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Research in Pharmaceutical Sciences - Volume:11 Issue: 5, Oct 2016

Research in Pharmaceutical Sciences
Volume:11 Issue: 5, Oct 2016

  • تاریخ انتشار: 1395/08/05
  • تعداد عناوین: 10
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  • Samaa Alrushaid, Neal M. Davies, Stephanie E. Martinez, Casey L. Sayre Pages 355-365
    Liquiritigenin is a chiral flavonoid present in plant based food, nutraceuticals, and traditional medicines. It is also an important ingredient present in licorice. The purpose of this study is to explore the pharmacological activity of racemic liquiritigenin utilizing several in vitro assays with relevant roles in colon cancer and diabetes. Where possible, the pure enantiomers were tested to identify the stereospecific contribution to the activity. In vitro antioxidant, anticancer, anti-inflammatory activities (cyclooxygenase inhibition), antidiabetic activities (alpha-amylase and alpha-glucosidase inhibition) as well as cytochrome P450 (CYP450) inhibitory activities were assessed. Racemic liquiritigenin demonstrated a dose-dependent inhibition of alpha-amylase enzyme whereas its pure enantiomers did not. Racemic liquiritigenin showed moderate antiproliferative activity on a HT-29 (human colorectal adenocarcinoma) cancer cell line that was dose-dependent and potent inhibitory effects on the cyclooxygenase-2 enzyme. The flavonoid did not inhibit the activity of cytochrome CYP2D6 over the concentration range studied but was a potent antioxidant. The current study demonstrated the importance of understanding the stereospecific pharmacological effects of liquiritigenin enantiomers in alpha-amylase inhibition.
    Keywords: Liquiritigenin, Flavonoid, Chiral, Stereospecific, Pharmacology
  • Mina Mirian, Razieh Taghizadeh, Hossein Khanahmad, Mansour Salehi, Ali Jahanian, Najafabadi, Hojjat Sadeghi, Aliabadi, Shirin Kouhpayeh Pages 366-373
    Hepatitis B virus (HBV) is considered as a global health concern and hepatitis B surface antigen (HBsAg) is the most immunogenic protein of HBV. The purpose of this study was to evaluate the expression of HBsAg on the cell surface of human embryonic kidney cell line (HEK293T). After transformation of expression vector pcDNA/HBsAg to E.coli TOP10F’, plasmid was extracted and digested with BglII. Afterwards, the linearized vector was transfected to cells and treated with hygromycin B for 5 weeks to expand the resulted clonies. The permanent expression of HBsAg followed by flow cytometry uptill now about one year. Genomic DNA was extracted from transfected cells and the existence of HBsAg gene was assessed by PCR. Real-time RT-PCR was utilized to measure the expression at the RNA level and flow cytometery was carried out to assess protein expression. Insertion of HBsAg cDNA in HEK293T genome was confirmed by PCR. The results of real-time RT-PCR illustrated that each cell expresses 2275 copies of mRNA molecule. Flow cytometry showed that compared with negative control cells, 99.9% of transfected cells express HBsAg on their surface. In conclusion, stable expression of hepatitis B surface antigen on the membrane of HEK293T provides an accurate post-translational modification, proper structure, and native folding in contrast with purified protein from prokaryotic expression systems. Therefore, these exposing HBsAg cells are practical in therapeutic, pharmaceutical, and biological sets of research.
    Keywords: Hepatitis B, HBsAg, Recombinant HEK293T cell
  • Naglaa Mohamed El, Lakkany, Sayed Hassan Seif El, Din, Abdel, Nasser Abdel, Aal Sabra, Olfat Ali Hammam, Fatma Abdel, Latif Ebeid Pages 374-382
    Non-alcoholic fatty liver disease (NAFLD) is a burgeoning health problem that affects 1/3 of the adult population and an increasing number of children in developed countries. Oxidative stress and insulin resistance are the mechanisms that seem to be mostly involved in its pathogenesis. This study was conceived in a NAFLD rat model to evaluate the efficacy of both metformin (MTF) and N-acetylcysteine (NAC) with dietary control on biochemical and histologic liver manifestations. Rats were classified into nine groups; normal (I), NAFLD-induced by feeding high-fat diet (HFD; II) for 12 weeks, NAFLD switched to regular diet (RD; III), NAFLD-HFD or -RD treated with MTF in a dose of 150 mg/kg (IV, V), NAC in a dose of 500 mg/kg (VI, VII) or MTF㐀 (VIII, IX) respectively for 8 weeks. After 20 weeks, the rats in group II showed notable steatosis, lobular inflammation, fibrosis accompanied with elevated (P
    Keywords: NAFLD, Metformin, N, acetylcysteine, Oxidative stress, Adipocytokines, TNF
  • Leila Safaeian, Seyyed Ebrahim Sajjadi, Shaghayegh Haghjooy Javanmard, Hossein Montazeri, Fariba Samani Pages 383-389
    Melissa officinalis L. is a medicinal plant with a large variety of pharmacological effects and traditional applications. This study aimed to evaluate the protective and antioxidant activities of the extract of M. officinalis aerial parts on human umbilical vein endothelial cells (HUVECs) under oxidative stress induced by H2O2. Cells were incubated with H2O2 (0.5 mM, 2 h) after pretreatment with M. officinalis extract (25-500 µg/mL). Cell viability was evaluated by 3-(4, 5- Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The concentration of hydroperoxides and ferric reducing antioxidant power (FRAP) were measured in intra- and extra-cellular fluids. Pretreatment of HUVECs with M. officinalis extract at the concentrations of 100-500 µg/mL improved the cell viability after exposure to H2O2 significantly. It also decreased hydroperoxides concentration and increased FRAP value in both intra- and extra-cellular fluids. The results revealed antioxidant and cytoprotective effects of M. officinalis against H2O2-induced oxidative stress in HUVECs. Due to the valuable antioxidant activity, this plant extract may have potential benefits for the prevention of cardiovascular diseases associated with oxidative stress.
