Modulation of CYP1A1 activity by a Ginkgo biloba extract in the human intestinal Caco-2 cells

Ginkgo biloba is a widely consumed dietary supplement. Some dietary active compounds modulate the activity of biotransformation enzymes inside the enterocytes and more interestingly of cytochrome P-450 1A1 (CYP1A1). This enzyme is of a particular interest because of its implication in the metabolism of some exogenous pro-carcinogens or endogenous molecules. In the present work, we have used Caco-2 cells to study the effect of a standard reference material of a Ginkgo biloba extract (GBE) (10-400 μg/ml), as well as of its major individual active compounds (kaempferol, quercetin, isorhamnetin, ginkgolides and bilobalide), alone or in mixtures, at realistic intestinal concentrations, on the induction of CYP1A1 activity, in the presence or absence of benzo[a]pyrene (B[a]P) (0.1 μg/ml), a well-known CYP1A1 inducer. 3-O-rutinosides of kaempferol, quercetin and isorhamnetin were also tested. We have demonstrated a strong induction (p < 0.005) of CYP1A1 activity and a slight, but significant (p < 0.005), decrease of this activity in the presence of B[a]P by the GBE at the realistic exposure level of 100 μg/ml. The inductive effect was explained, in part, by quercetin and kaempferol after 24 h exposure while unknown compounds seem to be responsible for the strong CYP1A1 induction observed after 6 h exposure. The inhibitory potency of flavonols on CYP1A1 activity in presence of B[a]P was much stronger for the aglycones than for the 3-O-rutinosides, explaining the slight effect observed with the GBE, mainly composed of glycosylated flavonoids. These results indicate that GBEs may disturb intestinal CYP1A1 activity and, in turn, affect the metabolism of other compounds. The present paper thus highlights the necessity to take these side effects into account when administrating Ginkgo biloba herbal supplements.     

Glycine reduces cadmium-induced alterations in the viability and activation of macrophage U937 cells

This study investigates the effect of glycine on cadmium-induced alterations on the viability and activation of the cell line U-937. In this experiment, U-937 cells were pre-treated with 16 μM cadmium (as cadmium chloride). These cadmium-treated cells were later incubated with or without glycine (1–16 μM). After 72 h, it was revealed that glycine significantly (P < 0.05) reduced the tendency of cadmium to reduce the viability of the cells. U-937 cells were also treated with phorbol, 12-myristate, 13-acetate to enhance their transition to the macrophage form. Thereafter, the cells were treated with cadmium with or without glycine (1–16 μM). Twenty-four hours later, the supernatants of each cell culture were assessed for the production of tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin 1 (IL-1), nitric oxide (NO), and catalase activity as indices of the activation of macrophages. The results show that glycine significantly (P < 0.05) reduced the cadmium-induced production of all the markers of the activation of macrophages in a concentration-dependent manner. The findings support the immense antioxidant role of glycine.     

Regioselective sulfonation of dopamine by SULT1A3 in vitro provides a molecular explanation for the preponderance of dopamine-3-O-sulfate in human blood circulation

SULT1A3 is an enzyme that catalyzes the sulfonation of many endogenous and exogenous phenols and catechols. The most important endogenous substrate is dopamine (DA), which is often used as a probe substrate for SULT1A3. We developed a new method for analyzing the SULT1A3 reaction products by high-performance liquid chromatography (HPLC) with electrochemical detection. The sulfonate donor 3′-phosphoadenosine-5′-phosphosulfate (PAPS), DA and the two dopamine sulfates, DA-3-O-sulfate and DA-4-O-sulfate, can be separated within 3 min. This enables quantitation of the sulfates without radioactive PAPS or the precipitation of unreacted PAPS. Both sulfates were synthesized as reference substances and characterized by ¹H and ¹³C nuclear magnetic resonance (NMR), mass spectrometry (MS) and tandem mass spectrometry (MS/MS). The purity of the dopamine sulfates was estimated by HPLC using a diode array detector. We determined the enzyme kinetic parameters for formation of DA-3-O-sulfate and DA-4-O-sulfate using purified recombinant human SULT1A3. The reactions followed Michaelis-Menten kinetics up to 50 μM DA concentration, and strong substrate inhibition was observed at higher concentrations. The apparent K_m values for sulfonation at both hydroxy groups were similar (2.21 ± 0.764 and 2.59 ± 1.06 μM for DA-4-O-sulfate and DA-3-O-sulfate, respectively), but the V_max was approximately six times higher for the formation of the 3-O-sulfate (344 ± 139 nmol/min/mg protein) than the 4-O-sulfate (45.4 ± 16.5 nmol/min/mg protein). These results are in accordance with the observation that DA-3-O-sulfate is more abundant in human blood than DA-4-O-sulfate and that in the crystal structure of SULT1A3 with dopamine bound to the active site, the 3-hydroxy group is aligned to form hydrogen bonds with catalytic residues of the enzyme.     

