In vitro and ex vivo approach for anti-urolithiatic potential of bioactive fractions of gokhru with simultaneous HPLC analysis of six major metabolites and their exploration in rat plasma

Context: Tribulus terrestris L. (Zygophyllaceae) fruits have long been used in traditional systems of medicine for the treatment of various urinary diseases including urolithiasis.

Objective: To explore the anti-urolithiatic potential of gokhru and to develop an analytical method for quantitative estimation of metabolites for its quality control.

Materials and methods: Aqueous extract of gokhru fruit was prepared through maceration followed by decoction to produce a mother extract, which was further used for polarity-based fractionations. In vitro and ex vivo anti-urolithiatic activity of mother extract and fractions at different concentration (100–1000?μg/mL) were carried out using aggregation assay in synthetic urine and in rat plasma, however, nucleation assay for 30?min was done using confocal microscopy. A simultaneous HPLC method has been developed for quantification of diosgenin, catechin, rutin, gallic acid, tannic acid and quercetin in mother extract and in fractions.

Results: The extraction resulted in 14.5% of w/w mother extract, however, polarity-based fractionation yielded 2.1, 2.6, 1.5, 1.3 and 6.1% w/w of hexane, toluene, dichloromethane (DCM), n-butanol and water fractions, respectively. In vitro and ex vivo studies showed a significant anti-urolithiatic potential of n-butanol fraction. Further, HPLC analysis revealed significantly (p?n-butanol fraction as compared to others fractions.

Discussion and conclusion: In vitro and ex vivo studies demonstrated potent anti-urolithiatic activity of n-butanol fraction which can be developed as new phytopharmaceuticals for urolithiasis. HPLC method can be used for quality control and pharmacokinetic studies of gokhru.     

Curative treatment with extracts of Bombax ceiba fruit reduces risk of calcium oxalate urolithiasis in rats

Context: Drawbacks of presently available treatments for urolithiasis necessitate finding the treatment of hyperoxaluria specifically aimed at reduction in oxalate excretion. Interestingly, many Indian tribes use Bombax ceiba L. (Bombacaceae) fruits as a traditional medicine for the treatment of urinary stones.

Objective: The present study investigated the efficacy of B. ceiba fruit extracts as curative agents in experimentally induced calcium oxalate urolithiatic rats.

Materials and methods: Calcium oxalate lithiasis was induced in rats by oral administration of 0.75% ethylene glycol for 14 consecutive days. Treatments with aqueous and ethanol extract of B. ceiba fruit (400?mg/kg body weight) was performed in the same manner for further 14 consecutive days. Cystone (750?mg/kg body weight) was used as reference antiurolithiatic drug. The urinary excretion and kidney deposition of offending salt components, and serum biochemical parameters were investigated.

Results: Oral administration of ethylene glycol resulted in hyperoxaluria and increased renal excretion of calcium and phosphate. However, supplementation with aqueous and ethanol extracts of B. ceiba fruit significantly (p?<?0.05) reduced the elevated urinary oxalate, showing a regulatory action on endogenous oxalate synthesis. The increased deposition of stone forming constituents in kidneys of calculogenic rats was also significantly lowered with curative treatment of aqueous and ethanol extract.

Discussion and conclusion: The results indicate that the fruit of B. ceiba is endowed with lithontriptic activity warranting further development for curative treatment of urolithiasis.     

Antilithiatic Effect of Melia azedarach. on Ethylene Glycol–Induced Nephrolithiasis in Rats

Abstract

The effect of the aqueous extract of Melia azedarach. Linn. (Meliaceae) against ethylene glycol–induced nephrolithiasis in male Wistar albino rats is summarized in this study. Lithiasis was induced in rats by administering 0.75% ethylene glycol in drinking water for 28 days and was manifested by high urinary calcium, phosphate, oxalate, and low urinary magnesium content. Simultaneous administration of aqueous extract of Melia azedarach. (AEMA; 250 mg/kg body weight) orally for 28 days along with ethylene glycol (0.75%) reduced urinary calcium, oxalate, phosphate, and elevated urinary magnesium level. It also increased the urine volume, thereby reducing the tendency for crystallization. The histopathological studies confirmed the induction of lithiasis as microcrystal deposition was observed in sections of kidney from animals treated with ethylene glycol. This was reduced, however, after treatment with the extract. These observations enable us to conclude that AEMA is effective against ethylene glycol–induced nephrolithiasis.     

