Evaluation of protective effects of diosmin (a citrus flavonoid) in chemical-induced urolithiasis in experimental rats

Context There have not been any conclusive studies of the effects of diosmin, a modified flavanone glycoside obtained from Teucrium gnaphalodes L’Her (Lamiaceae), on urolithiasis.

Objective To evaluate anti-urolithiatic effects of diosmin in ammonium chloride and ethylene glycol-induced renal stone in experimental animals.

Materials and methods Thirty Sprague-Dawley were divided into five groups (n=6) receiving the following treatments, respectively, p.o. for 15 consecutive days: distilled water, 0.75% v/v ethylene glycol?+?2% w/v ammonium chloride, 0.75% v/v ethylene glycol?+?2% w/v ammonium chloride?+?cystone® 750?mg/kg, 0.75% v/v ethylene glycol?+?2% w/v ammonium chloride?+?diosmin 10?mg/kg or 0.75% v/v ethylene glycol?+?2% w/v ammonium chloride?+?diosmin 20?mg/kg. Different biomarkers of urolithiasis in urine and serum were evaluated and histopathological examination of kidney was done.

Results Animals treated with diosmin (both 10 and 20?mg/kg) had significantly (p?<?0.005) decreased in kidney weight, urinary pH, total urinary protein, urinary calcium, phosphorus, serum potassium, sodium, magnesium, creatinine, uric acid and blood urea nitrogen levels and significantly (p?<?0.005) increased in urinary volume, urinary magnesium, potassium, sodium, creatinine, uric acid and serum calcium levels in comparison to animals treated with ethylene glycol and ammonium chloride. However, results were better with diosmin 20?mg/kg in comparison to the control group.

Conclusion Diosmin (10 and 20?mg/kg) has very good anti-urolithiatic activity similar to the standard drug cystone®.     

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.     

Therapeutic and fertility restoration effects of Ionidium suffruticosum on sub-fertile male albino Wistar rats: effects on testis and caudal spermatozoa

Context: Ionidium suffruticosum (L.) Ging (Violaceae) is an important medicinal plant widely used as a herbal traditional medicine in Ayurveda for the treatment of infertility. Currently, little pharmacological information is available on its male fertility properties following prolonged use.

Objective: To investigate I. suffruticosum leaf extracts for male fertility parameters.

Materials and methods: The ethanol lyophilized fraction was administered orally on carbendazim-induced sub-fertility rats (250?mg/kg body weight for 28 days). The effects of fractions on rat’s fertility parameters i.e., body and testes weight, sperm motility, sperm vitality, epididymal sperm counts, its morphology, enzyme and antioxidant stress and histopathology were studied and compared with clomiphene citrate.

Results: The sub-fertile male rats treated with I. suffruticosum leaf extract increased the body weight of 7?g, testis weight of 97?mg, increased cauda epididymal sperm counts of 34.2?×?10⁶ sperm/mL, motility of sperm 46% and vitality 28% also increased and normal sperm morphology also improved up to 32%. The carbendazim-treated group showed loss in body weight of 33?g, testis weight of 851?mg, decreased epididymal sperm counts of 15?×?10⁶ sperm/mL, with sluggish motility and a highly significant fall in the live sperms of about 57%.

Discussion and conclusion: The leaf fraction of I. suffructicosum increased the testicular weight, spermatogenesis, sperm counts, lessened sperm agglutination, and increased testicular oxidative biomarkers, SOD, and CAT. This study therefore supports the usage of I. suffructicosum in traditional medicine for infertility.     

Chrysosplenetin inhibits artemisinin efflux in P-gp-over-expressing Caco-2 cells and reverses P-gp/MDR1 mRNA up-regulated expression induced by artemisinin in mouse small intestine

Context: CYP3A4 and P-gp together form a highly efficient barrier for orally absorbed drugs and always share the same substrates. Our previous work revealed that chrysosplenetin (CHR) significantly augmented the rat plasma level and anti-malarial efficacy of artemisinin (ART), partially due to the uncompetitive inhibition effect of CHR on rat CYP3A. But the impact of CHR on P-gp is still unknown.