Quercetin impairs learning and memory in normal mice via suppression of hippocampal phosphorylated cyclic AMP response element-binding protein expression

Quercetin is a naturally occurring dietary flavonol and several reports have shown that quercetin substantially affects cognitive function in disease models, which suggests that quercetin might be a useful agent for treatment of memory dysfunction. However, only one report has examined the effects of quercetin on normal cognitive function. In the present study, we investigated the potential deleterious effects of quercetin on normal cognitive function using Western blot assays and the following behavioral tasks: passive avoidance, Y-maze, and Morris water maze. In the passive avoidance task, pre-acquisition administration of quercetin (10, 20, or 40 mg/kg, p.o.) caused significant cognitive impairments in mice (P < 0.05 or P < 0.01). Quercetin-treated groups (10, 20, or 40 mg/kg, p.o.) also showed significant memory impairments compared with the control group in the Y-maze task (P < 0.05). In the Morris water maze task, there were no significant differences among the groups during training trial sessions, but at the probe trial session, the quercetin-treated group (40 mg/kg, p.o.) spent significantly less time in the target quadrant than did the control group (P < 0.05). In Western blot assays of hippocampal tissue, we found that quercetin-treated groups showed decreased expression of phosphorylated Akt (pAkt), phosphorylated calcium-calmodulin kinase II (pCaMKII), and phosphorylated cyclic AMP response element-binding protein (pCREB). These results suggest that acute administration of quercetin impairs cognitive function by suppression of pAkt and pCaMKII, which, in turn, decreases pCREB expression in the hippocampus.     

A search for hepatoprotective activity of aqueous extract of Rhus coriaria L. against oxidative stress cytotoxicity

The protective effects of different concentrations of aqueous extract of Rhus coriaria L. fruit (75 and 100 μg/ml) and also gallic acid (100 μM) as one of its main components were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. Both extract concentrations and gallic acid (100 μM) significantly (P < 0.05) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) were more effective than gallic acid (100 μM) in protecting hepatocytes against CHP induced lipid peroxidation (P < 0.05). On the other hand gallic acid (100 μM) acted more effective than aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) at preventing hepatocyte membrane lysis (P < 0.05). In addition H₂O₂ scavenging effect of both extract concentrations (75 and 100 μg/ml) were determined in hepatocytes and compared with gallic acid (100 μM). Gallic acid (100 μM) was more effective than aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) at H₂O₂ scavenging activity (P < 0.05).     

Fermentation enhances the in vitro antioxidative effect of onion (Allium cepa) via an increase in quercetin content

Yellow onion (Allium cepa) extract showed enhanced antioxidative effects in 2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC) and 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate and acetyl ester (CM-H₂DCFDA) assay after being treated with a crude enzyme extract from soybean paste fungi, Aspergillus kawachii. HPLC analysis showed two increased and two decreased peaks after enzyme treatment. The decreased peaks were identified as quercetin-3,4′-di-O-β-d-glucoside (1) and quercetin-4′-O-β-d-glucoside (2), and peaks that increased were quercetin-3-O-β-d-glucoside (3) and quercetin (4), respectively. It was expected that 3 and 4 were originated from the glucosidic cleavage of their glucosides, 1 and 2. Among the increased compounds, only quercetin (4) showed strong antioxidative activity in the DPPH assay. In addition, the protective effect against glutamate-induced neurotoxicity in HT22 cells was increased when treated with 25 μg/ml of fermented onion. The enhanced neuroprotective effect was also originated from the increased quercetin content. As a consequence, fermentation raised the quercetin content in onion, and subsequently increased the antioxidative and neuroprotective activities.     

