Antioxidant properties of Ferulago angulata and its hepatoprotective effect against N-nitrosodimethylamine-induced oxidative stress in rats

Context: Ferulago angulata (Schlecht.) Boiss. (Apiaceae) (FASB) is used to treat liver diseases and has been used both as food and therapeutics by many cultures for thousands of years because of the natural antioxidant compounds.

Objective: This study determines antioxidant properties of FASB flowers, the levels of minerals and vitamins, and also, evaluates the hepatoprotective effect of flowers against N-nitrosodimethylamine (NDMA) induced on liver tissue by assessing antioxidant enzymes and histopathological parameters in Wistar albino rats.

Materials and methods: In the study, the rats were divided into six groups of ten. Control, untreated animals were given 0.9% NaCl. Rats were intraperitoneally given NDMA (10?mg/kg) for the first 7 days. FASB methanol extract (150 and 300?mg/kg) was administered orally for 21 days.

Results: α-Tocopherol, retinol, ascorbic acid, total antioxidant activity, phenolic and flavonoid contents of FASB were 0.70?±?0.13, 0.29?±?0.03?μg/g, 139.32?±?7.06?μg/100?g, 171.61?±?6.05?mM ascorbic acid/g, 90.47?±?4.11?mg GA/g and 37.39?±?2.85?mg QE/g. DPPH and hydroxyl radical scavenging activity was obtained IC₅₀ 67.34?±?4.14 and 64.87?±?4.68?μg/mL, respectively.

Discussion and conclusion: The results of the study indicated that FASB flowers contain high levels of vitamins, minerals, total antioxidant activity, phenolics and flavonoids. Due to the positive effect on significant changes in antioxidant enzymes of liver tissue and histopathological examination, it is thought that the plant could be used as a hepatoprotective.     

The effects of triptolide on the pharmacokinetics of sorafenib in rats and its potential mechanism

Context: Combining sorafenib with triptolide could inhibit tumour growth with greater efficacy than single-agent treatment. However, their herb–drug interaction remains unknown.

Objective: This study investigates the herb–drug interaction between triptolide and sorafenib.

Materials and methods: The effects of triptolide (10?mg/kg) on the pharmacokinetics of different doses of sorafenib (20, 50 and 100?mg/kg) in rats, and blood samples were collected within 48?h and evaluated using LC-MS/MS. The effects of triptolide on the absorption and metabolism of sorafenib were also investigated using Caco-2 cell monolayer model and rat liver microsome incubation systems.

Results: The results showed that the C_max (low dose: 72.38?±?8.76 versus 49.15?±?5.46?ng/mL; medium dose: 178.65?±?21.05 versus 109.31?±?14.17?ng/mL; high dose: 332.81?±?29.38 versus 230.86?±?9.68?ng/mL) of sorafenib at different doses increased significantly with the pretreatment of triptolide, and while the oral clearance rate of sorafenib decreased. The t_1/2 of sorafenib increased significant (p?Discussion and conclusions: These results indicated that triptolide could change the pharmacokinetic profiles of sorafenib in rats; these effects might be exerted via decreasing the intrinsic clearance rate of sorafenib in rat liver.     

Effects of Danshen tablets on pharmacokinetics of atorvastatin calcium in rats and its potential mechanism

Context: Danshen tablets (DST), an effective traditional Chinese multi-herbal formula, are often combined with atorvastatin calcium (AC) for treating coronary heart disease in the clinic.

Objective: This study investigated the effects of DST on the pharmacokinetics of AC and the potential mechanism.

Materials and methods: The pharmacokinetics of AC (1?mg/kg) with or without pretreatment of DST (100?mg/kg) were investigated using LC-MS/MS. The effects of DST (50?μg/mL) on the metabolic stability of AC were also investigated using rat liver microsome incubation systems.

Results: The results indicated that C_max (23.87?±?4.27 vs. 38.94?±?5.32?ng/mL), AUC_(0–t) (41.01?±?11.32 vs. 77.28?±?12.92?ng h/mL), and t_1/2 (1.91?±?0.18 vs. 2.74?±?0.23?h) decreased significantly (p?t_1/2) of AC was also decreased (25.7?±?5.2 vs. 42.5?±?6.1) with the pretreatment of DST.