Objective: The present study investigates whether CHR interferes with P-gp-mediated efflux of ART and elucidates the underlying mechanism.

Materials and methods: P-gp-over-expressing Caco-2 cells were treated with ART (10?μM) or ART-CHR (1:2, 10:20?μM) in ART bidirectional transport experiment. ART concentration was determined by UHPLC-MS/MS method. Healthy male ICR mice were randomly divided into nine groups (n?=?6) including negative control (0.5% CMC-Na solution, 13?mL/kg), ART alone (40?mg/kg), verapamil (positive control, 40?mg/kg), ART-verapamil (1:1, 40:40?mg/kg), CHR alone (80?mg/kg), ART-CHR (1:0.1, 40:4?mg/kg), ART-CHR (1:1, 40:40?mg/kg), ART-CHR (1:2, 40:80?mg/kg) and ART-CHR (1:4, 40:160?mg/kg). The drugs were administrated intragastrically for seven consecutive days. MDR1 and P-gp expression levels in mice small intestine were examined by performing RT-PCR and western blot analysis. ABC coupling ATPase activity was also determined.

Results: After combined with CHR (1:2), P_app (AP-BL) and P_app (BL-AP) of ART changed to 4.29?×?10?^??⁸ (increased 1.79-fold) and 2.85?×?10?^??^8?cm/s (decreased 1.24-fold) from 2.40?×?10?^??⁸ and 3.54?×?10?^??^8?cm/s, respectively. Efflux ratio (P_BA/P_AB) declined 2.21-fold (p?p?p?Discussion and conclusion: These results confirm that CHR inhibited P-gp activity and reverse the up-regulated P-gp and MDR1 levels induced by ART. It suggested that CHR potentially can be used as a P-gp reversal agent to obstruct ART multidrug resistance.     

Protective effects of caffeoylxanthiazonoside isolated from fruits of Xanthium strumarium on sepsis mice

Abstract

Context: The fruit of Xanthium strumarium L. (Asteraceae) has been used for the treatment of various inflammatory diseases.

Objective: This study investigates the protective effect of caffeoylxanthiazonoside (CYXD) isolated from fruits of X. strumarium on sepsis mice in vitro and in vivo.

Materials and methods: Cecal ligation and puncture (CLP) operation was used to establish the sepsis mice model, and sham mice were also performed. CYXD was administered by intraperitoneal injection (10, 20, and 40?mg/kg/d), then the survival rate was measured in 96?h. Additionally, sepsis mice were induced by injection LPS (2?mg/kg); CYXD was administered by intraperitoneal injection (10, 20, and 40?mg/kg/d), then mice were sacrificed, and serum levels of TNF-α and IL-6 were determined by ELISA assay. Furthermore, the ability of CYXD to neutralize LPS was measured by using the LAL test, and expressions of TNF-α, IL-6 were determined by using real-time fluorogenic PCR.

Results: Results indicated that CYXD significantly elevated survival rates of sepsis mice induced by CLP (p?<?0.05) with survival rates of 35%, 45%, and 65%. Furthermore, the LPS level was decreased obviously by CYXD (1, 2, and 4?mg/L) (p?<?0.05). Additionally, CYXD (10, 20, and 40?mg/kg) can not only significantly decrease TNF-α and IL-6 levels induced by LPS in mice's serum (p?<?0.05), but also inhibit mRNA expressions of TNF-α and IL-6 induced by LPS in RAW 264.7 cells at doses of 20, 40, and 80?μg/mL (p?<?0.05).

Conclusion: Our study demonstrated that CYXD has significant protective effects on sepsis mice.     

Evaluation of effects of repeated sevoflurane exposure on rat testicular tissue and reproductive hormones

Abstract

Context: Evaluation of inhalation anesthetics on sperm and reproductive hormones are extremely important.

Objective: Investigation of the effects of sevoflurane used as an inhalation anesthetic on sperm morphology and reproductive hormones in rat testes.