A Penicillium sp. F33 metabolite and its synthetic derivatives inhibit acetyl-CoA:1-O-alkyl-sn-glycero-3-phosphocholine acetyltransferase (a key enzyme in platelet-activating factor biosynthesis) and carrageenan-induced paw edema in mice

Acetyl-CoA:1-O-alkyl-sn-glycero-3-phosphocholine (lyso-PAF) acetyltransferase is a key enzyme in the biosynthesis of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF) in inflammatory cells. Substances which inhibit this enzyme are of therapeutic interest. In this study, we screened for new inhibitors of lyso-PAF acetyltransferase with anti-inflammatory effects. In a metabolite from Penicillium sp. F33, we isolated an acetyltransferase inhibitor identified as dihydrofumigatin (2-methoxy-1,3,4-trihydroxy-5-methylbenzene) from high resolution mass spectrometer and NMR data. Dihydrofumigatin had strong acetyltransferase inhibitory activity, but was not stable in aqueous solution. Thus, we chemically synthesized its oxidized form fumigatin (3-hydroxy-2-methoxy-5-methyl-1,4-benzoquinone) and derivatives thereof, and evaluated their inhibitory effects. Strong inhibitory activity was observed for saturated fatty acid esters of fumigatin; the order of inhibition was 3-decanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone (termed FUD-7, IC₅₀ = 3 μM) > 2-methoxy-5-methyl-3-tetradecanoyloxy-1,4-benzoquinone (termed FUD-8, IC₅₀ = 20 μM) > 3-hexanoyloxy-2-methoxy-5-methyl-1,4-benzoquinone (IC₅₀ = 139 μM). Interestingly, these compounds also significantly suppressed the gene expression of lyso-PAF acetyltransferase/LPCAT2 in mouse bone marrow-derived macrophages stimulated by lipopolysaccharide (LPS). We further evaluated the effect of these substances on anti-inflammatory activity in vivo using the carrageenan-induced mouse paw edema test. FUD-7 and FUD-8 at 2.5 mg/kg showed significant, 47.9–51.7%, inhibition stronger than that of prednisolone at 10 mg/kg (41.9%). These results suggest that FUD-7 and FUD-8 are potent inhibitors with anti-inflammatory activity.     

Alphaxalone, a neurosteroid anaesthetic, increases the activity of the glutamate transporter type 3 expressed in Xenopus oocytes

Glutamate transporters may be important targets for anaesthetic action in the central nervous system. The authors investigated the effects of alphaxalone, an intravenous neurosteroid anaesthetic, on the activity of glutamate transporter type 3 (EAAT3). EAAT3 was expressed in Xenopus oocytes by injecting its mRNA. Two-electrode voltage clamping was used to record membrane currents before, during, and after applying l-glutamate (30 μM) in the presence or absence of alphaxalone. Responses were quantified by integrating current traces and are reported in microCoulombs (μC). Results are presented as means ± S.E.M. l-Glutamate induced inward currents in EAAT3 expressing oocytes, and these currents were dose-dependently increased by alphaxalone. Alphaxalone at 0.01 to 3 μM significantly increased the inward currents. In addition, the treatment of oocytes with phorbol-12-myristate-13-acetate (PMA), a protein kinase C (PKC) activator, significantly increased the transporter currents (1.0 ± 0.2 to 1.4 ± 0.2 μC; P < 0.05). However, treatment with PMA plus alphaxalone did not increase responses further as compared with PMA or alphaxalone alone. Furthermore, pretreatment of oocytes with chelerythrine or staurosporine, two PKC inhibitors, did not affect basal transporter currents, but did significantly reduce alphaxalone-enhanced EAAT3 activity; whereas oocytes pretreated with wortmannin, a phosphatidylinositol 3-kinase (PI3K) inhibitor, showed significant reductions in basal and alphaxalone-enhanced EAAT3 activities. The above results suggest that alphaxalone enhances EAAT3 activity and that PKC and PI3K are involved in this effect.     