Beneficial effect of Citrus limon peel aqueous methanol extract on experimentally induced urolithic rats

Context: Citrus limon (L.) Burm.f. (Rutaceace) is a commonly available fruit variety with high medicinal and industrial values.

Objective: Lemon peel (LP) extract was studied as a potent preventive and curative agent for experimentally induced hyperoxaluric rats.

Materials and methods: Gas chromatography–mass spectrometry (GC–MS) analyses and toxicity study were performed for aqueous methanol LP extract. Twenty-four Wistar rats were segregated into four groups. Group 1: Control; Group 2: Urolithic (ethylene glycol (EG) – 0.75%); Group 3: Preventive study (EG?+?LP extract administration from 0th to 7th week); Group 4: Curative study (EG?+?LP extract administration from 4th to 7th week). Animals received LP extract daily by oral administration (100?mg/kg body weight) for 7 weeks.

Results and discussion: GC–MS analyses revealed that compound 6 was abundant in the LP extract (32%) followed by compound 1 (～21%). The LD₅₀ value of LP extract was found to be >5000?mg/kg of body weight. Urolithic rats showed significantly higher urinary calcium and oxalate (4.47?±?0.44 and 18.86?±?0.55?mg/24 h, respectively) excretion compared with control and experimental rats. Renal function parameters like urea (84?±?8.5 and 96.1?±?3.6?mg/dL), creatinine (1.92?±?0.27 and 1.52?±?0.22?mg/dL), and urinary protein (2.03?±?0.02 and 2.13?±?0.16?mg/24 h) were also reduced by LP extract (p?<?0.001) and corroborated with tissue analyses (SOD, catalase, and MDA levels) and histological studies in normal and experimental animals. Immunohistochemical staining of THP and NF-κB in urolithic animals showed elevated expression than the control, while LP extract suppressed the expression of these proteins.

Conclusion: In conclusion, lemon peel is effective in curing kidney stone disease and also can be used to prevent the disease and its recurrence.     

Aqueous extract of Boerhaavia diffusa root ameliorates ethylene glycol-induced hyperoxaluric oxidative stress and renal injury in rat kidney

Introduction: Boerhaavia diffusa Linn. (Nyctaginaceae) is widely used in traditional Indian medicines against renal afflictions including calcium oxalate (CaOx) urolithiasis and is known for antioxidant activity.

Objective: The present study was designed to investigate the ameliorating effect of aqueous extract of B. diffusa roots (BDE) in hyperoxaluric oxidative stress and renal cell injury.

Material and methods: In vitro antioxidant activity of BDE was estimated in terms of total phenolic content and 1,1-diphenyl-2-picryl hydrazyl free radical scavenging activity. Wistar albino rats were given 0.75% v/v ethylene glycol in drinking water to induce chronic hyperoxaluria and simultaneously BDE was given to nephrolithiasic treated rats at the dose of 100 and 200?mg/kg b.w. orally for 28 days. Urinary volume, oxalate, serum creatinine, blood urea nitrogen (BUN), malondialdehyde (MDA) and antioxidant enzyme (SOD, CAT, GST, GPx) were evaluated.

Results and discussion: BDE extract was found to posses a high total phenolic content and exhibited significant free radicals scavenging activity. Oxalate excretion significantly increased in hyperoxaluric animals as compared to control which was protected in BDE-treated animals. BDE treatment significantly reduced level of MDA and improved the activity of antioxidant enzymes followed by reduction in BUN and serum creatinine. In addition, BDE reduced the number of CaOx monohydrate crystals in the urine. Histological analysis depicted that BDE treatment inhibited deposition of CaOx crystal and renal cell damage.

Conclusion: The present study reveals that antioxidant activity of BDE significantly protects against hyperoxaluric oxidative stress and renal cell injury in urolithiasis.     

The effect of aqueous extract of Rosa damascena on formaldehyde-induced toxicity in mice testes

Context: Rosa damascena L. (Rosaceae) (RD) essential oil and extracts are commonly used as a flavour in herbal medicine which increase libido. Previous studies have shown inhalation of RD flower’s oil increases libido and causes protective effects in formaldehyde (FA)-induced testicular damage.

