Evaluation of Pistacia lentiscus seed oil and phenolic compounds for in vitro antiproliferative effects against BHK21 cells

Context: Within the global context of increasing cancer diseases, natural products are important in devising new drugs and providing unique ideas in cancer therapy. In Tunisian folk medicine, Pistacia lentiscus L. (Anacardiaceae) fixed oil is used for cancer treatment.

Objective: This investigation studied, for the first time, the antiproliferative effect of Pistacia lentiscus fixed oil and its phenolic extract on BHK21 cancer cells.

Materials and methods: Oil was extracted from fruits harvested in northwest Tunisia and the phenolic fraction was obtained by mixing with methanol. The anti-proliferative activity of the two tested substances on BHK 21 cells were investigated in vitro using trypan blue assays. Cells were treated with different concentrations of P. lentiscus oil (0.009, 0.018, 0.036, and 0.09 g/mL) and the phenolic extract (0.007, 0.014, 0.03, and 0.07 g/mL) for 24, 48, and 72 h.

Results: The inhibitory effect of Pistacia lentiscus fixed oil increases with the increase in dose. The IC₅₀ value was estimated at 0.029 g/mL. The percentage of cell viability was 42.46 ± 3.4% at a dose of 0.09 g/mL and was significantly lower than that of the untreated control (96.24 ± 2.5%, p<0.01). The phenolic extract demonstrated a dose- and time-dependent inhibitory effect on BHK21 cell growth. After 48 h of incubation, the IC₅₀ value was estimated at 0.15 g/mL.

Discussion and conclusion: The results demonstrated the potential of Pistacia lentiscus fixed oil in treating cancer, as it is used in traditional medicine.     

Anti-asthmatic property and possible mode of activity of an ethanol leaf extract of Polyscias fruticosa

Context: Polyscias fruticosa (L.) Harms (Araliaceae) is used as a traditional remedy for asthma in Ghana.

Objective: The objective of this study is to establish the anti-asthmatic property and a possible mode of activity of an ethanol leaf extract of P. fruticosa (PFE).

Materials and methods: The time (min) for pre-convulsive dyspnea, and time for recovery, after sensitization with 150?μg OVA and induction of bronchospasm with 1% acetylcholine or histamine in normal, and 100, 250, and 500?mg/kg PFE-treated Dunkin Hartley guinea pigs, were recorded. Atropine (0.1?mg), mepyramine (0.1?mg), and PFE (1?mg) effect on a contractile response of 2.0?×?10^?2?μg/ml acetylcholine and 5.8 × 10^?2?μg/ml histamine on the isolated guinea pig ileum was investigated. Cytological and histological studies were conducted using guinea pig peritoneal mast cells and mesenteric cells, respectively, to establish PFE effect on compound 48/80-induced mast cell degranulation.

Results: PFE (100–500?mg/kg) prolonged the onset of pre-convulsive dyspnea by 76.1–180.2% (p?≤?0.01–0.001), and decreased recovery time by 71.9–78.5% (p?≤?0.01–0.001). It also enhanced percentage protection against histamine-induced bronchospasm by 15.8–80.1-fold (p?≤?0.05–0.01), and decreased percentage recovery time 2.5–3.3-fold (p?≤?0.05–0.01). PFE significantly inhibited (60.4?±?8.3%) contractile responses of histamine and produced significant inhibition (56–79%: p?≤?0.001) of mast cell degranulation.

Conclusion: PFE has anti-asthmatic, antihistaminic, and mast cell stabilization effect making it useful in traditional asthma management.     

Antidepressant and anxiolytic activity of Lavandula officinalis aerial parts hydroalcoholic extract in scopolamine-treated rats

Context: Anxiety and depression are common in Alzheimer’s disease (AD). Despite some evidence, it is difficult to confirm Lavandula officinalis Chaix ex Vill (Lamiaceae) as an anxiolytic and antidepressant drug.

Objective: The effects of L. officinalis extract were studied in scopolamine-induced memory impairment, anxiety and depression-like behaviour.