Generic systems for the enantioseparation of basic drugs in NACE using single-isomer anionic CDs

The enantioseparation of 10 basic drugs was evaluated in NACE systems using heptakis(2-O-methyl-3-O-acetyl-6-O-sulfo)-β-CD (HMAS-β-CD). For this purpose, a D-optimal design with 21 experimental points was applied. Four antifungal agents (econazole, isoconazole, miconazole, sulconazole), three local anesthetics (bupivacaine, mepivacaine and prilocaine), two sympathomimetics (salbutamol and terbutaline) and one β-blocker (carvedilol) were selected as basic model analytes. The influence on the enantiomeric resolution of anionic CD and BGE anion concentrations as well as the BGE anion nature was investigated. For all studied analytes, the enantiomeric resolution was shown to be significantly influenced by the CD concentration. Based on the observed results, a generic NACE system was recommended, namely 20 mM HMAS-β-CD and 10 mM ammonium camphorSO₃⁻ in methanol acidified with 0.75 M formic acid. Moreover, this NACE system was compared to previous conditions with heptakis(2,3-di-O-methyl-6-O-sulfo)-β-CD (HDMS-β-CD) or heptakis(2,3-di-O-acetyl-6-O-sulfo)-β-CD (HDAS-β-CD). Finally, two generic systems using either HDAS-β-CD or HMAS-β-CD were proposed and evaluated for the enantioseparation of ketamine and norketamine after incubation of ketamine in phenobarbital-induced male rat liver microsomes systems.     

Action of (R)-sila-venlafaxine and reboxetine to antagonize cisplatin-induced acute and delayed emesis in the ferret

The chemotherapeutic drug cisplatin is associated with severe gastrointestinal toxicity that can last for several days. A recent strategy to treat the nausea and emesis includes the combination of a 5-HT₃ receptor antagonist, a glucocorticoid, and an NK₁ receptor antagonist. The present studies explore the use of the selective noradrenaline reuptake inhibitors, (R)-sila-venlafaxine, (R,R)-reboxetine and (S,S)-reboxetine to prevent cisplatin (5 mg/kg, i.p.)-induced acute (0-24 h) and delayed (24-72 h) emesis in ferrets. The positive control regimen of ondansetron and dexamethasone, both at 1 mg/kg/8 h, reduced acute and delayed emesis by 100 (P < 0.001) and 61% (P < 0.05). (R)-sila-venlafaxine at 5 and 15 mg/kg/4 h reduced acute emesis by 86 (P < 0.01) and 66% (P < 0.05), respectively and both enantiomers of reboxetine at 1 mg/kg/12 h also reduced the response by ∼ 70-90% (P < 0.05). Out of the reuptake inhibitors, only (R)-sila-venlafaxine at 15 mg/kg/4 h was active to reduce delayed emesis (a 57% reduction was observed (P < 0.05)); its terminal plasma levels were positively correlated with an inhibition of emesis during the delayed phase (P < 0.05). (R)-sila-venlafaxine was also examined against a higher dose of cisplatin 10 mg/kg, i.p. (3 h test) and it dose-dependently antagonized the response (maximum reduction was 94% at 10 mg/kg, p.o.; P < 0.01) but it was ineffective against apomorphine (0.125 mg/kg, s.c.) and ipecacuanha (2 mg/kg, p.o.)-induced emesis (P > 0.05). In conclusion, the studies provide the first evidence for an anti-emetic potential of noradrenaline reuptake inhibitors to reduce chemotherapy-induced acute and delayed emesis.     