Discussion and conclusions: This study indicated that the main components in DST could accelerate the metabolism of AC in rat liver microsomes and change the pharmacokinetic behaviors of AC. So these results showed that the herb-drug interaction between DST and AC might occur when they were co-administered. Therefore, the clinical dose of AC should be adjusted when DST and AC are co-administered.     

Antidiabetic,antioxidant, and TNF-α lowering properties of extract of the traditionally used plant Ixeris gracilis in alloxan-induced diabetic mice

Abstract

Context: Ixeris gracilis DC. Stebbins (Asteraceae) is a plant considered to be medicinal by local communities of Meghalaya, India.

Objective: To evaluate the antidiabetic potential, antioxidant activity, and effect of the 80% methanolic extract of the leaves of Ixeris gracilis on tumor necrosis factor-α (TNF-α) expression.

Materials and methods: Varying doses (250–1000?mg/kg body weight) were administered intraperitoneally to normoglycemic mice and their hypoglycemic properties noted for 24?h; the optimum dose observed was used to evaluate its antihyperglycemic activity and effect on glucose tolerance. In vitro antioxidant activity was analyzed by assessing the DPPH radicals scavenging ability of the extract and the total polyphenols, flavonoid, carbohydrate, and protein contents were determined. Diabetic mice were then subjected to daily intraperitoneal injections of the extract for 12 days after which the antioxidant enzyme activities in the tissues were assayed and serum TNF-α was evaluated by ELISA.

Results: The extract displayed varying hypoglycemic activity. The dose of 250?mg/kg body weight exhibited potent antihyperglycemic activity and improved glucose tolerance. The extract was able to scavenge free radicals (IC₅₀ 57.544?µg/ml) and contained polyphenol (76.269?±?0.204?mg GAE/g dry wt), flavonoid (70.070?±?0.626?mg rutin equivalent/g dry wt), protein (4.368?±?8.916?mg/g dry wt), and carbohydrate (558.189?±?0.002?mg/g dry wt). TNF-α level and overall activity of glutathione peroxidase and superoxide dismutase in the liver, kidney, and brain of extract-treated diabetic mice were improved.

Conclusion: The study supports the inclusion of Ixeris gracilis in the list of plants with antidiabetic potential.     

Effects of glycyrrhizin on the pharmacokinetics of asiatic acid in rats and its potential mechanism

Context: Asiatic acid has been reported to possess a wide range of pharmacological activities.

Objective: This study investigates the effects of glycyrrhizin on the pharmacokinetics of asiatic acid in rats and its potential mechanism.

Materials and methods: The pharmacokinetics of orally administered asiatic acid (20?mg/kg) with or without glycyrrhizin pretreatment (100?mg/kg/day for seven days) were investigated using a LC–MS method. Additionally, the Caco-2 cell transwell model and rat liver microsome incubation systems were used to investigate the potential mechanism of glycyrrhizin’s effects on the pharmacokinetics of asiatic acid.

Results: The results showed that the C_max (221.33?±?21.06 vs. 324.67?±?28.64?ng/mL), AUC_0–inf (496.12?±?109.31 vs. 749.15?±?163.95?μg·h/L) and the t_1/2 (1.21?±?0.27 vs. 2.04?±?0.32?h) of asiatic acid decreased significantly (p?p?Discussion and conclusions: In conclusion, these results indicated that glycyrrhizin could decrease the system exposure of asiatic acid, possibly by inducing the activity of P-gp or CYP450 enzyme.     

Influence of andrographolide on the pharmacokinetics of warfarin in rats

Context: Andrographolide and warfarin are often used together in clinics in China. However, the herb-drug interaction between andrographolide and warfarin is still unknown.

Objective: This study investigates the herb-drug interaction between andrographolide and warfarin in vivo and in vitro.

Materials and methods: A sensitive and reliable LC-MS/MS method was developed for the determination of warfarin in male Sprague-Dawley rats plasma, and then the pharmacokinetics of orally administered warfarin (0.5?mg/kg) with or without andrographolide (30?mg/kg/day for 7?days) pretreatment was investigated. In addition, Sprague-Dawley rat liver microsomes incubation systems were used to support the in vivo pharmacokinetic data and investigate its potential mechanism.

