scholarly journals Comparative Proteomic Analysis of Differential Proteins in Response to Aqueous Extract of Quercus infectoria Gall in Methicillin-Resistant Staphylococcus aureus

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Radhiah Khairon ◽  
Noraziah Mohamad Zin ◽  
Mariati Abdul Rahman ◽  
Dayang Fredalina Basri

The aim of this study is to analyze the differential proteins in MRSA ATCC 33591 treated with aqueous extract from Q. infectoria gall. Protein extracts were obtained from MRSA cells by sonication and were separated by 2D polyacrylamide gels. Protein spots of interest were extracted from the gels and identified using LC-ESI-QTOF MS. The concentration of Q. infectoria extract used for 2D-gel electrophoresis was subinhibitory concentration. Minimum inhibitory concentration (MIC) value of the extract against MRSA was 19.50 μg/mL with bacteriostatic action at 1x MIC from time-kill assay. However, the extract exhibited dose-dependent manner and was bactericidal at 4x MIC with more than 3 log10 CFU/mL reduction at 4 h. 2D-GE map showed that 18 protein spots were upregulated and another six were downregulated more than twofold (p<0.05) after treatment with subinhibitory concentration. Out of six proteins being downregulated, four proteins were identified as ferritin and catalase, branched-chain alpha-keto acid dehydrogenase subunit E2, and succinyl-CoA ligase [ADP-forming] subunit beta. Seven upregulated proteins which have been successfully identified were 3-hydroxyacyl-CoA dehydrogenase, NAD binding domain protein, formate C-acetyltransferase, 3-hydroxyacyl-[acyl-carrier-protein] dehydratase FabZ, NAD dependent epimerase/dehydratase family protein, and phosphopantothenoyl cysteine decarboxylase. It is postulated that the main mechanism of aqueous extract from gall of Q. infectoria was most likely involved in energy metabolism and protein stress.

Author(s):  
Retno Widyowati ◽  
Suciati Suciati ◽  
Dewi Melani Haryadi ◽  
Hsin-I Chang ◽  
IPG Ngurah Suryawan ◽  
...  

Abstract Objectives Glucocorticoid-induced osteoporosis (dexamethasone) is a primary cause of secondary osteoporosis by the decreasing formation and increasing resorption activities. Previously, the in vitro study showed that 70% ethanol and aqueous extract of deer antler have increased alkaline phosphatase in osteoblast cell that known as marker of bone formation. The mind of this study is to analyze the effect of deer antlers in increasing the bone trabecular density of osteoporosis-induced male mice. Methods This study used a post-test control group design. A total of 54 healthy male mice were randomly divided to nine groups, i.e., healthy control, osteoporotic, positive control, 70% ethanol (4, 8, and 12 mg/kg BW), and aqueous extracts (4, 8, and 12 mg/kg BW) of deer antler groups. All of the interventions were given 1 mL of test sample for 4 weeks orally. The bone densities were determined using histomorphometry by Image J and Adobe Photoshop. The statistical data were performed using SPSS 23 and statistical significance was set at p<0.05. Results The results showed that alendronate group, 70% ethanol, and aqueous extract groups increased bone density and calcium levels in serum (p<0.05) compared to osteoporotic group in dose dependent manner. It indicated that 70% ethanol and aqueous extract of deer antler stimulating bone turnover and aqueous extract showed the highest. Conclusions Dexamethasone induction for 4 weeks caused osteoporotic mice and the administration of 70% ethanol and aqueous extracts of deer antler from East Kalimantan increased trabecular bone density and calcium levels in dose dependent manner.


2021 ◽  
Vol 17 ◽  
Author(s):  
Brahim Asseli ◽  
Reguia Mahfoudi ◽  
Amar Djeridane ◽  
Mohamed Yousfi

Background: Research on medicinal plant antioxidants has emerged as a potential therapeutic to prevent free radical generated damage in the human body. Hammada elegans Botsch (popularly known as “Ajram”) is a xerophytic plant widely found in Laghouat region, but there are only a few reports about the biological or chemical properties of these species. Hence, the aim of this study is to investigate the antioxidant and the antihemolytic activities of hexanic, acetonic, methanolic and aqueous extracts of aerial parts of Algerian Hammada elegans Botsch by employing different in vitro assay systems. Methods: The total phenolic content, the flavonoid content and the condensed tannin amount were analyzed using Folin-Ciocalteu, aluminum chloride and vanillin assays, respectively. The in vitro antioxidant capacity of extracts was assessed by CUPRAC, iron chelating, ABTS•+and antihemolytic assays, and was expressed as EC50 values. Results: Among the analyzed extracts, the aqueous extract had the highest phenolic, flavonoid and tannin contents. Also, this extract displayed the highest antioxidant capacities compared to the other extracts and standards. Its EC50 value for ABTS radical-scavenging activity was 0.265 ± 0.003 mg/L. Moreover, this extract showed high iron (II) chelating ability (EC50 = 0.958 ± 0.001 mg/L), and good antioxidant activity in the cupric ion reducing activity (CUPRAC) in a concentration dependent manner (EC50 were 0.709 ± 0.002 mg/L). Additionally, this extract had the best antihemolytic activity against AAPH-induced hemolysis (EC50=0.090 ± 0.004 mg/L). Conclusion: Our study revealed that the aqueous extract of Hammada elegans Botsch, is a potential source of antioxidants which possess a high protective effect of membrane against free radical.


