Study of antidiabetic activity of two novel Schiff base derived dibromo and dichloro substituted compounds in streptozotocin-nicotinamide-induced diabetic rats: pilot study

Schiff bases are aldehyde-or ketone-like chemical compounds in which an imine or azomethine group replaces the carbonyl group. Such compounds show various beneficial biological activities, such as anti-inflammation and antioxidants. The present study addresses comprehensiveevaluation of antidiabetic effect of two novel dibromides and dichlorides substituted Schiff bases substituted Schiff bases (2,2'-[1,2-cyclohexanediylbis (nitriloethylidyne)]bis[4-chlorophenol] (CNCP) and 2, 2'-[1,2-cyclohexanediylbis(nitriloethylidyne)]bis[4-bromophenol] (CNBP) with two different doses, high (LD) and low (LD) in streptozotocin and nicotinamide induced diabetic rats. The rats were separated into normal, untreated, treated and reference groups. Except for the normal group, diabetes traits were induced in the rest animals. Insulin level was measured, and the effect of the compounds on biochemical parameters of liver function and lipid profile were evaluated. High glucose and decreased insulin level are observed in the groups. The histological evaluation confirms that the hepatic architecture in the treated animals with a low dose of CNCP is quite similar to that of the normal hepatic structure and characterized by normal central vein, hepatocytes without any fatty alterations and mild red blood cell infiltration. CNCP (LD) and CNBP (HD) are more successful in enhancing cell survival in the diabetic rat’s liver and can be responsible for causing much healthier structure and notable morphology improvement

The effect of cinnamomum cassia on two breast cancer cell lines / Sima Kianpour Rad

The bark of the cinnamon tree (Cinnamomum cassia) is a popular culinary spice. It is also used in traditional medicine to maintain health and prevent disease. The antioxidant and anticancer activity of C. cassia was investigated using various assays. C. cassia bark was sequentially extracted with seven solvents of varying polarity. The acetone extract of C. cassia, at 30 μg/ml, protected the mouse fibroblast cell line, 3T3-L1, from DNA damage by 45 %, as estimated by the comet assay. The acetone extract had the highest total phenolic and flavonoid content. The hexane extract of C. cassia and the two main components, trans-cinnamaldehyde and coumarin, inhibited the proliferation of two breast cancer cell lines, the estrogen-sensitive MCF-7 cells (IC50, 34 ±3.52 μg/ml) and the estrogen-insensitive MDA-MB-231 cells (IC50, 32.42 ±0.37 μg/ml). The mechanism of cell death was investigated by determining the activity of the caspases. The expression of particular apoptotic genes such as Bcl2, Akt1, p53 and Bid were investigated by real time RT-PCR. The hexane extract activated initiator caspases-8 and -9 and effector caspases-3 and -7. There was up-regulation of Bid and p53 expression. Akt1 expression was down-regulated in MDA-MB-231cells but up-regulated in MCF-7 cells, indicating partial resistance to apoptosis. The activity of the antioxidant enzymes, catalase and glutathione peroxidase in both cell lines, in response to 100 μg/ml of hexane extract, decreased in a time dependent manner, whereas that of superoxide dismutase decreased in MDA-MB-231 cells but increased in MCF-7 cells, indicating that C. cassia bark is a good source of antioxidants. Together with its anticancer and anticarcinogenic properties, it is a good supplement for maintenance of health and prevention of cancer

Anti-pain and anti-inflammation like effects of Neptune krill oil and fish oil against carrageenan induced inflammation in mice models: Current statues and pilot study

Although inflammation is a reactive to injurious stimuli and considered as beneficial process in body, but it causes some discomforts, such as pain. Murine dietary contains appreciable amounts of fatty acids and antioxidants which encourages researchers to focus on their potential therapeutic effects. This study is aimed to examine the analgesic and anti-inflammatory activity of Neptune krill oil (NKO) and fish oil (FO) in rodent model which are two well-known sources of rich content of n-3 polyunsaturated fatty acids (n-3 PUFAs), mostly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). NKO and FO were used at the same dose of 500 mg and also balanced at similar doses of EPA: 12 in NKO vs. 12 in FO wt%, DHA: 7 NKO vs. 8 FO wt%. Application of NKO and FO in acetic acid-induced writhing effect, hot plate, and formalin induced test, indicated the nociceptive activity of the two tested drugs in comparison with normal saline. Also, the anti-inflammatory effect of these supplements was confirmed by carrageenan test. Analysis of cytokines levels in the blood samples of the mice after induction inflammation by carrageenan indicated decreased levels of those proteins compared to that in the normal groups. Both tested drugs, effectively could reduce severe inflammation and pain in rodents in comparison with the references drugs (depends on the tests); however, NKO was found to be more effective. Keywords: Anti-nociceptive, Anti-inflammation, Neptune krill oil, Fish oil, n-3 PUFAs, EPA, DHA, IL-6, TNF-