Materials and methods: Forty Wistar-Albino male rats were divided into five groups of eight rats each. The control group received 2?L/min oxygen for seven days, 2?h/day while sevoflurane treatment S1 received 1 minimal alveolar concentration (MAC) sevoflurane?+?2?L/min oxygen for seven days, 2?h/day, and sevoflurane S2 received 1 MAC sevoflurane?+?2?L/min oxygen for seven days, 2?h/day followed by seven days of no treatment. Sevoflurane treatment S3 received 1 MAC sevoflurane?+?2?L/min oxygen for 14 days, 2?h/day and sevoflurane treatment S4 received 1 MAC sevoflurane?+?2?L/min oxygen for 14 days, 2?h/day, with no treatment for the following seven days. All rats were examined histologically after experimental procedures. Rat luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), and inhibin levels were measured.

Results: Histological injury scores were significantly higher in S2, S3, and S4 receiving sevoflurane in comparison to the control group (p?=?0.001, <0.001, and 0.001, respectively). Sperm motility and concentration decreased in S3 and S4 compared to the control group (p?=?0.03 and 0.02, respectively). Significant differences were detected among all groups for serum LH, FSH, T, and inhibin serum concentrations (p?<?0.05).

Conclusion: Testicular and sperm morphology, and reproductive hormones were affected by chronic exposure to sevoflurane. However, more randomized, controlled, and well-designed clinical studies with larger population are needed to confirm of these results.     

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.     

Epigallocatechin-3-gallate counters cisplatin toxicity of rat testes

Context: Epigallocatechin-3-gallate (EG), the main active flavonoid in green tea, has well-known anti-inflammatory, antioxidant, and anti-apoptotic activities.

Objective: The EG protection against testicular injury induced by cisplatin was studied in Sprague–Dawley rats.

Materials and methods: Cisplatin (10?mg/kg, i.p) was given as a single injection to rats. EG was given at 40 and 80?mg/kg/day, i.p., for 5 days, starting the same day of cisplatin insult. Serum testosterone, and testicular malondialdehyde, total antioxidant status, nitric oxide, interleukin-6, interleukin-1β, cytochrome C, Bax/Bcl-2 ratio, and caspase-3 were measured. In addition, testicular histopathological examination and immunohistochemical expression of testicular tumour necrosis factor-α were evaluated.

Results: Cisplatin, compared to the control, significantly decreased serum testosterone (6.48?±?0.7 vs. 50.8?±?4.91?ng/10?mL), and testicular tissue antioxidant status (17.3?±?1.21 vs. 64.12?±?5.4?μmol/g), and significantly increased interleukin-6 (85.81?±?6.11 vs. 38.2?±?2.79?pg/100?mg), interleukin-1β (98.09?±?8.31 vs. 32.52?±?2.08?pg/100?mg), malondialdehyde (74.5?±?5.88 vs. 23.8?±?1.91?nmol/g), nitric oxide (104.98?±?8.5 vs. 52.68?±?5.12?nmol/100?mg), cytochrome C (5.97?±?0.33 vs. 1.6?±?0.99?ng/mg protein), Bax/Bcl-2 ratio (4.01?±?0.38 vs. 0.71?±?0.0), and caspase-3 (3.2?±?0.21 vs. 0.98?±?0.08?O.D. 405?nm) in rat testes. EG (40 and 80?mg/kg, respectively) caused significant increases of serum testosterone (33.9?±?2.89 and 47.88?±?4.4?ng/10?mL), and testicular antioxidant status (47.1?±?3.92 and 58.22?±?3.58?μmol/g), and significant decreases of interleukin-6 (57.39?±?4.2 and 48.18?±?3.98?pg/100?mg), interleukin-1β (65.12?±?5.88 and 41.96?±?3.51?pg/100?mg), malondialdehyde (42.3?±?3.9 and 28.67?±?2.49?nmol/g), nitric oxide (70.6?±?6.79 and 61.31?±?5.18?nmol/100?mg), cytochrome C (3.4?±?0.27 and 2.21?±?0.18?ng/mg protein), Bax/Bcl-2 ratio (1.49?±?0.14 and 1.1?±?0.09), and caspase-3 (2.1?±?0.17 and 1.48?±?0.13?O.D. 405?nm) in testes of cisplatin-treated rats. Additionally, both doses of EG significantly ameliorated the histopathological injury and reduced tumour necrosis factor-α expression in rat testes.