Results: The method validation results showed that a sensitive and reliable LC-MS/MS method was developed for the determination of warfarin in rat plasma samples. The pharmacokinetic results indicated that co-administration of andrographolide could increase the systemic exposure of warfarin significantly, including area under the curve (118.92?±?18.08 vs. 60.58?±?9.46?μg?×?h/mL), maximum plasma concentration (3.32?±?0.41 vs. 2.35?±?0.25?μg/mL) and t_1/2 (22.73?±?3.28 vs. 14.27?±?2.67?h). Additionally, the metabolic stability of warfarin increased from 23.5?±?4.7 to 38.7?±?6.1?min with the pretreatment of andrographolide, and the difference was significant (p?Discussion and conclusion: In conclusion, andrographolide could increase the systemic exposure of warfarin in rats when andrographolide and warfarin were co-administered, and possibly by slowing down the metabolism of warfarin in rat liver by inhibiting the activity of CYP3A4 or CYP2C9.     

In vivo antitumour potential of camel’s milk against hepatocellular carcinoma in rats and its improvement of cisplatin renal side effects

Context: Camel milk (CM) is recommended for liver disease patients in Egypt for a strong belief that it has a curative effect.

Objective: The effect of consumption of CM with or without chemotherapeutic drug cisplatin was evaluated on induced hepatocarcinogenesis in rats.

Materials and methods: Wistar male rats (56) were divided into eight groups (7 rats each). Group I was control. Hepatocarcinogenesis was initiated by a single dose of intraperitoneal injection of diethylnitrosamine (DENA) (200?mg/kg BW) and promoted by phenobarbitone (500?ppm) in drinking water in groups V, VI, VII and VIII. Treatment started from 28th till 38th week using CM (5?mL/day) and/or cisplatin (5?mg/kg/3?weeks) in groups II, III IV, VI, VII and VIII. Biochemical analysis, lipid peroxidation and superoxide dismutase (SOD) activity in liver tissue were performed. Histopathology of liver and kidney and immunohistochemistry of placental glutathione-S-transferase (P-GST) in liver were performed and analyzed using image analysis.

Results: Albumin concentration and SOD activity were 3.13?±?0.23 and 311.45?±?41.71 in group VII (DENA &; cisplatin), whereas they were 4.3?±?0.15 and 540.5?±?29.94 in group VII (DENA, CM and cisplatin). The mean area of altered hepatocellular foci and P-GST altered foci decreased in group VI (DENA and CM) (1049.6?±?174.78 and 829.1?±?261) and group VIII (cisplatin and CM) (1615.12?±?436 and 543.9?±?127) compared to group V (DENA only) (4173.74?±?510.7 and 3169.49?±?538.61). Cisplatin caused chronic interstitial nephritis, which was slightly alleviated in group VIII (CM and cisplatin).

Conclusions: CM had an antioxidant effect and together with cisplatin managed to decrease hepatocarcinogenesis.     

Protective effects of Ampelopsis brevipedunculata against in vitro hepatic stellate cells system and thioacetamide-induced liver fibrosis rat model

Context: Ampelopsis brevipedunculata Maxim (Vitaceae) is a traditional medicinal herb used for treating liver disorders.

Objective: The hepatoprotective effects of A. brevipedunculata ethanol extract (ABE) was investigated in experimental models of fibrosis.

Materials and methods: Hepatic stellate cells (HSCs) system in vitro and thioacetamide (TAA)-induced liver fibrosis rat model in vivo were used. Sprague–Dawley rats were divided into five groups of eight each (control, TAA, TAA with ABE 10?mg/kg, ABE 100?mg/kg and silymarin 50?mg/kg groups, respectively). Fibrosis was induced except to the control group by TAA (200?mg/kg, i.p.) twice per week for 13?weeks. ABE and silymarin was administered orally six times per week from the 7th week to the 13th week.

Results: In HSC-T6 cells, ABE (0.1?mg/mL) and silymarin (0.05?mg/mL) significantly (p?p?in vivo studies, ABE (10 and 100?mg/kg) treatment ameliorated the altered levels of serum biomarkers significantly (p?p?p?Conclusion: These results collectively indicate that ABE can potentially be developed as a therapeutic agent in the treatment of hepatic fibrosis.     