    Keywords: Melissa officinalis L., HUVECs, Oxidative stress, Antioxidant
  • Saeedeh Alsadat Moosavirad, Mohammad Rabbani, Mohammad Sharifzadeh, Ali Hosseini, Sharifabad Pages 390-396
    Lead belongs to the heavy metal group and is considered as an environmental contaminant. Acute or chronic contact to lead can change the physiological function of human organs. One of the most important disorders following the lead exposure is neurotoxicity. Lead neurotoxicity consists of the neurobehavioral disturbances like cognitive impairment. The aim of the current study is to evaluate the possible protective effect of vitamin C (Vit C), vitamin B12 (Vit B12), omega 3 (ω-3), or their combination on the lead-induced memory disorder. Adult wistar rats were orally administered Vit C (120 mg/kg/day) or Vit B12 (1 mg/kg/day) or ω-3 (1000 mg/kg/day) or their combination for 3 weeks in groups of 7 animals each. Then lead acetate (15 mg/kg/day) was injected intraperitoneally for one week to all pretreated animals. The control group received normal saline as a vehicle while the positive control for cognitive impairment received just lead acetate. At the end of treatments animal memories were evaluated in Object Recognition Task. The results showed, although 15 mg/kg lead acetate significantly declines the memory-evaluating parameters, pretreatment with Vit C, Vit B12, ω-3, or their combination considerably inverted the lead induced reduction in discrimination (d2) index (P
    Keywords: Vitamin C, Vitamin B12, Omega, 3, Lead, Memory impairment, Object Recognition Task
  • Inemesit Okon Ben, Eric Woode, George Asumeng Koffuor, Emmanuel Akomanin Asiamah Pages 397-404
    Effects of petroleum ether and ethanolic extracts of Trichilia monadelpha stem bark (PEE and EAE) on compound 48/80-induced systemic and passive anaphylaxis were determined. Survival rate, extravasation, degranulation of mast cells, and secretion of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured after pre-treatment with extracts (10-100 mg/kg) and disodium chromoglycate (2.5-250 µg/kg) and induction of anaphylaxis in C57BL/6 mice or Sprague-Dawley rats with compound 48/80. Histopathological assessments were made from skin biopsies of rats. Data was analyzed by Kaplan-Meier Survival Log-Rank Analysis, or One-way ANOVA and Holm-Sidak’s post hoc test. PEE and EAE inhibited (P ≤ 0.0001) tremors in systemic anaphylaxis passive cutaneous anaphylactic reactions and extravasation, stabilized or prevented (P ≤ 0.001-0.0001) mast cell degranulation, and inhibited (P ≤ 0.001-0.0001) TNF-α and IL-6 secretion. Per the findings, PEE and EAE of T. monadelpha have exhibited substantial anti-anaphylactic and anti-inflammatory property (with PEE performing better) which substantiates its use traditionally in management of allergies and other inflammatory disorders.