Conclusion: EG can afford testicular protection in cisplatin-challenged rats by its antioxidant, antinitrative, anti-inflammatory and antiapoptotic effects.     

Rutin ameliorates oxidative stress and preserves hepatic and renal functions following exposure to cadmium and ethanol

Context: Rutin (RUT) is an antioxidant flavonoid with well-known metal chelating potentials.

Objective: This study was designed to evaluate the protective effects of RUT against cadmium (Cd)?+?ethanol (EtOH)-induced hepatic and renal toxicity in rats.

Materials and methods: Wistar rats were treated with Cd (50?mg/kg) alone or in combination with EtOH (5?mg/kg) and RUT (25, 50 and 100?mg/kg) for 15?days. After treatment, the liver, kidney and serum were removed for biochemical assays by spectrophotometric methods.

Results: Serum, hepatic and renal malondialdehyde (MDA) levels were highest in the Cd?+?EtOH group and lowest in Cd?+?EtOH animals co-treated with the highest dose of RUT (2.98?±?0.34, 10.08?±?2.32, 4.99?±?1.21 vs. 1.69?±?0.33, 6.13?±?0.28, 3.66?±?1.12?μmol MDA/mg protein, respectively). The serum level of Cd was increased in the Cd?+?EtOH treated animals compared to Cd?+?EtOH animals co-treated with 100?mg/kg RUT (2.54?±?0.08 vs. 1.28?±?0.04?ppm). Furthermore, RUT at the highest dose protected against Cd?+?EtOH-induced elevation of bilirubin and uric acid levels as well as activities of lactate dehydrogenase and γ-glutamyl transferase (62.86?±?2.74 vs. 122.52?±?6.35?µmol/L; 1.77?±?0.35 vs. 3.23?±?0.55?mmol/L; 9.56?±?1.22 vs. 16.21?±?1.64?U/L; 288.92?±?40.12 vs. 159.8?±?18.01?U/L). The histo-pathological changes in the liver and kidney were also reduced in the Cd?+?EtOH animals co-treated with RUT in a dose-dependent manner.

Discussion and conclusion: RUT protected against the combined effects of Cd?+?EtOH on hepatic and renal functions and improved the antioxidant defence system in the blood.     

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.     

Ferulic acid pretreatment mitigates MPTP-induced motor impairment and histopathological alterations in C57BL/6 mice

Abstract

Context: Ferulic acid (FA) is a potent ubiquitous plant antioxidant found in cereals such as brown rice, whole wheat, and oats. Phytochemical-based antioxidants are shown to be effective in neurodegenerative diseases. This study hypothesizes that supplementation of FA might combat oxidative stress-induced Parkinson’s disease (PD).

Objective: To explore the effect of FA on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP)-induced neurotoxicity.

Materials and methods: Mice were randomized into five groups: Group I mice served as control. Group II mice received 5?×?MPTP [25?mg/kg body weight (i.p.)] in saline 24?h apart starting from the 3rd day and continued till the last day of the experimental period of 7?d. In addition to MPTP injections, mice in Groups III, IV, and V were given FA at a dose of 20, 40, and 80?mg, respectively, for 7?d. Mice were subjected to a battery of behavioral tests along with histological investigations.

Results: Our histological findings revealed that MPTP administration enhanced Bax/Bcl2 ratio and microglial cells activation reflecting induction of apoptosis and inflammation, respectively. This dopaminergic neuronal loss caused impairment in motor balance and coordination in MPTP mice as assessed by various behavioral tests. FA at a dose of 40?mg/kg/d body weight effectively attenuated MPTP-induced neurotoxicity.

Discussion: Antioxidant, free-radical quenching, and anti-inflammatory activities of FA could contribute to its neuroprotective effect.