Effects of triptolide on pharmacokinetics of amlodipine in rats by using LC–MS/MS

Context: Triptolide and amlodipine are often simultaneously used for reducing urine protein excretion after renal transplantation in China clinics.

Objective: This study investigated the effects of triptolide on the pharmacokinetics of amlodipine in male Sprague–Dawley rats.

Materials and methods: The pharmacokinetics of amlodipine (1?mg/kg) with or without triptolide pre-treatment (2?mg/kg/day for seven?days) were investigated using a sensitive and reliable LC–MS/MS method. Additionally, the inhibitory effects of triptolide on the metabolic stability of amlodipine were investigated using rat liver microsome incubation systems.

Results: The results indicated that when the rats were pre-treated with triptolide, the C_max of amlodipine increased from 13.78?±?3.57 to 19.96?±?4.56?ng/mL (p?T_max increased from 4.04?±?1.15 to 5.89?±?1.64?h (p?AUC_0–t increased by approximately 104% (p?p?Conclusions: In conclusion, these results indicated that triptolide could affect the pharmacokinetics of amlodipine, possibly by inhibiting the metabolism of amlodipine in rat liver when they are co-administered.     

Effects of garlic polysaccharide on alcoholic liver fibrosis and intestinal microflora in mice

Context: Alcoholic liver fibrosis (ALF) is treatable and reversible consequence of liver disease. Intestinal microflora plays an important role in the progression of liver disease. Garlic (Allium sativum L. [Amaryllidaceae]) has been consumed as a traditional medicine to treat liver injury.

Objective: To investigate the effects of garlic polysaccharide (GP) on ALF and intestinal microflora in mice.

Materials and methods: KM mice were orally administered with alcohol (56%, 6?mL/kg) for 30?d to establish ALF model, and divided into four groups together with control group (water only). Hugan tablet (60?mg/kg) or GP (250 and 150?mg/kg) were given 5?h after each dose of alcohol. Biochemical markers in serum and liver homogenate were determined with kits. Alteration of intestinal microflora, and protein expressions of TGF-β1, TNF-α and decorin were detected.

Results: In GP-H group, ALT and AST decreased to 18.85?±?4.71?U/L and 40.84?±?7.89?U/L. MDA, TC, TG and LDL-C decreased to 2.32?±?0.86?mmol/mg, 0.21?±?0.12?mmol/L, 0.96?±?0.31?mmol/L and 0.084?±?0.027?mmol/L. SOD, GSH-Px and GSH increased to 118.32?±?16.32?U/mg, 523.72?±?64.20?U/mg and 0.56?±?0.05?mg/g. Ratios of TGF-β1 and TNF-α decreased to 0.608?±?0.170 and 1.057?±?0.058, decorin increased to 2.182?±?0.129. Lachnospiraceae and Lactobacillus increased, Facklamia and Firmicutes decreased with GP pretreatment.

Discussion and conclusions: Intestinal microflora provides novel insight into the mechanisms of GP that may be used to treat ALF and intestinal microflora dysbiosis.     

Antioxidant,anti-collagenase and anti-elastase activities of Phyllanthus emblica,Manilkara zapota and silymarin: an in vitro comparative study for anti-aging applications

Context Phyllanthus emblica L. (Euphorbiaceae) (amla), Manilkara zapota L.P. Royen (Sapotaceae) (sapota) and silymarin are reported to contain antioxidant effects. However, information on other biological activities relating to the anti-aging properties is limited.

Objective To compare in vitro antioxidants, anti-collagenase (MMP-1 and MMP-2) and anti-elastase properties as well as the phenolic and flavonoid contents of amla, sapota and silymarin as potential anti-aging ingredients.

Materials and methods The ethanol amla and sapota fruit extracts were prepared by three cycles of maceration with 24 h duration each. The total phenolic (TPC) and flavonoid (TFC) contents were determined. The antioxidant capacity was evaluated by DPPH and ABTS assays. The effects of MMP-1, MMP-2 and elastase inhibitions were determined by using the EnzChek® assay kits (Molecular-Probes, Eugene, OR).