    P
    Keywords: Mast cells degranulation, Trichilia monadelpha, Interleukin, 6, TNF, α Anaphylaxis
  • Ahmad Movahedian, Behzad Zolfaghari, Mehrzad Mirshekari Pages 405-411
    Antioxidant activity of Peucedanum pastinacifolium Boiss. & Hausskn aerial part hydroalcoholic extract (HAE) and polyphenolic extract (PPE) as well as their total phenolic and flavonoid contents were studied. Phenolic and flavonoid contents were respectively estimated as gallic acid and quercetin equivalents. The in vitro antioxidant activity of two extracts of P. pastinacifolium were evaluated by radical scavenging of 1,1-diphenyl-2-picryl hydrazyl radical (DPPH), chelating activity on ferrous ions, or ferric reducing antioxidant power (FRAP) assay. In addition, the in vivo antioxidant activity of hydroalcoholic extract was measured by FRAP assay. Total phenolic contents of PPE and HAE were 117.1 ± 6.2 and 44.3 ± 1.7 mg/g, respectively. Total flavonoid content of PPE (43.4 ± 2.1 mg/g) was found to be higher than that of HAE (8.0 ± 1.5 mg/g). In DPPH radical scavenging assay, HAE and PPE showed fifty percent inhibitory concentration (IC50) values of 469.4 ± 9.3 µg/mL and 128.2 ± 5.5 µg/mL, respectively. Iron chelating activity assays indicated IC50 values of 657.5 ± 13.2 µg/mL and 735.4 ± 16.1 µg/mL for HAE and PPE as opposed to ethylenediamine tetra-acetic acid (EDTA) being 16.5 ± 0.8 µg/mL. PPE exhibited greater FRAP value (154.0 ± 1.8 µM) as compared with that of HAE being 69.3 ± 1.4 µM. In animal study, HAE showed a significant (p
    Keywords: Peucedanum pastinacifolium, Antioxidant, Reducing power, Free radicals, Chelating activity
  • Soheil Yousefi, Mojtaba Tahmoorespur, Mohammad Hadi Sekhavati Pages 412-418
    Brucellosis is a well-known domestic animal infectious disease, which is caused by Brucella bacterium. The outer membrane protein 25 kDa (Omp25) gene plays an important role in simulating of TNF-α, IFN-α, macrophage, and cytokines cells. In the current study molecular cloning and expression analysis of Omp25 gene for designing a subunit vaccine against Brucella was investigated. Amplifying the full length of candidate gene was performed using specific primers. Sub-cloning of this gene conducted using pTZ57R/T vector in TOP10F`strain of Escherichia coli(E.coli) as the host. Also, pET32(a) vector used for expression in BL21 (DE3) strain of E.coli. Omp25 gene with 642 bp size was amplified and cloned successfully. The expression results were confirmed by sequencing and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analyses which showed 42 kDa protein band correctly. Also, phylogenic analysis showed this gene has a near genetic relation with other Brucella strains. According to our results we can propose this gene as a candidate useful for stimulation of cell-mediated and humoral immunity system in future study.
    Keywords: Brucella melitensis Rev1, Omp25, Phylogenic analysis, Gene expression
  • Geeta B. Kharadi, Kaksha J. Patel, Bhargav M. Purohit, Seema N. Baxi, C.B. Tripathi Pages 419-427
    To investigate the cardioprotective potential of the aqueous extract of Allium cepa Linn. bulb in isoprenaline-induced myocardial injury in Wistar albino rats. In vitro total phenolic, total flavonoid content and 2, 2’-diphenyl-1-picrylhydrazyl hydrate radical scavenging activity was measured. Isoprenaline-induced myocardial injury model was used to evaluate in vivo effect of aqueous extract of A. cepa in Wistar albino rats. Seventy two rats were randomly divided in 6 groups. Rats were treated with A. cepa 400 mg/kg and 800 mg/kg doses for 30 days and myocardial injury was produced by subcutaneous injection of isoprenaline (ISO) 85 mg/kg on day 28 and 29. Carvedilol 1 mg/kg for 30 days served as active control. Electrocardiogram parameters, cardiac injury markers, oxidative stress markers and histopathological changes were evaluated in each group and compared using appropriate statistical tests. In vitro evaluation of aqueous extract of A. cepa showed significant antioxidant property. ISO produced significant myocardial injury as compared to normal control group (P 0.05). The aqueous extract of A. cepa 400 mg/kg was found to be cardioprotective against myocardial injury while A. cepa 800 mg/kg did not show significant cardioprotective activity. So, we presume that A. cepa might be effective within certain dose range only.
    Keywords: Allium cepa, Cardioprotective, Isoprenaline, Carvedilol, Myocardial infarction
  • Fatemeh Shafiee, Mohammad Rabbani, Mahdi Behdani, Ali Jahanian, Najafabadi Pages 428-434
    The aim of this study was to produce a recombinant protein consisting of the catalytic and translocation domains of diphtheria toxin for its later application as a vaccine candidate against Corynebacterium diphtheria. To achieve this goal, at first, the amino acid sequence of DT386 was used for prediction of T and B cell epitopes using on-line servers. The DT386 coding sequence was synthesized and subcloned into the NcoI and XhoI sites of pET28a plasmid and recombinant pET28a plasmid was used to transform Escherichia coli BL21 (DE3) host cells. Afterwards, recombinant cells were selected and subjected to induction of expression by 1 mM isopropyl β-D-1-thiogalactopyranoside, (IPTG). Expression of the desired protein was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting, and finally, the recombinant protein was purified using nickel affinity chromatography. The results of epitope prediction using on-line servers established the ability of DT386 for stimulation of immune system against diphtheria toxin. Restriction digestion of the recombinant plasmids using NcoI and XhoI enzymes confirmed the fidelity of cloning by producing a band of about 1200 bp. SDS-PAGE analysis following induction of expression and also purification step confirmed the expression of the desired protein by showing a band of about 45 kDa. In addition, Western blot analysis using anti-6X-His antibody confirmed the identity of the expected protein. In conclusion, in the present study we amplified and cloned the coding sequence of DT386 fragment, followed by its expression by E. coli BL21 (DE3) cells. Then, the expressed protein was purified and will be used for later studies of evaluation of its immunogenic properties.
    Keywords: Diphtheria toxin, DT386, Vaccine candidate, Immunoinformatics