Conclusion: This study provides elementary evidence for the neuroprotective action of FA against MPTP-induced PD in mice and warrants further studies.     

Therapeutic role of quercetin on oxidative damage induced by acrylamide in rat brain

Context Quercetin (QE), a bioflavonoid present abundantly in fruits and vegetables, has been reported to possess antioxidant properties. Acrylamide (ACR) is formed in foods during cooking and is known to be neurotoxic.

Objective The present study was designed to evaluate the protective effect of QE against neurotoxicity induced by ACR.

Materials and methods Four groups of Wistar rats consisting of six rats each: (i) control group; (ii) acrylamide treated group (50?mg/kg body weight as single dose); (iii) quercetin group: rats were treated intraperitoneally (i.p.) with QE (10?mg/kg body weight alone every day for 5 d); (iv) quercetin?+?acrylamide group: quercetin (10?mg/kg bw) was given i.p. every day for 5 d followed by acrylamide i.p. injection (50?mg/kg bw) on fifth day (single dose). Rats were killed after 48?h.

Results Administration of ACR (50?mg/kg bw) in Wistar rats resulted in significant increase of dopamine, interferon-γ and 8-hydroxyguanosine with concomitant decrease of serotonin (p?<?0.001) in the rat brain. Treatment of rats with QE intraperitonealy (10?mg/kg body weight) before ACR assault resulted in the diminution of ACR-mediated neurotoxicity as evident from decreased levels of dopamine, interferon-γ (p?<?0.001) and 8-hydroxyguanosine with concomitant restoration of serotonin levels (p?<?0.001).

Discussion and conclusion On the basis of the above results, the present study suggests that quercetin may be a potential therapeutic agent for restoration of oxidative damage to neurons.     

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.     

Ameliorative effects of physcion 8-O-β-glucopyranoside isolated from Polygonum cuspidatum on learning and memory in dementia rats induced by Aβ1–40

Abstract

Context: Polygonum cuspidatum Sieb. Et Zucc. (Polygonaceae) has been traditionally used in folk medicine to treat various diseases.

Objective: This study investigates the ameliorative effects of physcion 8-O-β-glucopyranoside (PSG) isolated from P. cuspidatum on learning and memory in dementia rats induced by Aβ_1–40.

Materials and methods: Dementia rats were prepared by intracerebroventricular injection of Aβ_1–40. PSG (5, 10, 20, and 40?mg/kg/d, for 5?d) was administered orally. Ameliorative activity of PSG in dementia rats was evaluated by the Morris water maze (MWM) test, and its mechanisms were explored by evaluating AchE activity, levels of DA, NE, and 5-HT in hippocampus, and drebrin protein expressions in hippocampus.

Results: Our results indicated that PSG (5, 10, 20, and 40?mg/kg/d) significantly inhibited the prolonged latency in dementia rats (p?<?0.05), and inhibitory rates were 16.5, 22.7, 33.0, and 44.8% after 5?d of learning, indicating that PSG improves learning and memory of dementia rats. Furthermore, PSG significantly decreased AchE activity (10, 20, and 40?mg/kg/d; p?<?0.05), increased 5-HT (20 and 40?mg/kg/d, p?<?0.05), NE (10, 20, and 40?mg/kg/d; p?<?0.05), and DA levels (5, 10, 20, and 40?mg/kg; p?<?0.05) in the hippocampus. Additionally, PSG obviously decreased the Aβ contents in hippocampus (10, 20, and 40?mg/kg/d; p?<?0.05), and up-regulated drebrin protein expressions (5, 10, 20, and 40?mg/kg/d; p?<?0.05).

Conclusions: PSG can significantly enhance learning and memory in Aβ_1–40-induced dementia rats, and the mechanisms may be related to increase levels of Ach, 5-HT, NE, and DA, decrease Aβ contents, and up-regulation of drebrin proteins in hippocampus.     