Results Amla exhibited the highest in TPC (362.43?±?11.2?mg GAE/g) while silymarin showed the highest in TFC (21.04?±?0.67?mg QE/g). Results of antioxidant activity by DPPH and ABTS methods showed that amla possessed the most potent capacity with IC₅₀ values of 1.70?±?0.07 and 4.45?±?0.10?μg/mL, respectively. Highest inhibitions against MMP-1, MMP-2 and elastase were detected for sapota with IC₅₀ values of 89.61?±?0.96, 86.47?±?3.04 and 35.73?±?0.61?μg/mL, respectively.

Discussion and conclusion Test extracts offered anti-aging properties in different mechanisms. Amla showed the highest phenolic content and antioxidant property with moderate anti-collagenase. Silymarin exhibited measurable flavonoid content with anti-elastase effect. Sapota showed the highest collagenase and elastase inhibitions with moderate antioxidant effect. Thus, extracts might be added as a mixture to gain the overall anti-aging effects.     

The effects of verapamil on the pharmacokinetics of curculigoside in rats

Context: Clarifying the potential mechanism of the poor oral bioavailability of curculigoside would be helpful for for investigating pharmacological effects and clinical applications.

Objective: To clarify the main mechanism for poor oral bioavailability.

Materials and methods: First, the pharmacokinetics of curculigoside (20?mg/kg) in rats with and without pretreatment with verapamil (10?mg/kg) was determined using a sensitive and reliable LC-MS method. Then the effects of verapamil on the transport and metabolic stability of curculigoside were investigated using Caco-2 cell transwell model and rat liver microsome incubation systems.

Results: The results showed that verapamil could significantly increase the peak plasma concentration (from 60.17?ng/mL to 93.66?ng/mL) and AUC_0?t (from 289.57 to 764.02?ng·h/mL) of curculigoside. The Caco-2 cell experiments indicated that the efflux ratio of curculigoside was 3.92 (P_appAB 6.43?±?0.57?×?10?^?7?cm/s; P_appBA 2.52?±?0.37?×?10?^?36?cm/s), P-gp might be involved in the transport of curculigoside, and verapamil could inhibit the efflux of curculigoside and increase the absorption of curculigoside significantly in the Caco-2 cell monolayer. Additionally, the rat liver microsome incubation experiments indicated that verapamil could significantly decrease the intrinsic clearance rate of curculigoside (from 38.8 to 23.6?μL/min/mg protein).

Discussion and conclusion: These results indicated that verapamil could significantly change the pharmacokinetic profiles of curculigoside in rats, the poor absorption due to P-gp mediated efflux in intestine and high intrinsic clearance rate in rat liver may be the main reason for the poor oral absolute bioavailability of curculigoside.     

Treatment of Helicobacter pylori infected mice with Bryophyllum pinnatum,a medicinal plant with antioxidant and antimicrobial properties,reduces bacterial load

Context: Bryophyllum pinnatum (Lam.) Kurz (Crassulaceae) is a plant known for its antiulcer properties.

Objective: This study evaluates the anti-Helicobacter pylori activity of Bryophyllum pinnutum methanol extract with a mouse model and its antioxidant properties.

Materials and methods: Dried leaves of Bryophyllum pinnutum were extracted with methanol and ethyl acetate. Broth microdilution method was used to evaluate the anti-Helicobacter activity of extract samples in vitro. Swiss mice were inoculated with a suspension of Helicobacter pylori and divided into control group and four others that received 125, 250, 500?mg/kg of methanol extract or ciprofloxacin (500?mg/kg), respectively, for 7 days. Helicobacter pylori colonization and bacterial load of mouse stomach was assessed on day 1 and 7 post-treatment. The antioxidant activity of Bryophyllum pinnutum was evaluated through DPPH radical, hydroxyl radical and reducing power assay.

Results: Methanol extract showed a significant anti-Helicobacter activity with MIC and MBC values of 32 and 256?μg/mL, respectively. Bryophyllum pinnatum and ciprofloxacin reduced H. pylori colonization of gastric tissue from 100% to 17%. Bryophyllum pinnatum extract (85.91?±?52.91 CFU) and standard (25.74?±?16.15 CFU) also reduced significantly (p?50 values of 25.31?±?0.34, 55.94?±?0.68 and 11.18?±?0.74?μg/mL, respectively.