Objective: The protective effects of aqueous extract of RD on the male reproductive system of mice were examined following FA-induced damage.

Materials and methods: Forty-eight adult NMRI male mice were randomly assigned to six groups (n?=?8): control (normal saline, 10?mg/kg); RD40 (40?mg/kg, p.o.); FA treated (10?mg/kg of 10%, i.p.) and FA?+?RD treated at 10, 20 and 40?mg/kg (FA?+?RD10), (FA?+?RD20) and (FA?+?RD40), respectively, for 40 days. At the end of treatment regimes, serum testosterone (T) level and the reproductive activity, viz. body/organ weights, testicular structure and sperm characteristics were studied.

Results: Formaldehyde administration significantly decreased serum T level (p?p?p?Discussion and conclusions: We may conclude that RD flower extract can withstand effects of FA in the male reproductive system of mice possibly due to its antioxidative properties.     

Quercetin promotes gastrointestinal motility and mucin secretion in loperamide-induced constipation of SD rats through regulation of the mAChRs downstream signal

Context: Quercetin (QCT) has been known as a potential therapeutic strategy for gastrointestinal diseases because it contributes to the stabilization of mast cells, the prevention of histamine release and modulation of CaCC chloride channel.

Objective: We investigated the laxative effect and action mechanism of QCT in Lop-induced constipation model.

Materials and methods: Constipation of SD rats was induced by subcutaneous injection of loperamide (Lop) (4 mg/kg weight) in 0.5% Tween 20 twice a day for three days. After 24?h, the constipation group was further treated with 1× PBS (Lop?+?Vehicle treated group), 10?mg/kg of QCT (Lop?+?LQCT treated group), 20?mg/kg of QCT (Lop?+?MQCT treated group) or 40?mg/kg QCT (Lop?+?HQCT treated group) at once. At 24?h after QCT treatment, the constipation phenotypes were measured and the transverse colon was collected from SD rats.

Results: The gastrointestinal motility, the number of stools and histological structures were significantly recovered in Lop?+?QCT treated group compared with the Lop?+?Vehicle treated group. Also, above activity of epithelial cells and smooth muscle cells were regulated by the mRNA expression of the muscarinic acetylcholine receptors M2 and M3 (mAChR M2 and M3) and some mediators of their downstream signalling pathway. Finally, laxative effects of QCT on mAChR signalling pathway were significantly inhibited by the treatment of mAChR antagonist in primary smooth muscle of rat intestine cells (pRISMCs).

Conclusions: This study provides the first strong evidence that QCT can be considered an important candidate for improving chronic constipation induced by Lop treatment in animal models.     

GC-MS analysis and hepatoprotective activity of the n-hexane extract of Acrocarpus fraxinifolius leaves against paracetamol-induced hepatotoxicity in male albino rats

Context: In Egypt, the burden of liver diseases is exceptionally high.

Objective: To investigate the components of the n-hexane extract of Acrocarpus fraxinifolius Arn. (Leguminosae) and its hepatoprotective activity against paracetamol (APAP)-induced hepatotoxicity in rats.

Material and methods: TRACE GC ultra gas chromatogaphic spectrometry was used for extract analysis. Thirty albino rats were divided into six groups (five rats in each). Group 1 was the healthy control; Groups 2 and 3 were healthy treated groups (250 and 500?mg/kg b.w. of the extract, respectively) for seven days. Group 4 was hepatotoxicity control (APAP intoxicated group). Groups 5 and 6 received APAP?+?extract 250 and APAP?+?extract 500, respectively.