Contents of major bioactive flavones in proprietary traditional Chinese medicine products and reference herb of Radix Scutellariae

A simple and efficient HPLC/UV method for the simultaneous determination of six bioactive flavones, namely baicalein, baicalin, wogonin, wogonoside, oroxylin A and oroxylin A-7-O-glucuronide, has been developed and applied for their content determination in reference herb and proprietary traditional Chinese medicine (PTCM) products of Radix Scutellariae. The chromatographic separation was carried out on a Thermo C₁₈ column and linear gradient elution was employed with a mobile phase containing acetonitrile and 20 mM sodium dihydrogen phosphate buffer (pH 4.6). All the analytes were detected by PDA detector at a wavelength of 270 nm. Contents of the analytes in Radix Scutellariae containing PTCM products in forms of capsule, soft capsule, tablet and dripping pill and the reference herb of Radix Scutellariae were analyzed by sonicator extraction with methanol and water mixture (80:20) containing 1 mM HCl for 30 min followed by HPLC analysis. Separation of the six analytes was achieved within 25 min with good linearity (R² > 0.99). The R.S.D. of both the intra-day and inter-day precision for all the six analytes was below 10.14%. The accuracy at different concentrations was within the range of −7.83 to 4.06%. The extraction recovery was within the range of 89.22–107.33% for all the analytes. Contents of the six flavones were found to vary significantly among different products with glycosides, such as baicalin, wogonoside and oroxylin A-7-O-glucuronide, in much greater quantity than their corresponding aglycones. In addition to baicalin (18.54 ± 0.71%, w/w), the commonly used marker compound for Radix Scutellariae, wogonoside (3.54 ± 0.18%, w/w) and oroxylin A-7-O-glucuronide (2.84 ± 0.14%, w/w) also existed in abundant amount in the reference herb. Our findings suggested that wogonoside and oroxylin A-7-O-glucuronide should also be served as the chemical markers together with baicalin for the quality control of herbs and PTCM products of Radix Scutellariae.     

Simultaneous quantitation of levodopa and 3-O-methyldopa in human plasma by HPLC-ESI-MS/MS: application for a pharmacokinetic study with a levodopa/benserazide formulation

A sensitive and simple method was developed for the quantitation of levodopa and its metabolite 3-O-methyldopa, in human plasma, after oral administration of tablet formulations containing levodopa (200 mg) and benserazide (50 mg). The analytes were extracted by a protein precipitation procedure, using carbidopa as an internal standard. A mobile phase consisting of 0.2% formic acid and acetonitrile (94:6, v/v) was used and chromatographic separation was achieved using ACE C₁₈ column (50 mm × 4.6 mm i.d.; 5 μm particle size). Selected reaction monitoring was performed using the fragmentation transitions m/z 198 → m/z 107, m/z 212 → m/z 166 and m/z 227 → m/z 181 for levodopa, 3-O-methyldopa and carbidopa, respectively. Calibration curves were constructed over the range 50.0-6000.0 ng/mL for levodopa and 25.0-4000.0 ng/mL for 3-O-methyldopa. The method shown to be specific, precise, accurate and provided recovery rates higher than 85% for all analytes. No matrix effect was detected in the samples. The validated method was applied in a pharmacokinetic study with a levodopa/benserazide tablet formulation in healthy volunteers.     

Hydrogen sulfide attenuates lipopolysaccharide-induced cognitive impairment: A pro-inflammatory pathway in rats

The present study investigated the effect of sodium hydrosulfide (NaHS), a H₂S donor, on cognitive impairment and neuroinflammatory changes induced by bilateral intracerebroventricular injections of LPS at a dose of 10 μg/rat. Rats received 5 mg/kg NaHS or volume-matched vehicle administration by intraperitoneal injection 3 days before LPS injection then for 9 days once daily. Morris water maze was used to detect the cognitive function. Compared to the sham-treated rats, LPS injection significantly prolonged the mean escape latency in the navigation test (P < 0.05) and shortened the adjusted escape latency by approximately 30% (P < 0.05). Meanwhile, LPS injection decreased H₂S level but increased pro-inflammatory mediators (i.e., TNF-α, TNFR1, degradation of IκB-α and thereafter activation of NF-κB) in hippocampus. However, these effects of LPS were significantly ameliorated with NaHS treatment (P < 0.05 vs vehicle-treated group). The present data suggest that H₂S attenuates LPS-induced cognitive impairment through reducing the overproduction of pro-inflammatory mediators via inhibition of NF-κB pathways in rats. This study sets the stage for exploring a novel H₂S releasing agent for preventing or retarding the development or progression of neurological disorders such as Alzheimer's disease.     