Discussion and conclusion: The data suggest that the methanol extract of Bryophyllum pinnatum could inhibit Helicobacter pylori growth, and may also acts as an antioxidant to protect gastric mucosa against reactive oxygen species.     

Effects of Ginkgo leaf tablets on the pharmacokinetics of losartan and its metabolite EXP3174 in rats and its mechanism

Context: Ginkgo leaf tablets (GLTs) and losartan are often simultaneously used for the treatment of hypertension in Chinese clinics. However, the herb–drug interaction between GLT and losartan is still unknown.

Objective: This study investigates the effects of GLT on the pharmacokinetics of losartan and its metabolite EXP3174 in rats and its potential mechanism.

Materials and methods: The pharmacokinetic profiles of losartan and EXP3174 of orally administered losartan (10?mg/kg) with or without GLT pretreatment (80?mg/kg/day for 10?days) in Sprague–Dawley rats were determined. In vitro, the effects of GLT on the metabolic stability of losartan were investigated with rat liver microsomes.

Results: The C_max (1.22?±?0.25 vs 1.85?±?0.37?μg/mL) and the AUC_(0–t) (6.99?±?1.05 vs 11.94?±?1.79?mg·h/L) of losartan increased significantly (p?C_max (1.05?±?0.19 vs 0.72?±?0.12?μg/mL) of EXP3174 decreased significantly (p?t_1/2 of losartan was prolonged significantly from 3.94?±?0.62 to 4.75?±?0.52?h (p?Discussion and conclusions: The results indicate that GLT might increase the plasma concentration of losartan and decrease the concentration of EXP3174 through inhibiting the metabolism of losartan.     

Determination of dihydromyricetin in rat plasma by LC-MS/MS and its application to a pharmacokinetic study

Context: The pharmacokinetics properties of dihydromyricetin (DHM) are still unknown.

Objective: This study investigates the pharmacokinetic characteristics of DHM using a sensitive and reliable LC-MS/MS method.

Materials and methods: A rapid and sensitive LC-MS/MS method was developed for the determination of DHM in male Sprague–Dawley rat plasma. Twelve rats were equally randomized into two groups, including the intravenous group (2?mg/kg) and the oral group (20?mg/kg). Blood samples (250?μL) were collected at designated time points and analyzed using this method. The pharmacokinetic parameters were calculated using DAS 3.0 pharmacokinetic software.

Results: The calibration curve was linear within the range of 0.5–200?ng/mL (r?>?0.998) with the lower limit of quantification at 0.5?ng/mL. After the intravenous injection, DHM reached a maximum concentration of 165.67?±?16.35?ng/mL, and t_1/2 was 2.05?±?0.52?h. However, DHM was not readily absorbed and reached C_max 21.63?±?3.62?ng/mL at approximately 2.67?h following the oral administration of DHM, and t_1/2 was 3.70?±?0.99?h. The MRT for the intravenous group and the oral group were 2.62?±?0.36 and 5.98?±?0.58?h, respectively. The AUC_(0-t) for the intravenous group and the oral group were 410.73?±?78.12 and 164.97?±?41.76?ng·L/mL, respectively, so the absolute bioavailability of DHM was 4.02% which was poor.

Discussion and conclusion: The results indicated that the bioavailability was poor. Further work needs to be conducted to investigate the reason for poor bioavailability and improve this situation.     

The association effect of insulin and clonazepam on oxidative stress in liver of an experimental animal model of diabetes and depression

Abstract

Context: It is known that oxidative stress occurs in peripheral blood in an experimental animal model of diabetes and depression, and acute treatment with insulin and clonazepam (CNZ) has a protective effect on oxidative stress in this model.

Objective: This study evaluated the effect of insulin plus CNZ on oxidative stress parameters in the liver of diabetic male rats induced with streptozotocin (STZ) and subjected to forced swimming test (FST).

Materials and methods: Diabetes was induced by a single intraperitoneal (i.p.) dose of STZ 60?mg/kg in male Wistar rats. Insulin (4?IU/kg) plus CNZ acute i.p. treatment (0.25?mg/kg) was administered 24, 5 and 1?h before the FST. Nondiabetic control rats received i.p. injections of saline (1?mL/kg). Protein oxidative damage was evaluated by carbonyl formation and the antioxidant redox parameters were analyzed by the measurements of enzymatic activities of the superoxide dismutase (SOD), catalase and glyoxalase I (GLO). Glycemia levels also were determined.