Materials and methods: Male NMRI rats were divided into control, scopolamine alone-treated group received scopolamine (0.1?mg/kg) intraperitoneally (i.p.), daily and 30?min prior to performing behavioural testing on test day, for 12 continuous days and extract pretreated groups received aerial parts hydro alcoholic extract (i.p.) (100, 200 and 400?mg/kg), 30?min before each scopolamine injection. Memory impairment was assessed by Y-maze task, while, elevated plus maze and forced swimming test were used to measure anxiolytic and antidepressive-like activity.

Results: Spontaneous alternation percentage in Y maze is reduced by scopolamine (36.42?±?2.60) (p?≤?0.001), whereas lavender (200 and 400?mg/kg) enhanced it (83.12?±?5.20 and 95?±?11.08, respectively) (p?≤?0.05). Also, lavender pretreatment in 200 and 400?mg/kg enhanced time spent on the open arms (15.4?±?3.37 and 32.1?±?3.46, respectively) (p?≤?0.001). On the contrary, while immobility time was enhanced by scopolamine (296?±?4.70), 100, 200 and 400?mg/kg lavender reduced it (193.88?±?22.42, 73.3?±?8.25 and 35.2?±?4.22, respectively) in a dose-dependent manner (p?≤?0.001).

Discussion and conclusion: Lavender extracts improved scopolamine-induced memory impairment and also reduced anxiety and depression-like behaviour in a dose-dependent manner.     

Risk assessment of low arsenic exposure using biomarkers of oxidative and genotoxic stress in a piscine model

The high level exposure to arsenic induces marked oxidative and genotoxic stress. However, information on the potential of low level arsenic exposure in this context is still scanty. In the present study, the extent of oxidative stress and genetic toxicity induced by low arsenic exposure was explored in freshwater fish Channa punctatus. Fish were exposed to low levels of arsenic (10 and 50 µg L⁻¹) as well as to its high level (500 µg L⁻¹) using sodium arsenite in aquaria water for 14 consecutive days. The TBARS assay for lipid peroxidation exhibited the increased occurrence of oxidative damage in the erythrocytes of fish at both the lower and higher levels of arsenic exposure. The level of reduced glutathione was also elevated in all the three arsenic exposed groups of fish compared to control. In contrast, significant decline was observed in the levels of three major antioxidant enzymes namely, superoxide dismutase, catalase and glutathione peroxidase, upon exposure to higher as well as lower levels of arsenic. Significant increases in micronucleus induction were found in the erythrocytes of fish even at the low levels of arsenic exposure. The study further revealed the occurrence of DNA fragmentation in the erythrocytes of fish at low arsenic exposures as well. The low level exposure to arsenic (using sodium arsenite), therefore, appeared to be capable of inducing noticeable oxidative stress as well as potential genotoxic effect in Channa punctatus. Moreover, the ability of arsenic to induce oxidative stress invariably appeared correlated with its genotoxic potential.

     

Modulation of cellular antioxidant defense activities by sodium arsenite in human fibroblasts

 Many studies have shown that oxygen radicals can be produced during arsenic metabolism. We report here that in human fibroblasts (HFW cells) sodium arsenite exposure caused increased formation of fluorescent dichlorofluorescein (DCF) by oxidation of the nonfluorescent form. The enhanced DCF fluorescence was inhibited by a radical scavenger, butylated hydroxytoluene. The effects of sodium arsenite treatment on cellular antioxidant activities were then examined. Treatment of HFW cells with sodium arsenite resulted in a significant increase in heme oxygenase activity and ferritin level. Sodium arsenite-enhanced heme oxygenase synthesis was inhibited by co-treatment of cells with the antioxidants sodium azide and dimethyl sulfoxide. Furthermore, sodium arsenite treatment did not apparently affect glucose-6-phosphate dehydrogenase activity, but resulted in significantly increased glutathione levels and superoxide dismutase activity, slightly decreased glutathione peroxidase activity, and significantly decreased catalase activity. Sodium arsenite toxicity was partly reduced by addition of catalase to the culture medium. These results imply that arsenite can enhance oxidative stress in HFW cells. Received: 24 November 1994 / Accepted: 31 January 1995     

Prevention of arsenic-mediated reproductive toxicity in adult female rats by high protein diet

Abstract

Context: The detrimental effects of arsenic on female reproductive functions may involve overt oxidative stress. Casein and pea [Pisum sativum Linn. (Fabaceae)] proteins have antioxidant properties.