Respiratory effects of buprenorphine/naloxone alone and in combination with diazepam in naive and tolerant rats

Respiratory depression has been attributed to buprenorphine (BUP) misuse or combination with benzodiazepines. BUP/naloxone (NLX) has been marketed as maintenance treatment, aiming at preventing opiate addicts from self-injecting crushed pills. However, to date, BUP/NLX benefits in comparison to BUP alone remain debated. We investigated the plethysmography effects of BUP/NLX in comparison to BUP/solvent administered by intravenous route in naive and BUP-tolerant Sprague-Dawley rats, and in combination with diazepam (DZP) or its solvent. In naive rats, BUP/NLX in comparison to BUP significantly increased respiratory frequency (f, P < 0.05) without altering minute volume (V_E). In combination to DZP, BUP/NLX significantly increased expiratory time (P < 0.01) and decreased f (P < 0.01), tidal volume (V_T, P < 0.001), and V_E (P < 0.001) while BUP only decreased V_T (P < 0.5). In BUP-tolerant rats, no significant differences in respiratory effects were observed between BUP/NLX and BUP. In contrast, in combination to DZP, BUP/NLX did not significantly alter the plethysmography parameters, while BUP increased inspiratory time (P < 0.001) and decreased f (P < 0.01) and V_E (P < 0.001). In conclusion, differences in respiratory effects between BUP/NLX and BUP are only significant in combination with DZP, with increased depression in naive rats but reduced depression in BUP-tolerant rats. However, BUP/NLX benefits in humans remain to be determined.     

Isolation of phytate from Jatropha curcas kernel meal and effects of isolated phytate on growth, digestive physiology and metabolic changes in Nile tilapia (Oreochromis niloticus L.)

Jatropha curcas seeds are rich in oil and protein. The oil is used for biodiesel production. The defatted Jatropha kernel meal obtained after oil extraction is rich in protein (58-66%) and phytate (9-11%). The phytate rich fraction was isolated from defatted kernel meal using organic solvents (acetone and carbon tetracholride). It had 66% phytate and 22% crude protein. The fingerlings (n = 50, 16.2 ± 0.64 g) were randomly distributed into five groups containing 10 replicates and fed iso-nitrogenous diets (crude protein 36%): control diet containing casein and gelatin as proteins; control diet containing 1.5% and 3% Jatropha phytate (PWP_1.5 and PWP₃, respectively); and control diet containing 1.5% and 3% Jatropha phytate supplemented with phytase (1500 FTU/kg) (PWP_{1.5 + Phytase} and PWP_{3 + Phytase}, respectively). Significantly lower (P < 0.05) growth and feed utilization in PWP_1.5 and PWP₃ groups than for control and both phytase containing groups were observed; whereas feed gain ratio exhibited opposite trend. Protein and lipid digestibilities of the diets, amylase and protease enzyme activities in the intestine were significantly higher (P < 0.05) in PWP_{1.5 + Phytase} and PWP_{3 + Phytase} groups than for PWP_1.5 and PWP₃ groups. Lowest red blood cell counts, and hemoglobin and hematocrit concentrations were observed in PWP₃ group which were not statistically different to those for PWP_1.5 group, but were significantly (P < 0.05) lower than those for all other groups. Highest albumin, globulin and total protein concentrations were observed in PP_{3 + Phytase} group and lowest in PWP_1.5 group; and values for the latter were statistically similar to those for control group. Calcium, phosphorus and glucose concentrations in blood and cholesterol concentration in plasma were significantly lower (P < 0.05) in the phytate enriched groups compared with control and phytase treated groups (PP_{1.5 + Phytase} and PP_{3 + Phytase}). Higher (P < 0.05) alkaline phosphatase activity was observed in phytase supplemented groups compared with that in non-supplemented groups which (PP_{1.5 + Phytase}) was statistically similar to that in control group, whereas alanine transaminase activity in blood exhibited opposite trend. In conclusion, Jatropha phytate present in DJKM is an antinutrient and addition of phytase in the diet containing DJKM is recommended.     