In vivo evaluation of wound healing improvement of a new Schiff base derived bromine compound (CNBP) in rats

Background: The effect of delivered bromine vapor and also bromine substitutions are shown to play an important role in anti-inflammatory activity. The present study encompasses a broad in vivo study to size up healing activity of a novel dibromide substituted Schiff based compound against cutaneous wound model in Sprague Dawley (SD) rats. Methodology/principal findings: 2, 2′-[1, 2-Cyclohexanediylbis(nitriloethylidyne)] bis[4-bromophenol], CNBP, is synthesized through a Schiff base reaction applying the related ketone and diamine as the initiators. Four groups of six in each male SD rats are divided as negative group which are treated with gum acacia, positive control animals which are treated with topical dosage of Intrasite gel, and testing groups treated with low (10 mg/kg) and high (20 mg/kg) doses of CNBP. The excisional wounds are created on the posterior neck area of each group and the wound closure percentage was measured in the two separated days of the experiment. Moreover, the histopathological evaluation and determination of activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (GPx), and malondialdehyde (MDA) of the skin sections are performed. Furthermore, the immunohistochemistry consists of the evaluation of Hsp70 and BAX proteins. Conclusions/significance: The wound closure percentage showed a significant increase in high dose CNBP-treated group compared to the negative control. Histopathological evaluation of the skin sections showed that granulation tissue contained more proliferating fibroblast, collagen deposition, angiogenesis, and also less inflammatory cells in the high dose CNBP-treated group compared to the normal rats. In the treated groups with CNBP, SOD, CAT, and GPx activities were found significantly higher, however, the MDA level was shown to be lower (*P < 0.05; **P < 0.01; ***P < 0.001) than the negative control. At the molecular level, CNBP (20 mg/ml, HD) improved wound-healing process via down and up regulation of Bax and Hsp70 protein, respectively at the wound site. © 201

Gastroprotective activity of a novel Schiff base derived dibromo substituted compound against ethanol-induced acute gastric lesions in rats

Abstract Background Basic function of bromine in body is to activate pepsin production in gastritis with low acidity. The present study encompasses a broad in vivo study to evaluate gastroprotective activity of a novel dibromo substituted Schiff base complex against Sprague Dawley (SD) rats. Methods 2, 2′-[1, 2-cyclohexanediylbis (nitriloethylidyne)]bis(4-bromophenol) (CNBP) is synthesized via a Schiff base reaction, using the related ketone and diamine as the starting materials. SD rats are divided as normal, ulcer control (5 ml/kg of 10% Tween 20), testing (10 and 20 mg/kg of CNBP) and reference groups (omeprazole 20 mg/kg). Except for the normal group, the rest of the groups are induced gastric ulcer by ethanol 1 h after the pre-treatment. Ulcer area, gastric wall mucus, and acidity of gastric content of the animal stomachs are measured after euthanization. Antioxidant activity of the compound is tested by Ferric reducing antioxidant power (FRAP) test and safety of the compound is identified through acute toxicity by [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Moreover, activities of superoxide dismutase (SOD), catalase (CAT), levels of prostaglandins E2 (PGE2) and also malondialdehyde (MDA) are determined. Results Antioxidant activity of CNBP was approved via FRAP assay. Vast shallow hemorrhagic injury of gastric glandular mucosa was observed in the ulcer group compared to the CNBP-treated animals. Histological evaluations confirmed stomach epithelial defense effect of CNBP with drastic decrease of gastric ulceration, edema and leucocytes penetration of submucosal stratum. Immunostaining exhibited over-expression in HSP70 protein in CNBP-treated groups compared to that of the ulcer group. Also, gastric protein analysis showed low levels of MDA, PGE2 and high activity of SOD and CAT. Conclusions CNBP with noticeable antioxidant property showed gastroprotective activity in the testing rodents via alteration of HSP70 protein expression. Also, antioxidant enzyme activities which were changed after treatment with CNBP in the animals could be elucidated as its gastroprotective properties

Cinnamomum cassia Suppresses Caspase-9 through Stimulation of AKT1 in MCF-7 Cells but Not in MDA-MB-231 Cells.