Objective: To investigate the role of casein- and pea-supplemented high-protein diet (HPD) in utero-ovarian protection from arsenic toxicity.

Materials and methods: Adult female Wistar rats were orally gavaged with vehicle (Gr-I) or arsenic at 3?ppm/rat/d (Gr-II and Gr-III) for 30 consecutive days, when they were maintained on either regular diet containing 18% protein (Gr-I and Gr-II), or HPD containing 27% protein in the form of casein (20%) and pea (7%) (Gr-III). Reproductive functions were evaluated using a battery of biochemical and histological techniques.

Results: As compared to Gr-I, the Gr-II rats suffered from loss of estrous cyclicity, reduction in weight (mg/100?g body weight) of ovary (Gr-I: 54.3?±?4.2 versus Gr-II: 35.8?±?1.6; p?<?0.001) and uterus (Gr-I: 161.7?±?24.6 versus Gr-II: 94.44?±?13.2; p?<?0.05), utero-ovarian degeneration, attenuated ovarian activities (unit/mg tissue/h) of Δ⁵, 3β-hydroxysteroid dehydrogenase (Gr-I: 3.41?±?0.12 versus Gr-II: 2.31?±?0.09; p?<?0.01) and 17β-hydroxysteroid dehydrogenase (Gr-I: 3.82?±?0.57 versus Gr-II: 1.24?±?0.19; p?<?0.001), and decreased serum estradiol level (pg/ml) (Gr-I: 61.5?±?2.06 versus 34.1?±?2.34; p?<?0.001). Ovarian DNA damage was preponderant with blatant generation of malondialdehyde (nM/mg tissue; Gr-I: 15.10?±?2.45 versus Gr-II: 29.51?±?3.44; p?<?0.01) and attenuated superoxide dismutase activity (unit/mg tissue) (Gr-I: 2.18?±?0.19 versus Gr-II: 1.33?±?0.18; p?<?0.05). The Gr-III rats were significantly protected from these ill effects of arsenic.

Discussion and conclusion: HPD, by way of antioxidant properties, may find prospective role in the protection of reproductive damage caused by arsenic.     

Chemical Composition of Essential Oils from Leaves and Twigs of Pistacia lentiscus,Pistacia lentiscus var. chia,and Pistacia terebinthus from Turkey

The composition of the essential oils from the leaves and twigs of Pistacia lentiscus L., Pistacia lentiscus var. chia (L.), and Pistacia terebinthus L. of Turkish origin were analyzed by gas chromatography/mass spectrometry (GC/MS). Seventy-seven constituents were characterized from the essential oil of P. terebinthus leaves with α-cadinol (6.9%), phytol (5.4%), δ-cadinene (5.1%), α-terpineol (5.0%), and bornyl acetate (4.4%) as major constituents. Germacrene D (10%), β-pinene (7.5%), bornyl acetate (6.0%), α-cubebene (5.9%), and cubebol (5.4%) were found to be the main components among the 61 compounds characterized in the essential oil of P. terebinthus twigs. Forty-six compounds were characterized from the essential oil of P. lentiscus twigs with sabinene (23.2%), α-pinene (19.4%), germacrene D (14.1%), limonene (6.9%), β-phellandrene (6.5%), terpinene-4-ol (5.7%), and β-caryophyllene (5.7%) as the main constituents. Terpinene-4-ol (29.2%), β-caryophyllene (29.2%), and p-cymene (7.1%) were identified as the major components among the 64 compounds characterized in the essential oil of P. lentiscus leaves. Sixty-eight compounds were found on the essential oil of P. lentiscus var. chia leaves with germacrene D (20.1%), myrcene (13.9%), β-caryophyllene (10.8%), and α-terpinyl acetate (4.8%) as the major constituents. Myrcene (27.4%), germacrene D (21.7%), and β-caryophyllene (7.2%) were found to be the main components among 50 compounds characterized in the essential oil of P. lentiscus var. chia twigs.     