Results: Chromatographic analysis revealed the presence of 36 components. Major compounds were α-tocopherol (18.23%), labda-8 (20)-13-dien-15-oic acid (13.15%), lupeol (11.93%), phytol (10.95%) and squalene (7.19%). In the acute oral toxicity study, the mortality rates and behavioural signs of toxicity were zero in all groups (doses from 0 to 5?g/kg b.w. of A. fraxinifolius). LD₅₀ was found to be greater than 5?g/kg of the extract. Only the high dose (500?mg/kg b.w.) of extract significantly alleviated the liver relative weight (4.01?±?0.06) and biomarkers, as serum aspartate aminotransferase (62.87?±?1.41), alanine aminotransferase (46.74?±?1.45), alkaline phosphatase (65.96?±?0.74), lipid profiles (180.39?±?3.51), bilirubin profiles (2.30?±?0.06) and hepatic lipid peroxidation (114.20?±?2.06), and increased body weight (11.58?±?0.20), serum protein profile (11.09?±?0.46) and hepatic total antioxidant capacity (23.78?±?0.66) in APAP-induced hepatotoxicity in rats.

Conclusion: Our study proves the antihepatotoxic/antioxidant efficacies of A. fraxinifolius hexane extract.     

Healing effects of sumac (Rhus coriaria) in streptozotocin-induced diabetic rats

Context Sumac [Rhus coriaria L. (RC) (Anacardiaceae)] is used as a folk medicine in the treatment of diabetes in Turkey.

Objective This study investigates the in vivo healing and protective effects of lyophilized extract sumac against streptozotocin (STZ)-induced diabetic complications.

Materials and methods Toxicity test was conducted in three different dosages (250, 500 and 1000?mg/kg of plant extracts, respectively). Six groups of seven rats each were used in experiments. Groups were designed as Normal control, Diabetic (DM), DM?+?AC-20?mg/kg, DM?+?Extract-100?mg/kg, DM?+?Extract 250?mg/kg and DM?+?Extract 500?mg/kg group. Experimental diabetes [50?mg/kg, intraperitoneal (i.p.)] was induced by STZ. The effects of oral administration of the extract for 21 d on the level of serum glucose, insulin, C-peptide, lipid profile (LP), hepatic and renal damage biomarkers (HRDB), diabetic serum biomarkers (DSB), glycosylated haemoglobin (HbA1c), antioxidant defence system constituents (ADSCs), malondialdehyde (MDA) and α-glucosidase activity in small intestine tissue were evaluated.

Results The extract decreased the levels of blood glucose in diabetic groups (an average of 31%). Triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein levels were balanced by plant extract (500?mg/kg) supplementation in the diabetic group. Decreased levels of aspartate aminotransferase (89%), alanine aminotransferase (91%), lactate dehydrogenase (35%), alkaline phosphatase (47%), creatinine (25%) and urea (29%) were detected in plant extract (500?mg/kg) supplemented diabetic group. Additionally, a considerable increase in the HRDB, DSB, LP, MDA and fluctuated ADSC levels were restored in RC-extract supplemented groups.

Conclusion RC lyophilized extract has a healing effect on diabetes and diabetes-related complications.     

Paracetamol-induced nephrotoxicity and oxidative stress in rats: the protective role of Nigella sativa

Context Nigella sativa L. (Ranunculaceae) (NS) is traditionally used to treat many conditions such as inflammation.

Objective This study evaluates the effects of NS seeds ethanol extract in paracetamol-induced acute nephrotoxicity in rats.

Materials and methods Forty-eight female Wistar Albino rats were divided into eight groups: I?=?sham; II?=?sham?+?1000?mg/kg NS; III?=?sham?+?140?mg/kg (N-acetyl cysteine) NAC; IV?=?2?g/kg paracetamol; V?=?2?g/kg paracetamol?+?140?mg/kg NAC; VI, VII and VIII?=?2?g/kg paracetamol?+?250, 500 and 1000?mg/kg NS, respectively. Paracetamol administration (oral) was carried out 1?h after NS and NAC administrations (oral), and all animals were sacrificed 24?h later.

Results Paracetamol administration significantly increased serum urea (88.05?U/L) and creatinine (0.80?U/L) when compared with the sham group (49.80 and 0.31?U/L, respectively). However, serum urea level was reduced to 65.60, 56.00 and 54.18?U/L, with 250, 500 and 1000?mg/kg doses of the extract, respectively. Also, serum creatinine level was reduced to 0.64, 0.57 and 0.52?U/L with 250, 500 and 1000?mg/kg doses of the extract, respectively. NS administration increased superoxide dismutase and glutathione, and decreased malondialdehyde levels in the kidneys. Kidney histopathological examinations showed that NS administration antagonized paracetamol-induced kidney pathological damage.