Alleviative effects of s-allyl cysteine and s-ethyl cysteine on MCD diet-induced hepatotoxicity in mice

Alleviative effects of s-allyl cysteine (SAC) and s-ethyl cysteine (SEC) upon methionine and choline deficient (MCD) diet-induced hepatotoxicity in mice were examined. SAC or SEC at 1 g/L was added into drinking water for 7 weeks with MCD diet. MCD feeding significantly increased hepatic triglyceride and cholesterol levels, and elevated the activity of glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme, fatty acid synthase (FAS) and 3-hydroxy-3-methylglutaryl coenzyme A reductase (P < 0.05). However, the intake of SAC or SEC significantly decreased hepatic triglyceride accumulation, and reduced G6PDH and FAS activities (P < 0.05). MCD feeding significantly lowered serum and hepatic glutathione (GSH) levels, increased malondialdehyde (MDA) and oxidized glutathione (GSSG) formation, and suppressed the activity and mRNA expression of glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (P < 0.05). The intake of SAC or SEC significantly increased serum and hepatic GSH levels, decreased MDA and GSSG formation, restored the activity and mRNA expression of GPX, SOD and catalase (P < 0.05). MCD feeding significantly enhanced the mRNA expression of interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, matrix metalloproteinases-9 (MMP-9) and collagen-alpha1 (P < 0.05). The intake of SAC and SEC significantly blunted the mRNA expression of IL-1beta, IL-6, TNF-alpha, TGF-beta1 and collagen-alpha1 (P < 0.05). SEC was greater than SAC in suppressing IL-6 and TNF-alpha expression (P < 0.05), but SAC was greater than SEC in suppressing collagen-alpha1 and TGF-beta1 expression (P < 0.05). These data suggest that SAC and SEC are potent agents against MCD-induced hepatotoxicity.     

Influence of GSTs, CYP2E1 and mEH polymorphisms on 1, 3-butadiene-induced micronucleus frequency in Chinese workers

1,3-butadiene (BD) has been classified as a human carcinogen, however, the relationship between chromosomal damage and its metabolic polymorphisms is not clear. The present study used the CBMN assay to detect chromosomal damage in the peripheral lymphocytes of 166 exposed workers and 41 non-exposed healthy individuals. PCR and PCR-RFLP were applied to detect GSTT1, GSTM1, CYP2E1 c1c2 and mEH Tyr113His, His139Arg polymorphisms. The results demonstrated that the micronucleus (MN) frequency of the exposed workers was significantly higher than controls (P < 0.01). Among the exposed workers, the individuals with high BD exposures are more susceptible to chromosomal damage than those with low exposures (FR = 1.30, 95% CI 1.14-1.53; P < 0.05). Gender-difference was also found in our study: males got lower micronucleus frequency than females. Workers who carried the genotypes of GSTM1 (+), CYP2E1 (c1c2/c2c2) and mEH intermediate (I) group had significantly higher MN frequency than those carrying the genotypes of GSTM1 (−) (FR = 1.29, 95% CI 1.05-1.59; P < 0.05), CYP2E1 (c1c1) (FR = 1.55, 95% CI 1.24-1.93; P < 0.01) or mEH high (H) group (FR = 1.57, 95% CI 1.08-2.34; P < 0.05), respectively. Our data indicated that the current BD exposure level could cause significantly higher MN frequency in workers than controls. Polymorphisms of GSTM1, CYP2E1 and mEH are susceptible to altered chromosome damage.     

Fucoidan, a sulfated polysaccharide from brown algae, against myocardial ischemia-reperfusion injury in rats via regulating the inflammation response