Mitigating role of quercetin against sodium fluoride-induced oxidative stress in the rat brain

Context: Quercetin is a well known aglycone flavonoid that is widely found in different food sources.

Objective: In this study, the in vivo neuroprotective potential of quercetin against sodium fluoride-induced oxidative stress was evaluated.

Materials and methods: Wistar rats were divided into five treatment groups and then subjected to daily intraperitoneally treatment with quercetin (at 10 and 20?mg/kg body weight), vitamin C (at 10?mg/kg), or vehicle. After a 1 week treatment period, all groups except saline treated (normal group), were intoxicated with sodium fluoride (NaF) for 1 week. Rat brains were then removed and homogenized for measurement of antioxidant markers including superoxide dismutase (SOD), reduced glutathione, catalase, and lipid peroxidation final products.

Results: The thiobarbituric acid reactive substances (TBARS) levels in the heart homogenate of sodium fluoride treated rats (42.04?±?2.14 nmol MDA eq/g tissue) increased compared to the normal rats (35.99?±?1.08 nmol MDA eq/g tissue). Animals which were pretreated with quercetin at 20?mg/kg for 1 week prior to sodium fluoride intoxication showed significant reduction in the TBARS level (36.13?±?1.12 nmol MDA eq/g tissue). Also, pretreatment with quercetin (20?mg/kg) restored the SOD and catalase activities and modified the level of reduced glutathione compared with the control group (p?>?0.05).

Discussion and conclusion: The present study revealed a potent neuroprotective potential of quercetin against NaF-induced toxicity.     

Protection of arsenic-induced hepatic disorder by arjunolic acid

Arsenic is one of the ubiquitous environmental pollutants, which affects nearly all organ systems. The present study has been carried out to investigate the hepatoprotective role of arjunolic acid, a triterpenoid saponin, against arsenic-induced oxidative damages in murine livers. Administration of sodium arsenite at a dose of 10 mg/kg body weight for 2 days significantly reduced the activities of antioxidant enzymes, superoxide dismutase, catalase, glutathione S-transferase, glutathione reductase and glutathione peroxidase as well as depleted the level of reduced glutathione and total thiols. In addition, sodium arsenite also increased the activities of serum marker enzymes, alanine transaminase and alkaline phosphatase, enhanced DNA fragmentation, protein carbonyl content, lipid peroxidation end-products and the level of oxidized glutathione. Studies with arjunolic acid show that in vitro it possesses free radical-scavenging and in vivo antioxidant activities. Treatment with arjunolic acid at a dose of 20 mg/kg body weight for 4 days prior to arsenic administration prevents the alterations of the activities of all antioxidant indices and levels of the other parameters studied. Histological studies revealed less centrilobular necrosis in the liver treated with arjunolic acid prior to arsenic intoxication compared to the liver treated with the toxin alone. Effects of a known antioxidant, vitamin C, have been included in the study as a positive control. In conclusion, the results suggest that arjunolic acid possesses the ability to attenuate arsenic-induced oxidative stress in murine liver probably via its antioxidant activity.     

Oxidative stress status in patients with acute urticaria

Background: The conflicting information related to oxidative stress status in patients with chronic idiopathic urticaria has been reported in several studies. However, the association between acute urticaria (AU) and oxidative stress has not been investigated exhaustively.

Objectives: To evaluate the role of the oxidative stress in the patients with AU by determining the oxidant/antioxidant activity in AU and to establish its clinical significance.