Discussion and conclusions The results suggest NS has a significant nephroprotective activity on paracetamol-induced nephrotoxicity. It may be suggested that the antiinflammatory and antioxidant effects of NS ethanolic extract originated from different compounds of its black seeds.     

Bergenia ciliata extract prevents ethylene glycol induced histopathological changes in the kidney

The present investigation is an attempt to evaluate the effect of Bergenia ciliata extract on kidney of ethylene glycol induced urolithiasis in adult female Wistar rats. The hydro-alcoholic extract of Bergenia ciliata/standard drug cystone were administrated simultaneously at a dose of 150 and 300 mg/kg body weight/day, p.o. along with ethylene glycol (0.75% v/v) for 28 days. Significant changes were observed in body weight and absolute organ weight of ethylene glycol treated rats. Also histopathological results showed disrupted renal parenchyma, degenerative changes in glomeruli and focal calcification in glomerulo-tubular structures in ethylene glycol treated animals. Administration of Bergenia ciliata extract/cystone along with ethylene glycol showed significant protective effect in body weight and organ weight with few stray areas of calcifications in glomeruli. Moreover, Bergenia ciliata extract shows higher renoprotective index than cystone at the same dose level.     

Evaluation of herbal antimalarial MAMA decoction-amodiaquine combination in murine malaria model

Context: Co-administration of amodiaquine with MAMA decoction (MD), an herbal antimalarial drug comprising the leaves of Mangifera indica L. (Anacardiaceae), Alstonia boonei De Wild (Apocynaceae), Morinda lucida Benth (Rubiaceae) and Azadirachta indica A. Juss (Meliaceae) was investigated. The practice of concurrent administration of herbal medicines with orthodox drugs is currently on the increase globally.

Objective: The study was designed to investigate the possible enhancement of the antimalarial potency as well as possible herb–drug interaction resulting from concurrent administration of MAMA decoction with amodiaquine (AQ).

Materials and methods: Combinations of MD with AQ were investigated in chloroquine (CQ)-sensitive Plasmodium berghei NK 65 in varying oral doses (mg/kg) at: sub-therapeutic [MD30?+?AQ1.25], therapeutic [MD120?+?AQ10] and median effective [MD40?+?AQ3.8], using chemosuppressive and curative antimalarial test models. Secondly, P. berghei ANKA (CQ-resistant)-infected mice were orally treated with MD 120, 240, [MD120?+?AQ10] and [MD240?+?AQ10] mg/kg, using both models. The survival times of mice were monitored for 28 d.

Results: ED₅₀ values of MD and AQ were 48.8 and 4.1?mg/kg, respectively. A total parasite clearance of CQ-sensitive P. berghei NK65 was obtained with the therapeutic combination dose in the curative test giving an enhanced survival time. In CQ-resistant P. berghei ANKA-infected mice, [MD120?+?AQ10] and [MD240?+?AQ10] mg/kg gave comparable activities with AQ (10?mg/kg) in both models.

Conclusion: The therapeutic combination dose gave total parasite clearance of CQ-sensitive P. berghei NK65, whereas none of the doses tested showed notable activity against CQ-resistant P. berghei ANKA.     

Co-administration of walnut (Juglans regia) prevents systemic hypertension induced by long-term use of dexamethasone: a promising strategy for steroid consumers

Context: The long-term consumption of glucocorticoids (GCs) may induce serious adverse effects such as hypertension. There is sufficient evidence related to the benefit of walnuts on the cardiovascular system.

Objective: This study assesses the effect of methanol extract of walnut [Juglans regia L. (Juglandaceae)] on dexamethasone-induced hypertension and the possible mechanisms in Wistar rats.

Material and methods: Animals were randomized into control, kernel extract (100 and 200?mg/kg/d, orally), dexamethasone (0.03?mg/kg/d, subcutaneously), dexamethasone?+?kernel (100 and 200?mg/kg/d, separately), and dexamethasone?+?captopril (25?mg/kg/d, orally) groups. Animals were treated with water, kernel extract or captopril by gavage 4 d before and during 11 d of saline or dexamethasone treatment. On the 16th day, blood pressure (BP) was recorded and blood samples were collected to measure nitric oxide (NO). Animal hearts were frozen for measurement of malondialdehyde (MDA) and glutathione peroxidase (GPX).