Results: Our present study has shown an increase in carbonyl content from diabetic rats subjected to FST (2.04?±?0.55), while the activity of catalase (51.83?±?19.02) and SOD (2.30?±?1.23) were significantly decreased in liver from these animals, which were reverted by the treatment. Also, the activity of GLO (0.15?±?0.02) in the liver of the animals was decreased.

Discussion and conclusion: Our findings showed that insulin plus CNZ acute treatment ameliorate the antioxidant redox parameters and protect against protein oxidative damage in the liver of diabetic rats subjected to FST.     

Effects of rosmarinic acid on acetaminophen-induced hepatotoxicity in male Wistar rats

Context: Drug-induced liver injury is a significant worldwide clinical problem. Rosmarinic acid (RA), a natural phenol, has antioxidant effects.

Objective: The effects of RA against acetaminophen (N-acetyl-p-amino-phenol (APAP))-induced oxidative damage and hepatotoxicity in rats were investigated.

Materials and methods: Male Wistar rats were pretreated with RA (10, 50 and 100?mg/kg, i.g.) for one week. On day 7, rats received APAP (500?mg/kg, i.p.). Then aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total protein, malondialdehyde (MDA), glutathione (GSH), total antioxidant capacity (TAC), glutathione S-transferase (GST), cytochrome CYP450 and histopathological changes were determined.

Results: APAP-induced oxidative stress in liver by a significant increase in the level of MDA (7.6?±?0.21?nmol/mg) as well as a decrease in the contents of TAC (1.75?±?0.14?μmol/g), GSH (1.9?±?0.22?μmol/g) and GST) 3.2?±?0.28?U/mg). RA treatment decreased MDA (4.32?±?0.35?nmol/mg) but increased the contents of TAC (3.51?±?0.34?μmol/g), GSH (3.42?±?0.16?μmol/g) and GST (5.71?±?0.71?μmol/g) in APAP group. RA 100?mg/kg decreased ALT (91.5?±?1.5?U/L), AST (169?±?8.8?U/L) and CYP450 (3?±?0.2?nmol/min/mg) in APAP group. Histologically RA attenuated hepatic damage by decreasing necrosis, inflammation, and haemorrhage in liver sections of APAP group.

Discussion and conclusions: This is the first report that oral administration of RA dose-dependently elicited significant hepatoprotective effects in rats through inhibition of hepatic CYP2E1 activity and lipid peroxidation. RA-protected hepatic GSH and GST reserves and total tissue antioxidant capacity.     

Antidiabetic,antihyperlipidaemic, and antioxidant activity of Syzygium densiflorum fruits in streptozotocin and nicotinamide-induced diabetic rats

Context Syzygium densiflorum Wall. ex Wight & Arn (Myrtaceae) has been traditionally used by local tribes of the Nilgiris, Tamil Nadu, India, for the treatment of diabetes, however, no definitive experimental studies are available.

Objective This study investigates the antidiabetic, antihyperlipidaemic and antioxidant activities of ethanol extract of S. densiflorum (EFSD) fruits in streptozotocin (STZ) and nicotinamide (NA)-induced diabetic rats.

Materials and methods Acute oral toxicity and oral glucose tolerance were assessed in normal rats. The antidiabetic, antihyperlipidaemic and antioxidant activities were investigated in STZ???NA-induced diabetic rats. Diabetic rats were orally administered with glibenclamide (10?mg/kg b.wt), EFSD (200, 400 and 800?mg/kg b.wt) for 28 d. Further, changes in the blood glucose level (BGL), biochemical parameters, antioxidants were observed and histology of pancreas was performed.