Methods: About 50 patients with AU, (10 males, 40 females) and 30 unrelated healthy controls (4 males, 26 females) were enrolled into the study. The activity of the antioxidant enzymes copper-zinc superoxide dismutase (Cu-Zn-SOD), glutathione peroxidase (GSH-Px) and catalase, and the levels of malondialdehyde (MDA), serum NO and protein carbonyls levels in the plasma were measured spectrophotometrically at samples.

Results: A statistically significant increase was observed in serum Cu-ZnSOD activities of the patients when compared with that of the controls (p?p?=?0.002). Serum MDA and NO levels were significantly higher in patients with AU when compared with control group (p?p?Conclusion: It seems there is an oxidative burden in the patients with AU. Cutaneous oxidative stress may play a role in pathogenesis of the disease.     

Protective effect of Corchorus olitorius leaves on sodium arsenite-induced toxicity in experimental rats

The present study was undertaken to evaluate the protective effect of aqueous extract of Corchorus olitorius leaves (AECO) against sodium arsenite-induced toxicity in experimental rats. The animals exposed to sodium arsenite at a dose of 10 mg/kg body weight p.o. for 10 days exhibited a significant inhibition (p < 0.01) of hepatic and renal antioxidant enzymes namely superoxide dismutase, catalase, glutathione-S-transferase, glutathione peroxidase and glutathione reductase. In addition, arsenic intoxication significantly decreased (p < 0.01) the level of reduced glutathione and increased (p < 0.01) the levels of oxidized glutathione and thiobarbituric acid reactive substances in selected tissues. Treatment with AECO at doses of 50 and 100 mg/kg body weight p.o. for 15 days prior to arsenic intoxication significantly improved hepatic and renal antioxidant markers in a dose dependant manner. AECO treatment also significantly reduced the arsenic-induced DNA fragmentation of hepatic and renal tissues. Histological studies on the ultrastructural changes of liver and kidney supported the protective activity of the AECO. The results concluded that the treatment with AECO prior to arsenic intoxication has significant role in protecting animals from arsenic-induced hepatic and renal toxicity.     

Induction of anoikis by sodium arsenite in rat hepatoma FGC4 cells: comparison with cadmium chloride and implications for assessment of regulation of heat shock protein 70

Context: Arsenic, a toxic metalloid with major health concerns, elicits upregulation of heat shock protein 70 (HSP70) in rat hepatoma FGC4 cells, together with evidence of detachment of viable cells from the growth substratum.

Objective: To determine if this cell detachment was linked to anoikis, and the impact of this on measurement of HSP70 expression.

Materials and methods: FGC4 cells were exposed to sodium arsenite, and detached and attached cells were taken for assessment of cell viability, activation of procaspase-3, and expression of HSP70.

Results: Exposure to sodium arsenite led to loss of viable cells from the substratum, associated with apoptosis in detached, but not attached, cells. Upregulation of HSP70 of a similar magnitude was demonstrated in both cell populations. Exposure of cells to cadmium chloride, a toxic metal, also of major environmental concern and believed to act by an oxidative stress mechanism, produced very little release of viable cells from the culture substratum, was not associated with apoptosis, but did elicit a modest upregulation of HSP70 in both cell populations.

Discussion: Exposure of FGC4 cells to sodium arsenite elicits anoikis, a form of anchorage-dependent apoptosis, and assessment of the level of HSP70 upregulation in such cells should take account of the detached cell population. Further, the data suggest that this phenomenon is selective to sodium arsenite, rather than to another toxic element that shares a similar mechanism of toxicity.     

Effects of Pycnogenol on cisplatin-induced optic nerve injury: an experimental study

Aim: To determine the effects of Pycnogenol on cisplatin-induced optic nerve damage.