Results: Dexamethasone increased the diastolic BP and MDA/GPX ratio in comparison with control group (128?±?7 vs. 105?±?3?mmHg, p?p?p?p?Conclusion: Similar to captopril, walnut extract normalized dexamethasone-induced hypertension. A part of this beneficial effect apparently involves maintaining balance of the redox system and NO production.     

Adjuvant anti-angiogenic therapy enhances chemotherapeutic uptake in a murine model of head and neck cancer*

Abstract

Intratumoural metabolic demands result in excessive angiogenic cytokine release leading to unorganised vasculature. Resultant fluid dynamics oppose blood flow and drug penetration due to a marked increase in interstitial fluid hydrostatic pressure. It is hypothesised that anti-angiogenic therapy may function to ‘prune’ vasculature and lead to improved chemotherapeutic penetration. Subcutaneous, OSC19 tumour bearing mice (n?=?5/dose/agent) were administered varying doses of an anti-mouse VEGFR2 (DC101) or an anti-mouse VEGFR3 (31C1) –3 d, –1 d, 0 d, +1 d and +3 d prior to 200?µg of cetuximab fluorescently labelled with IRDye800CW. Fluorescence imaging of tumours was performed 10 d post cetuximab-IRDye800CW dose to monitor therapeutic uptake. Co-administration of dual anti-angiogenic agents at 50–50%, 75–25% and 25–75% using optimal dose and time (–1 d 10?mg/kg anti-VEGFR2 and –1 d 40?mg/kg anti-VEGFR3) was also evaluated. In order to establish vessel normalisation, NG2 (pericyte marker) and CD31 (endothelial cells) ratios were assessed during immunohistochemical staining of tumour sections. Twenty-mg/kg anti-VEGFR3?+?5?mg/kg anti-VEGFR2 significantly (p?<?.0005) reduced tumour size (–73%) compared to control (59%). The 20?mg/kg anti-VEGFR3?+?5?mg/kg anti-VEGFR2 and 30?mg/kg anti-VEGFR3?+?2.5?mg/kg anti-VEGFR2 significantly (p?<?.0004) improved percent-injected cetuximab-IRDye800CW dose/gram tumour tissue compared to other groups. Adjuvant, dual anti-angiogenic therapy targeting VEGFR2 and VEGFR3 significantly enhances tumour chemotherapeutic uptake compared to control.     

Clinical response to golimumab in rheumatoid arthritis patients who were receiving etanercept or adalimumab: results of a multicenter active treatment study

Objective: Evaluate the efficacy and safety of subcutaneous (SC) golimumab?+?methotrexate (MTX) in patients with active rheumatoid arthritis (RA) despite etanercept?+?MTX or adalimumab?+?MTX therapy and evaluate whether intravenous (IV) golimumab could rescue patients who were nonresponders to SC golimumab.

Methods: In this multicenter, assessor-blinded, active-switch study of patients with RA (n?=?433) with inadequate response to etanercept or adalimumab?+?MTX, patients continued MTX and received open-label SC golimumab 50?mg every 4 weeks through week 12. DAS28-ESR good responders at week 16 continued open-label SC golimumab through week 52 (Group 1); nonresponders were randomized to double-blind golimumab SC 50?mg (Group 2-SC) or IV 2?mg/kg (Group 2-IV). Week 14 ACR20 was the primary endpoint; assessments continued through week 52 and for patients in the voluntary long-term extension through week 76. A major secondary endpoint was the proportions of patients with ACR20 response at week 52 relative to week 16 in Group 2-SC and Group 2-IV.

Results: At week 14, 34.9% (p?n?=?75) achieved an ACR20 (62.7%). In Groups 2-SC (n?=?91) and 2-IV (n?=?184), 13.2% and 9.2% had an ACR20 at week 52 relative to week 16, with no significant difference between the randomized groups; 42.9% and 47.8% achieved DAS28-ESR response relative to week 0. Through week 16, 4.6% of patients had a serious adverse event. No differences in the rates or types of adverse events were observed between SC and IV golimumab from weeks 16 to 52. The trial limitations included a higher than expected discontinuation rate as a result of a programming error.

