg/mL. In this model, the extract had statistically significant cytoprotective and antioxidant activity, near to those of silymarin. These data are supported by reports about the protective effects of saponin isolated from Astragalus membranaceus and Astragalus sieversianus on hepatotoxicity induced by CCl4 and Paracetamol in mice.

In this study, the in vivo effects of a purified saponin mixture (PSM), obtained from Astragalus corniculatus Bieb., were investigated using two in vivo hepatotoxicity models based on liver damage caused by paracetamol (PC) and carbon tetrachloride (CCl4 ). The effects of PSM were compared with silymarin. Male Wistar rats were challenged orally with 20% CCl4 or PC (2 g/kg) four days after being pre-treated with PSM (100 mg/kg) or silymarin (200 mg/kg). A significant decrease of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase (LDH) activities and glutathione (GSH) levels and an increase of malondialdehyde (MDA) quantity was observed after CCl4 and PC administration alone. PSM pre-treatment decreased serum transaminases and LDH activities and MDA levels and increased the levels of cell protector GSH. Biotransformation phase I enzymes were also assessed in both models. In the CCl4 hepatotoxicity model, pre-treatment with PSM or silymarin resulted in significantly increased activities of ethylmorphine-N-demethylase and aniline 4-hydroxylase activity and cytochrome P450, compared to the CCl4 only group. Neither silymarin nor PSM influenced PC biotransformation. Our results suggest that PSM, obtained from A. corniculatus, Bieb. showed in vivo hepatoprotective and antioxidant activities against CCl4 and PC-induced liver damage comparable to that of silymarin.

The antioxidant effect of saponarin, which is the main flavone isolated from Gypsophila trichotoma Wend., and its protection against cocaine hepatotoxicity were investigated in male Wistar rats. The animals were treated with cocaine (40 mg/kg i.p.) alone and also after 3 consecutive days of pretreatment with saponarin (80 mg/kg p.o.). After 18 hours the rats were sacrificed by decapitation. The production of thiobarbituric acid reactive substances, reduced glutathione (GSH) and the activity of the following antioxidant enzymes: catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase were assessed in liver homogenate. Administered alone, cocaine induced significant hepatotoxicity manifested with GSH depletion and reduced antioxidant defences. Saponarin pretreatment, however, decreased cocaine toxicity both by increasing GSH levels and antioxidant enzyme activities. The results of this study proved the antioxidant activity of saponarin and its protective effect against cocaine-induced oxidative stress and hepatotoxicity.

This study investigated the hepatoprotective activity of saponarin, isolated from Gypsophila trichotoma Wend., using in vitro/in vivo hepatotoxicity model based on carbone tetrachloride (CCl₄)-induced liver damage in male Wistar rats. The effect of saponarin was compared with those of silymarin. In vitro experiments were carried out in primary isolated rat hepatocytes. Cell incubation with CCl₄ (86 μmol l⁻¹) led to a significant decrease in cell viability, increased LDH leakage, decreased levels of cellular GSH and elevation in MDA quantity. Cell pre-incubation with saponarin (60-0.006 μg/ml) significantly ameliorated CCl₄-induced hepatic damage in a concentration-dependent manner. These results were supported by the following in vivo study. Along with decreased MDA quantity and increased level of cell protector GSH, seven day pre-treatment of rats with saponarin (80 mg/kg bw; p.o.) also prevented CCl₄ (10%, p.o.)-caused oxidative damage by increasing antioxidant enzyme activities (CAT, SOD, GST, GPx, GR). Biotransformation phase I enzymes were also assessed. Administered alone, saponarin decreased EMND and AH activities but not at the same extent as CCl₄ did. However, pre-treatment with saponarin significantly increased enzyme activities in comparison to CCl₄ only group. The observed biochemical changes were consistent with histopathological observations where the hepatoprotective effect of saponarin was comparative to the effects of the known hepatoprotecor silymarin. Our results suggest that saponarin, isolated from Gypsophila trichotoma Wend., showed in vitro and in vivo hepatoprotective and antioxidant activity against CCl₄-induced liver damage.