Material and method: Totally 18 albino Wistar male rats were assigned into three groups, with six rats in each group as follows: healthy controls (HC group), only cisplatin (2.5?mg/kg) administered group (CIS group) and Pycnogenol (40?mg/kg)?+?cisplatin (2.5?mg/kg) administered group (PYC group). We analyzed the levels of malondialdehyde (MDA) as a marker of lipid peroxidation and oxidative stress, total glutathione (tGSH) as a marker of antioxidant status, nuclear factor-kappa B (NF-κB) and tumor necrosis factor alpha (TNF-α) as inflammatory markers, total oxidative status (TOS) and total antioxidant status (TAS) on eye tissue together with histopathological evaluation of optic nerve in an experimental model.

Results: In CIS group MDA, TOS, TNF-α and NF-κB levels were statistically significantly higher (p?p?p?p?Conclusion: Pycnogenol pretreatment was highly effective in preventing augmentation of cisplatin-induced oxidative stress and inflammation in eye tissue.     

Antioxidant effects of piperine in in-vivo chick embryo cataract model induced by steroids

Purpose: The etiopathogenesis of steroid-induced cataracts is unknown. One hypothesis is that the higher reactive oxygen species (ROS) levels play an important role in the pathogenesis of several disorders, including the evolution of cataracts. This study investigated the antioxidant effects of piperine in our steroid-induced chick embryo lens model.

Methods: The study included 36 specific pathogen-free (SPF) fertilized eggs divided into six groups: phosphate buffer saline (PBS, pH 7.4 Saline Solution (0.9%) isotonic) group (G1), hydrocortisone succinate sodium (HC)-treated group (G2), 100?mg/kg piperine and HC treated group (G3), 50?mg/kg piperine and HC treated group (G4), 25?mg/kg piperine and HC treated group (G5), and 10?mg/kg piperine and HC treated group (G6). On the 15th day of incubation, the SPF eggs in the six groups were removed from the incubator; all were injected using insulin injectors into the chorioallantoic membrane. On day 17, all of the chick embryos were removed from the eggs and all lenses were dissected from the embryos. Cataract formation was evaluated in all lenses, and total antioxidant status (TAS), total oxidant status (TOS), reduced glutathione (GSH), and lipid peroxidation (MDA, malondialdehyde) levels were measured in all lens.

Results: The lenses in the G1 group had higher levels of GSH and TAS (p?p?p?p?p?p?p?Conclusion: Steroid therapy causes a decrease in GSH and TAS levels and an increase in TOS and MDA levels in lens tissues, indicating increased oxidative stress. Piperine exerts its effects as an antioxidant substance, in increasing doses.     

Effect of Arsenic (V) on the Antioxidant Defense System: In vitro Oxidation of Rat Plasma Lipoprotein

Abstract: Adult male rats were treated orally with sodium arsenate (10 mg As/ kg/day) for 2 days, and an increase in hepatic glutathione level was seen. Ascorbic acid content increased in both liver and plasma of intoxicated animals. Hepatic activities of superoxide dismutase and glutathione peroxidase did not change with the treatment and there was no increase in the level of lipid peroxidation measured as thiobarbituric acid-reacting substances (TBARS). Arsenic decreased the plasma level of uric acid and increased the plasma triglycerides content without modifying vitamin E levels. Both total lipoproteins and very low density lipoprotein plus low density lipoprotein (VLDL+LDL) fractions demonstrated greater propensity for in vitro oxidation than the corresponding untreated rats. The last finding might be a useful parameter for determining the degree of oxidative stress in the initial steps of intoxication with arsenic.     

Effect of fruit vinegars on liver damage and oxidative stress in high-fat-fed rats

Context: Vinegar has long been used as a condiment and a traditional medicine worldwide.

Objective: The current study investigates the antioxidant effect of three types of fruit vinegars (FV) namely pomegranate [Punica granatum L. (Punicaceae)], prickly pear [Opuntia ficus-indica (L.) Mill. (Cactaceae)], and apple [Malus domestica Borkh. (Rosaceae)] vinegars in high-fat diet (HFD)-induced hyperlipidemic Wistar rats.