Conclusion: SC golimumab?+?MTX significantly suppressed disease activity in RA patients with inadequate response to etanercept and/or adalimumab + MTX. Patients randomized to Groups 2-SC and 2-IV had lower response rates than Group 1, with no difference between SC or IV mode of administration. The safety profile with IV golimumab was comparable to that established with SC golimumab.

Trial registration: NCT01004432, EudraCT 2009-010582-23.     

Protective effect of Pycnogenol on cisplatin-induced ototoxicity in rats

Context: Pycnogenol^®, which is French maritime pine bark extract, is a potent antioxidant. It is used in medical conditions caused by oxidative stress. Cisplatin (cis-diamminedichloroplatinum II) is an antineoplastic agent. However, its serious side effects such as ototoxicity limit its usage.

Objective: Antioxidants can be used to prevent ototoxicity. We investigated the effect of Pycnogenol^® on cisplatin-induced ototoxicity.

Materials and methods: Rats were randomly assigned to four groups of five. Distortion product-evoked otoacoustic emissions (DPOAE) test was performed for each rat. The experimental groups were as follows: Control Group, Pycnogenol^® Group: 10?mg/kg Pycnogenol^® intraperitoneally for 7 days, Cisplatin Group: intraperitoneally 15?mg/kg single injection of cisplatin on the fifth day, Cisplatin?+?Pycnogenol^® Group: intraperitoneally 10?mg/kg Pycnogenol^® treatment for 7 days, additionally on the fifth day, 15?mg/kg single injection of cisplatin was given. On the eighth day, DPOAE was re-performed and rats were sacrificed. Apoptosis was evaluated histopathologically.

Results: Mean percentage of apoptotic cells was 1.5, 3, 30 and 11% in organ of Corti and 2, 2, 40, 15% in spiral ganglion neurons in Control Group, Pycnogenol^® Group, Cisplatin Group and Cisplatin?+?Pycnogenol^® Group, respectively. Cisplatin Group and Cisplatin?+?Pycnogenol^® Group were significantly different when compared to Control Group histopathologically both in organ of Corti and spiral ganglion neuron (p?<0.001, p?=?0.019, p?=?0.001, p?=?0.015). DPOAE results showed that Cisplatin?+?Pycnogenol^® Group was significantly different when compared to Cisplatin Group at 3, 6 and 8?kHz (p?<?0.05).

Conclusion: Pycnogenol protected against cisplatin ototoxicity. Also, pycnogenol is not ototoxic.     

Adjuvant activity of ethanol extract of Hippophae rhamnoides leaves with inactivated rabies virus antigen

Context: Hippophae rhamnoides L. (Elaeagnaceae), commonly known as seabuckthorn (SBT), is known for its medicinal and nutritional properties.

Objective: Evaluation of in vivo adjuvant activity of SBT leaf extract (SBTE) with inactivated rabies virus antigen (Rb).

Materials and methods: Swiss albino mice were immunized with aqueous-alcoholic SBTE (100?mg/kg body weight) or algel (aluminium hydroxide gel) with or without Rb (5% v/v). After priming, booster was administered on day 14. Rabies virus neutralizing antibody (RVNA) titers were estimated by rapid fluorescent focus inhibition test in sera samples collected on days 7, 14, 21, 28 and 35. Effect of adjuvant administration on cytotoxic T lymphocytes (CTLs), memory T cells, plasma and CD11c⁺ cells was studied by flow cytometry. In vitro hemolysis was assayed in human RBC.

Results: RVNA titers were significantly enhanced (p?p?+ cells (25.8%) as compared to 9.4% cells in Rb immunized mice, showed 3.2-fold increment in LPS induced IL-1β. No RBC hemolysis was observed with SBTE.

Conclusions: This study demonstrates the potential adjuvant activity of SBTE with Rb by increasing RVNA titers and CTL response.     

Changes in pharmacokinetic profiles of acetaminophen and its glucuronide after pretreatment with combinations of N-acetylcysteine and either glycyrrhizin,silibinin or spironolactone in rat

Abstract

1. The present study was to investigate the effects of giving N-acetylcysteine (NAC) alone and in combination with either glycyrrhizin (GL), silibinin (SIB) or spironolactone (SL) on the plasma pharmacokinetic (PK) profiles, hepatic exposure, biliary excretion and urinary excretion of acetaminophen (APAP) and its major metabolite, acetaminophen glucuronide (AG).