BACKGROUND:Cinnamomum cassia bark is a popular culinary spice used for flavoring and in traditional medicine. C. cassia extract (CE) induces apoptosis in many cell lines. In the present study, particular differences in the mechanism of the anti-proliferative property of C. cassia on two breast cancer cell lines, MCF-7 and MDA-MB-231, were elucidated. METHODOLOGY/PRINCIPAL FINDINGS:The hexane extract of C. cassia demonstrated high anti-proliferative activity against MCF-7 and MDA-MB-231 cells (IC50, 34 ± 3.52 and 32.42 ± 0.37 μg/ml, respectively). Oxidative stress due to disruption of antioxidant enzyme (SOD, GPx and CAT) activity is suggested as the probable cause for apoptosis initiation. Though the main apoptosis pathway in both cell lines was found to be through caspase-8 activation, caspase-9 was also activated in MDA-MB-231 cells but suppressed in MCF-7 cells. Gene expression studies revealed that AKT1, the caspase-9 suppressor, was up-regulated in MCF-7 cells while down-regulated in MDA-MB-231 cells. Although, AKT1 protein expression in both cell lines was down-regulated, a steady increase in MCF-7 cells was observed after a sharp decrease of suppression of AKT1. Trans-cinnamaldehyde and coumarin were isolated and identified and found to be mainly responsible for the observed anti-proliferative activity of CE (Cinnamomum cassia). CONCLUSION:Activation of caspase-8 is reported for the first time to be involved as the main apoptosis pathway in breast cancer cell lines upon treatment with C. cassia. The double effects of C. cassia on AKT1 gene expression in MCF-7 cells is reported for the first time in this study

CE induces apoptosis in breast cancer cells.

<p>(A) MCF-7, MDA-MB-231 cells were stained with Annexin V/PI and subjected to flow cytometric analysis. The four quadrants represent living cells (Annexin V-PI-), early apoptotic (Annexin V+PI-), late apoptotic (Annexin+PI+) or necrotic (Annexin V-PI+) stages. (B) Percentage level of apoptotic cells when the cells were treated with different concentrations of CE. Values shown are percentages of each quadrant. *P<0.05, in comparison to control. The development of color was examined by bright field microscopy. (Magnification 200X). Z-VAD-FMK (caspase-8 inhibitor) was used as a negative control.</p

The effect of CE on gene expression in MCF-7 and MDA-MB-231 cells.

<p>Both cell lines were treated with the IC₅₀ concentration (35 μg/ml) of CE for 24 h. Total RNA was extracted and cDNA was synthesized using commercial kits. Real-time RT-PCR was performed and the relative expression of the genes was calculated using the delta-delta Ct method and normalized with 18S eukaryotic rRNA. Results are expressed as fold variation over carrier control (blank). Results are expressed as mean ± standard deviation. Statistical significance was calculated based on the mean ΔCt values by the Student’s <i>t</i> test. *Indicates significant differences from untreated cells (p < 0.05).</p

The Caspase-Glo luminoscence of MCF-7 and MDA-MB-231 cells treated with CE, without treatment, and treated with caspase inhibitor.

<p>Cells were treated with the IC₅₀ concentration (35 μg/ml) of CE at varying time points (2, 8, 16, 24, 28 h). (A) The Caspase-Glo luminoscence of MCF-7 and MDA-MB-231cells treated with CE, without treatment and treated with caspase inhibitor. (B) The fold change in caspase activity of MCF-7 and (C) MDA-MB-231cells treated with CE. Results are expressed as mean ± standard deviation. P < 0.05 compared to the control (without extract) as tested by the Student’s <i>t</i>-test.</p

Antiproliferative activity of CE, <i>trans</i>-cinnamaldehyde and coumarin in MDA-MB-231 and MCF-7 cells.

<p>(A) The % inhibition when MDA-MB-231 and MCF-7 cells were exposed to various concentrations of the CE. (B) The % inhibition when MDA-MB-231 cells were exposed to various concentrations of <i>trans</i>-cinnamaldehyde and coumarin. (C) The % inhibition when MCF-7 cells were exposed to the various concentrations of <i>trans</i>-cinnamaldehyde and coumarin. Results are expressed as mean ± std. dev. (n = 3). IC₅₀ is defined as the concentration of extract that inhibited 50% of cell proliferation. ND = not detected.</p