Materials and methods: Fifty male Wistar rats were divided into five groups; HFD (80?cal/d) fed rats were orally dosed with fruit vinegars (7?ml/kg) once daily for 28?weeks. At the end of the experiment, lipid profile, lipid peroxidation products, antioxidant enzymes, and trace elements were assessed in serum. In addition, a liver histopathological study was performed.

Results: HFD showed a significant increase (p?≤?0.05) in lipid profile and TBARS levels when compared with normal control. Daily oral administration of FV normalized various biochemical, metabolic, and histopathological changes. However, pomegranate vinegar exhibited a very significant (p?≤?0.001) reduction in lipid profile levels (total cholesterol: 165%, triglycerides: 68%, LDL-c: 76%, and atherogenic index: 80%), whereas an increase in antioxidant status (SOD: 7-fold, GPx: 4.81-fold, GRx: 1.66-fold, and TAS: 3.45-fold) when compared with hyperlipidemic control. Histopathological examinations also confirmed the protective effects of pomegranate vinegar against lipid accumulation and the improvement of hepatic lesions.

Discussion and conclusion: The fruit vinegars regulate lipid metabolism and decrease liver damage in high-fat fed rats as shown in this study.     

Effect of triptolide on reproduction of female lesser bandicoot rat,Bandicota bengalensis

Abstract

Triptolide has been reported to cause antifertility in male rats and mice. However, studies on female rats have been limited. Present study was aimed to evaluate the effects of triptolide on reproduction of wild female rodent pest species, Bandicota bengalensis. Feeding of bait containing 0.1, 0.15 and 0.2% triptolide for a period of 15 days in bi-choice resulted in per day ingestion of 17.37, 23.54 and 27.49?mg/kg body weight of triptolide, respectively. Examination of vaginal smear of all the rats revealed a significant (p?≤?0.05) increase in duration of estrous cycle due to increase in durations of metestrous and diestrous stages in rats of treated groups. Autopsy of rats after 15 and 30 days of treatment withdrawal revealed significant (p?≤?0.05) reduction in weights of uterus and ovaries, non-significant reduction in weights of liver and levels of estradiol and progesterone and significant (p?≤?0.05) reduction in levels of urea and BUN and increase in levels of plasma proteins, ALT, AST, ALP, ACP and LDH in rats of treated groups compared to untreated group. There was no significant (p?≤?0.05) effect of treatment on body weight. Triptolide treatment affected the histomorphology of uterus by causing a decrease in lumen and columnar cell height and number of uterine glands and ovary by increasing the number of atretic follicles and decreasing the number of developing follicles. The present study suggests triptolide to be a strong candidate affecting reproduction of female B. bengalensis.     

Alpha-lipoic acid protects oxidative stress,changes in cholinergic system and tissue histopathology during co-exposure to arsenic-dichlorvos in rats

We investigated protective efficacy of α-lipoic acid (LA), an antioxidant against arsenic and DDVP co-exposed rats. Biochemical variables suggestive of oxidative stress, neurological dysfunction, and tissue histopathological alterations were determined. Male rats were exposed either to 50 ppm sodium arsenite in drinking water or in combination with DDVP (4 mg/kg, subcutaneously) for 10 weeks. α-Lipoic acid (50 mg/kg, pos) was also co-administered in above groups. Arsenic exposure led to significant oxidative stress along, hepatotoxicity, hematotoxicity and altered brain biogenic amines levels accompanied by increased arsenic accumulation in blood and tissues. These altered biochemical variables were supported by histopathological examinations leading to oxidative stress and cell death. These biochemical alterations were significantly restored by co-administration of α-lipoic acid with arsenic and DDVP alone and concomitantly. The results indicate that arsenic and DDVP induced oxidative stress and cholinergic dysfunction can be significantly protected by the supplementation of α-lipoic acid.     

3,3′-Diindolylmethane enhances apoptosis in docetaxel-treated breast cancer cells by generation of reactive oxygen species

Context: A major problem in the treatment of cancer is the development of toxic side effects and resistance to chemotherapy. The use of plant compounds to overcome resistance and prevent toxicity is a potential strategy for treatment.