Efficacy and safety of the single pill combination of amlodipine 10 mg plus valsartan 160 mg in hypertensive patients not controlled by amlodipine 10 mg plus olmesartan 20 mg in free combination

ABSTRACT

Background and objective: For patients with moderate hypertension (grade 2, defined as systolic blood pressure [SBP] 160–179?mmHg and/or diastolic blood pressure [DBP] 100–109?mmHg), current guidelines recommend initial combination therapy and rapid dose-adjustment to achieve blood pressure goal. In this study we investigated the efficacy and tolerability of the single pill combination of amlodipine 10?mg plus valsartan 160?mg (A?10?+?Val 160) in patients not controlled by the free combination of amlodipine 10?mg plus olmesartan 20?mg (A?10?+?O 20).

Methods: In this prospective, open-label, non-randomized trial, 257 patients with mean sitting DBP of 100–109?mmHg at trough entered a 4 week treatment phase with A?10?+?O 20 in free combination once daily. Patients in whom DBP remained uncontrolled were switched in a second 4 week treatment phase to A?10?+?Val 160. The primary efficacy variable was the reduction in DBP at week 8 compared to week 4 in the intent-to-treat population.

Results: In the total cohort, baseline SBP/DBP of 164.2?±?9.8/103.6?±?2.1?mmHg decreased by 19.2?±?12.4/14.1?±?7.4?mmHg at week 4. In patients who did not achieve BP control (n?=?175), subsequent treatment with A?10?+?Val 160 for 4 weeks reduced SBP from 149.6?±?11.1 at week 4 by 7.9?mmHg at week 8 (95% CI: 6.1–9.6, p?<?0.0001) and DBP from 93.4?±?3.9?mmHg by 9.1?mmHg (95% confidence interval: 8.1–10.2, p?<?0.0001). The combination of A?10?+?Val 160 was well tolerated, and the observed adverse events (15.3% of patients in phase 2) were consistent with the known drug profiles.

Conclusions: In a study designed to reflect typical clinical practice, in patients not controlled by the free combination of A?10?+?O 20, the single pill combination of A?10?+?Val 160 produced a statistically and clinically significant additional BP reduction and was well tolerated. Potential limitations of the design (open-label, non-controlled design, short term treatment) have to be taken into account.     

Effect of curcumin in mice model of vincristine-induced neuropathy

Abstract

Context: Curcumin exhibits a wide spectrum of biological activities which include neuroprotective, antinociceptive, anti-inflammatory, and antioxidant activity.

Objective: The present study evaluates the effect of curcumin in vincristine-induced neuropathy in a mice model.

Materials and methods: Vincristine sulfate (0.1?mg/kg, i.p. for 10 consecutive days) was administered to mice to induce neuropathy. Pain behavior was assessed at different days, i.e., 0, 7, 10, and 14?d. Sciatic nerve total calcium, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), nitric oxide (NO), and lipid peroxidation (LPO) were also estimated after the 14th day of study. Pregabalin (10?mg/kg, p.o.) and curcumin (15, 30, and 60?mg/kg, p.o.) were administered for 14 consecutive days.

Results: Curcumin at 60?mg/kg significantly attenuated the vincristine-induced neuropathic pain manifestations in terms of thermal hyperalgesia (p?<?0.001) and allodynia (p?<?0.001); mechanical hyperalgesia (p?<?0.001); functional loss (p?<?0.001); and in the delayed phase of formalin test (p?<?0.001). Curcumin at 30 and 60?mg/kg exhibited significant changes (p?<?0.001) in antioxidant levels and in total calcium levels in vincristine-injected mice.

Conclusion: Curcumin at 30 and 60?mg/kg dose levels significantly attenuated vincristine-induced neuropathy which may be due to its multiple actions including antinociceptive, calcium inhibitory, and antioxidant effect.     

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.     

The effects of Melissa officinalis (lemon balm) pretreatment on the resistance of the heart to myocardial injury

Context: The antihyperlipidemic, antiarrhythmic, neuroprotective and hepatoprotective effects of Melissa officinalis L. (Lamiaceae) have been reported. However, no study has examined its effects on the resistance of the heart to stressful conditions.