The aim of the study was to determine the effects of fucoidan on rat myocardial ischemia-reperfusion (I/R) model and elucidate the potential mechanisms. Myocardial I/R injury was induced by the occlusion of left anterior descending coronary artery for 30 min followed by reperfusion for 2 h. After 2 h reperfusion, hemodynamics parameters were detected. Blood samples were collected to determine serum levels of tumor necrosis factor-α (TNF-α) and interleukin 6, 10 (IL-6, 10). Hearts were harvested to assess histopathological changes, infarct size (IS), and the content of myeloperoxidase (MPO). The expression of high-mobility group box 1 (HMGB1), phosphor-IκB-α and phosphor-nuclear factor kappa B (NF-κB) were assayed by western blot. Compared with control group, treatment with fucoidan improved left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and the contractility index (P < 0.05, P < 0.01). Fucoidan reduced the myocardial IS, the levels of TNF-α and IL-6, and the activity of MPO (P < 0.05, P < 0.01). Fucoidan down-regulated the expression of HMGB1, phosphor-IκB-α and NF-κB, but increased the content of IL-10 when compared with control (P < 0.05, P < 0.01). Besides, the infiltration of polymorph nuclear leukocytes (PMNs) and histopathological damages in myocardium were decreased in fucoidan treated groups (PMNs, P < 0.05, P < 0.01). These findings revealed that the administration of fucoidan could regulate the inflammation response via HMGB1 and NF-κB inactivation in I/R-induced myocardial damage.     

Inhibitory effect of ethanol extract of Magnolia officinalis and 4-O-methylhonokiol on memory impairment and neuronal toxicity induced by beta-amyloid

The components of the herb Magnolia officinalis have exhibited antioxidant and neuroprotective activities. In this study, we investigated effects of ethanol extract of M.officinalis and its major component 4-O-methylhonokiol on memory dysfunction and neuronal cell damages caused by Aβ. Oral pretreatment of ethanol extract of M. officinalis (2.5, 5 and 10 mg/kg) and 4-O-methylhonokiol (1 mg/kg) into drinking water for 5 weeks suppressed the intraventricular treatment of Aβ_1-42 (0.5 μg/mouse, i.c.v.)-induced memory impairments. In addition, 4-O-methylhonokiol prevented the Aβ_1-42-induced apoptotic cell death as well as β-secretase expression. 4-O-methylhonokiol also inhibited H₂O₂ and Aβ_1-42-induced neurotoxicity in cultured neurons as well as PC12 cells by prevention of the reactive oxygen species generation. 4-O-methylhonokiol also directly inhibited β-secretase activity and Aβ fibrilization in vitro. Thus, ethanol extract of M. officinalis may be useful for prevention of the development or progression of AD, and 4-O-methylhonokiol may be a major active component.     

Quercetin 3-O-methyl ether protects FL83B cells from copper induced oxidative stress through the PI3K/Akt and MAPK/Erk pathway

Quercetin is a bioflavonoid that exhibits several biological functions in vitro and in vivo. Quercetin 3-O-methyl ether (Q3) is a natural product reported to have pharmaceutical activities, including antioxidative and anticancer activities. However, little is known about the mechanism by which it protects cells from oxidative stress. This study was designed to investigate the mechanisms by which Q3 protects against Cu^2 +-induced cytotoxicity. Exposure to Cu^2 + resulted in the death of mouse liver FL83B cells, characterized by apparent apoptotic features, including DNA fragmentation and increased nuclear condensation. Q3 markedly suppressed Cu^2 +-induced apoptosis and mitochondrial dysfunction, characterized by reduced mitochondrial membrane potential, caspase-3 activation, and PARP cleavage, in Cu^2 +-exposed cells. The involvement of PI3K, Akt, Erk, FOXO3A, and Mn-superoxide dismutase (MnSOD) was shown to be critical to the survival of Q3-treated FL83B cells. The liver of both larval and adult zebrafish showed severe damage after exposure to Cu^2 + at a concentration of 5 μM. Hepatic damage induced by Cu^2 + was reduced by cotreatment with Q3. Survival of Cu^2 +-exposed larval zebrafish was significantly increased by cotreatment with 15 μM Q3. Our results indicated that Cu^2 +-induced apoptosis in FL83B cells occurred via the generation of ROS, upregulation and phosphorylation of Erk, overexpression of 14-3-3, inactivation of Akt, and the downregulation of FOXO3A and MnSOD. Hence, these results also demonstrated that Q3 plays a protective role against oxidative damage in zebrafish liver and remarked the potential of Q3 to be used as an antioxidant for hepatocytes.