Objective: We evaluated whether 3,3′-diindolylmethane (DIM) enhanced the sensitivity of breast cancer cells to docetaxel (DOC).

Materials and methods: MDA-MB231 and Sk-BR-3 cells were treated with and without 25 or 50?µM of DIM and 1?nM of DOC for 48 and 72?h, respectively. MTT assay was used to measure cell survival. Apoptosis and intracellular reactive oxygen species (ROS) were determined by flow cytometry. The expression of proteins regulating ROS production and apoptosis was evaluated by immunoblotting technique.

Results: Combining 25?µM of DIM with 1?nM DOC decreased cell survival by 42% in MDA-MB231 cells and 59% in Sk-BR-3 cells compared to control, DIM, or DOC (p?≤?0.05). The combination treatment increased apoptosis over 20% (p?≤?0.01) in both cell lines, which was associated with decreased Bcl-2, increased Bax, cleaved PARP and activated JNK (p?≤?0.01). ROS production increased by 46.5% in the MDA-MB231 and 29.3% in Sk-BR-3 cells with the combination compared to DIM or DOC alone. Pretreating cells with N-acetyl-cysteine or Tiron abrogated the anti-survival effect of the combination. The increase in ROS was associated with a 54% decrease in MnSOD and 47% increase in NOX2 protein compared to the other groups.

Conclusions: Our findings indicated that DIM enhances the sensitivity of breast cancer cells to DOC treatment by increasing ROS, which led to decreased cell survival and apoptosis.     

Efficacy and safety of the dipeptidyl peptidase-4 inhibitor alogliptin added to pioglitazone in patients with type 2 diabetes: a randomized,double-blind,placebo-controlled study

ABSTRACT

Objectives: To evaluate the efficacy and safety of alogliptin in patients with type 2 diabetes inadequately controlled by therapy with a thiazolidinedione (TZD).

Research design and methods: In a multicenter, double-blind, placebo-controlled clinical study, 493 patients 18–80 years old with inadequate glycemic control after stabilization (i.e., glycosylated hemoglobin [HbA_1c] 7.0–10.0%) despite ongoing treatment with a TZD were randomly assigned (2:2:1) to treatment with pioglitazone plus alogliptin 12.5?mg, alogliptin 25?mg or placebo once daily. Concomitant therapy with metformin or sulfonylurea at prestudy doses was permitted.

Main outcome measures: The primary efficacy endpoint was change in HbA_1c from baseline to Week 26. Secondary endpoints included changes in fasting plasma glucose (FPG) and body weight, and incidences of marked hyperglycemia (FPG?≥?200?mg/dL [11.10?mmol/L]) and rescue for hyperglycemia.

Results: Least squares (LS) mean change in HbA_1c was significantly (p?<?0.001) greater for alogliptin 12.5?mg (?0.66%) or 25?mg (?0.80%) than for placebo (?0.19%). A significantly (p?≤?0.016) larger proportion of patients achieved HbA_1c?≤?7% with alogliptin 12.5?mg (44.2%) or 25?mg (49.2%) than with placebo (34.0%). LS mean decreases in FPG were significantly (p?=?0.003) greater with alogliptin 12.5?mg (?19.7?mg/dL [?1.09?mmol/L]) or 25?mg (?19.9?mg/dL [?1.10?mmol/L]) than with placebo (?5.7?mg/dL [?0.32?mmol/L]). The percentage of patients with marked hyperglycemia was significantly (p?<?0.001) lower for alogliptin (≤25.0%) than placebo (44.3%). The incidences of overall adverse events and hypoglycemia were similar across treatment groups, but cardiac events occurred more often with active treatment than placebo.

Conclusions: Addition of alogliptin to pioglitazone therapy significantly improved glycemic control in patients with type 2 diabetes and was generally well tolerated. The study did not evaluate the effect of combination therapy on long-term clinical outcomes and safety.

Clinical trial registration: NCT00286494, clinicaltrials.gov.