2. Groups of rats (n?=?5) were pretreated with oral doses of either NAC, NAC?+?GL, NAC?+?SIB or NAC?+?SL on five occasions every 12?h. At 1?h, after the last dose, they received APAP (200?mg/kg) by intraperitoneal injection. Blood, bile, liver and urine samples were collected at various times after APAP injection and analyzed for APAP and AG by HPLC. NAC alone and NAC?+?SIB did not significantly change the PK profiles of APAP and AG. In contrast, NAC?+?GL decreased the biliary excretion of APAP and AG leading to accumulation of APAP in the liver and systemic circulation whereas NAC?+?SL [multidrug resistance associated 2 (Mrp2) inducer] increased the biliary excretion of AG and decreased the hepatic exposure to APAP and AG.

3. Our results suggest that Mrp2 inhibitor GL should be discouraged with NAC to treat APAP hepatotoxicity. Such PK drug–drug interactions should be considered in the treatment of APAP-induced liver injury.     

Possible effects of Phillyrea latifolia on weight loss in rats fed a high-energy diet

Context Phillyrea latifolia L. (Oleaceae), commonly found in the Mediterranean region in Turkey, is used as medicinal teas for weight loss and hyperglycaemia in folk medicine.

Objective The study investigated the possible effects of P. latifolia leaves aqueous extract’s on weight loss and biochemical–histological changes in the rats fed a high-energy diet (HED), also isolated and determined the main phenolic compounds.

Materials and methods Twenty-four male Wistar albino rats were divided into four equal groups such as the HED group fed a HED, the PLE group given only the extract of P. latifolia leaves (220?mg/kg), the HED?+?PLE group administrated with the extract of leaves (220?mg/kg) after being fed with HED and a control group fed with standard pellet diet.

Results PLE administration caused a remarkable decrement of body weight in the HED?+?PLE group (p?<?0.05). PLE showed an improved effect on structural integrity and decreased leukocyte infiltration in liver and small intestinal tissues. The blood glucose (117.3?mmol/L), leptin (5.6?ng/mL), total cholesterol (61.8?mg/dL) and LDL (9.3?mmol/L) levels were significantly increased in the HED group. PLE administration in the HED group decreased these levels. The levels of HDL (26.8?mmol/L) in the HED?+?PLE group were higher than both control and HED groups. Chemical composition was investigated and luteolin 7-O-glucoside and chlorogenic acid were determined for the first time in Turkish sample from the EtOAc extract of leaves.

Discussion and conclusion Phillyrea latifolia leaves may have beneficial effects on obesity related cellular problems and may become a good source of antidiabetic medication.     

Amelioration of hyperglycaemia and modulation of antioxidant status by Alcea rosea seeds in alloxan-induced diabetic rats

Context: Alcea rosea L. (Malvaceae) has various medicinal uses including anticancer, anti-inflammatory and analgesic properties. However, there is no report on its antidiabetic activity.

Objective: Alcea rosea seed extracts were evaluated for antihyperglycaemic and antioxidative potential in diabetic rats.

Materials and methods: Single intra-peritoneal injection of alloxan (130?mg/kg b.w.) was used for induction of diabetes in Albino Wistar rats. Antihyperglycaemic and antioxidant activities of methanol and aqueous extracts of Alcea rosea seed (100 and 300?mg/kg b.w.), administered orally on daily basis for 15 days, were assessed in vivo for fasting blood glucose level and antioxidant status of liver and pancreas. Metformin was used as a positive control.

Results: Aqueous and methanol extracts (300?mg/kg b.w.) decreased blood glucose level in diabetic rats by 24% and 46%, respectively. Administration of aqueous and methanol extracts at 300?mg/kg b.w. significantly (p?2O₂ decomposed/min/mg of protein), respectively. Similar results were observed for pancreas.

Discussion and conclusions: Antihyperglycaemic and antioxidative potentials of Alcea rosea seeds suggest its usefulness in management of diabetes and its complications. This is the first report on antidiabetic activity of this plant.