Objective: The objective of this study is to evaluate the effects of aqueous extract of M. officinalis aerial parts on Wistar rat heart with/without cardiac injury.

Materials and methods: Animals were grouped as control, isoproterenol (ISO), M. officinalis without (M50, M100, and M200) and with isoproterenol (M50?+?ISO, M100?+?ISO, and M200?+?ISO). The aqueous extract of M. officinalis was orally administered at dosages of 50, 100, and 200?mg/kg/d, respectively, for 7 consecutive days. On the 6th and 7th day, ISO, M50?+?ISO, M100?+?ISO, and M200?+?ISO groups received 85?mg/kg of isoproterenol for myocardial injury induction. On day 8, hemodynamic parameters were recorded and samplings were done.

Results: The extract (50, 100, and 200?mg/kg) significantly reduced the heart rate (264?±?5, 259?±?5 and 281?±?3 versus 377?±?13 in control group, p?<?0.01). Blood pressure was significantly decreased in M50?+?ISO (75?±?5) versus M50 (110?±?6) and M100?+?ISO (72?±?6) versus M100 (105?±?5?mmHg, p?<?0.01). The malondialdehyde levels of the injured hearts were lower in M50?+?ISO and M100?+?ISO groups than in the ISO group (p?<?0.05). Serum cardiac troponin I was higher in the M200?+?ISO group (5.1?±?1.7) than in the ISO group (2.7?±?0.7?ng/ml, p?<?0.05).

Conclusion: The lower dose of extract, by improving the balance of the redox system and by reducing the heart rate, may increase the heart resistance to injury. However, the higher doses of extract may intensify the injury of ischemic heart.     

Effect of pregabalin on fear-based conditioned avoidance learning and spatial learning in a mouse model of scopolamine-induced amnesia

Objectives: Cognitive deficits are one of the frequent symptoms accompanying epilepsy or its treatment.

Methods: In this study, the effect on cognition of intraperitoneally administered antiepileptic drug, pregabalin (10?mg/kg), was investigated in scopolamine-induced memory-impaired mice in the passive avoidance task and Morris water maze task. The effect of scopolamine and pregabalin on animals’ locomotor activity was also studied.

Results: In the retention phase of the passive avoidance task, pregabalin reversed memory deficits induced by scopolamine (p?p?p?p?Discussion: In passive avoidance task, pregabalin reversed learning deficits induced by scopolamine. In the Morris water maze, pregabalin did not influence spatial learning deficits induced by scopolamine. These results are relevant for epileptic patients treated with pregabalin and those who use it for other therapeutic indications (anxiety, pain).     

Assessment of luteolin isolated from Eclipta alba leaves in animal models of epilepsy

Context: Eclipta alba (Linn) Hassk. (Asteraceae) has been reported to be a nerve tonic and has been used to treat epilepsy in folk medicine.

Objective: The present study isolates and characterizes luteolin from E. alba and evaluates its antiepileptic potential in chemically induced acute and chronic models in mice.

Materials and methods: The methanol extract (16.85% w/w) of E. alba leaves was subjected to fractionation for isolation of luteolin. In acute pentylenetetrazole (PTZ) model, luteolin (5, 10, 20?mg/kg, i.p.) was administered 30?min prior to PTZ injection (100?mg/kg) in Swiss albino mice. Kindling was induced by chronic administration of PTZ (35?mg/kg) on every alternate day (48 days). Luteolin was investigated on the course of kindling development and oxidative stress markers [reduced glutathione (GSH) and malondialdehyde (MDA)] in kindled mice.

Results: Single-dose pretreatment with luteolin (10 and 20?mg/kg, i.p.) was found to be effective in an acute PTZ model (100% protection from mortality) and it did not exhibit any effect on motor coordination at the same doses. PTZ-induced kindling was significantly (p?p?p?Discussion and conclusion: The results of the present study demonstrated that luteolin had an anticonvulsant effect in an acute PTZ model. Luteolin exhibited and inhibitory effect on the course of kindling and associated oxidative stress and hence could be a potential molecule in the treatment of epilepsy.