Results No toxicity and lethality were observed. Results of the following parameters are represented by treated versus disease control (STZ?+?NA) groups. BGL (161.33?±?22.8 versus 476.17?±?56.58?mg/dl), glycosylated haemoglobin (5.285?±?0.19 versus 8.05?±?0.55%), urea (40.32?±?1.96 versus 75.37?±?2.91?mg/dl), uric acid (1.2?±?0.07 versus 2.16?±?0.05?mg/dl), total cholesterol (89.3?±?5.14 versus 139.7?±?5.95?mg/dl) and triglycerides (79.65?±?2.52 versus 108.9?±?3.61?mg/dl) were significantly decreased, whereas haemoglobin (11.75?±?0.73 versus 7.95?±?0.42?g/dl), high‐density lipoprotein cholesterol (14.2?±?1.11 versus 6.97?±?0.84?mg/dl), total protein (45%) and liver glycogen (87%) were significantly increased in EFSD-treated diabetic group. Significant changes were observed in the enzymatic and non-enzymatic antioxidants in EFSD-treated groups (p?<?0.001). Histopathological examination showed the regeneration of β-cells in Islets of Langerhans.

Conclusion This study confirms the antidiabetic, antihyperlipidaemic and antioxidant activities of S. densiflorum fruits.     

Determination of cepharanthine in rat plasma by LC–MS/MS and its application to a pharmacokinetic study

Context: Cepharanthine (CPA) has been reported to possess a wide range of pharmacological activities.

Objective: This study investigates the pharmacokinetic characteristics after oral or intravenous administration of CPA by using a sensitive and rapid LC–MS/MS method.

Materials and methods: A sensitive and rapid LC–MS/MS method was developed for the determination of CPA in Sprague–Dawley rat plasma. Twelve rats were equally randomized into two groups, including the intravenous group (1?mg/kg) and the oral group (10?mg/kg). Blood samples (250?μL) were collected at designated time points and determined using this method. The pharmacokinetic parameters were calculated.

Results: The calibration curve was linear within the range of 0.1–200?ng/mL (r?=?0.999) with the lower limit of quantification at 0.1?ng/mL. After 1?mg/kg intravenous injection, the concentration of CPA reached a maximum of 153.17?±?16.18?ng/mL and the t_1/2 was 6.76?±?1.21?h. After oral administration of 10?mg/kg of CPA, CPA was not readily absorbed and reached C_max 46.89?±?5.25?ng/mL at approximately 2.67?h. The t_1/2 was 11.02?±?1.32?h. The absolute bioavailability of CPA by oral route was 5.65?±?0.35%, and the bioavailability was poor.

Discussion and conclusions: The results indicate that the bioavailability of CPA was poor in rats, and further research should be conducted to investigate the reason for its poor bioavailability and address this problem.     

Withaferin A protects against spinal cord injury by inhibiting apoptosis and inflammation in mice

Context: Withaferin A (WFA) exhibits diverse pharmaceutical applications on human diseases, including rheumatoid arthritis, cancers and microbial infection.

Objective: We evaluated the neuroprotective role of WFA using a mouse model of spinal cord injury (SCI).

Materials and methods: BALB/c mice were administrated 10?mg/kg of WFA. Gene expression was measured by real-time PCR, western blot and immunohistochemistry. Cell morphology and apoptosis were determined by H&;E staining and TUNEL assay. Motor function was evaluated by the BBB functional scale for continuous 7 weeks.

Results: WFA significantly improved neurobehavioural function and alleviated histological alteration of spinal cord tissues in traumatized mice. Brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) significantly increased in WFA-treated mice. Meanwhile, the expression of Nogo-A and RhoA remarkably decreased in the presence of WFA. Furthermore, the apoptotic cell death was attenuated in mice treated with WFA (31.48?±?2.50% vs. 50.08?±?2.08%) accompanied by decreased bax and increased bcl-2. In addition, WFA decreased the expression of pro-inflammatory mediators such as IL-1β (11.20?±?1.96?ng/mL vs. 17.59?±?1.42?ng/mL) and TNF-α (57.38?±?3.57?pg/mL vs. 95.06?±?9.13?pg/mL). The anti-inflammatory cytokines including TGF-β1 (14.32?±?1.04?pg/mL vs. 9.37?±?1.17?pg/mL) and IL-10 (116.80?±?6.91?pg/mL vs. 72.33?±?9.35?pg/mL) were elevated after WFA administration.

Discussion and conclusion: This study demonstrated that WFA has a neuroprotective role by inhibition of apoptosis and inflammation after SCI in mice.