Author(s):  
K. Amala ◽  
R. Ilavarasan ◽  
R. Arunadevi ◽  
S. Amerjothy

<p><strong>Objective: </strong>The plant of <strong><em>Epaltes</em></strong><strong> <em>divaricata </em>(L.) </strong>Cass.<strong> Traditionally used for jaundice. </strong>The present work aimed to investigate the hepatoprotective activity of alcohol and aqueous extract of the whole plant against paracetamol-induced hepatotoxicity in rats to substantiate its traditional use.</p><p><strong>Methods: </strong>The alcohol and aqueous (200 and 400 mg/kg) extract of <em>Epaltes divaricata</em> prepared by cold maceration were administered orally to the animals with hepatotoxicity induced by paracetamol (1000 mg/kg). Silymarine (40 mg/k) was given as reference standard. Hepatoprotective activity was assessed by estimating marker enzymes and by histopathological studies.</p><p><strong>Results: </strong>Both alcohol and aqueous (200 and 400 mg/kg) extract treatment significantly restored the paracetamol-induced elevations in levels of serum enzymes aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphate (ALP) and total bilirubin in a dose-dependent manner. Histopathological examination revealed that the treatment attenuated the paracetamol-induced damage to the liver. The hepatoprotective effect of both extracts was comparable to that of the standard hepatoprotective agent, silymarin.</p><p><strong>Conclusion: </strong>The alcohol and aqueous extract of <em>E. divaricata</em> exhibited hepatoprotective effect against paracetamol-induced liver damage in rats. This study also validated their traditional medicinal use in jaundice.</p>


Author(s):  
Okoye Ngozi Franca ◽  
Ikiriko, Favour Ibiwari

Aim: This study was aimed at investigating the effects of aqueous extracts of both Nigella sativa and Piper guineense on the liver enzymes; alanine amino transferase (ALT), aspartate amino transferase (AST) and alkaline phosphatase (ALP). Also the effect of Nigella sativa and Piper guineense extracts on the histology of the liver of Wistar rat was also studied. Materials and Methods: A total of twenty five Wistar rats were used for the study. The animals were grouped into five groups, each having five animals. They were induced with sucrose and margarine to cause high sugar levels and hyperlipidemia respectively except the positive control group which was fed normal feed. The groups were: the positive control group, the negative control group which were induced without treatment, the uziza leaf group which were induced and were treated with 2 ml of aqueous extract of uziza leaf, the black seed group which were induced and were treated with 2 ml of aqueous extract of black seed, and the black seed and uziza group which were induced and were treated with 2ml of aqueous extract of black seed and 2 ml of aqueous extract of uziza leaf.  Results: The result showed that the extracts decreased the ALT and AST and ALP activities in the rats in a time dependent manner with highest decrease obtained on the third week of treatment with the extracts.  The ALT activity (U/L) on the third week of treatment showed for the, negative control (64.48 ± 0.22), uziza leaf (28.82 ± 0.12), black seed (32.65 ± 0.02), black seed and uziza leaf (16.04 ± 0.02) (p≤0.05). The decrease in activity for AST levels (U/L) on the third week of treatment, showed for the negative control (58.00 ± 0.02), uziza leaf (11.00 ± 0.01), black seed (12.00 ± 0.02), black seed and uziza leaf (8.00 ± 0.02). Conclusion: It can be concluded that both uziza leaf and black seed have hepatoprotective effect on the liver.


2017 ◽  
Vol 8 (4) ◽  
pp. 525
Author(s):  
Chandra Kishore Tyagi ◽  
Deenanath Jhade ◽  
Sunil Kumar Shah

<p>The study evaluated anticoagulant properties of the aqueous extract of <em>Cestrum nocturnum</em> using aPTT-Activated Partial Thromboplastin Time, PT- Prothrombin Time &amp; TT-Thrombin Time as standard procedures.</p><p>For <em>in vitro</em> coagulation assays, aqueous extract of plant prolonged APTT, TT, and PT clotting times in a dose-dependent manner (Table 7). It prolonged APTT clotting time from 45 ± 2 (2mg/mL) to 82.2 ± 2.63s (10mg/mL), PT clotting time from 20.4 ± 1.49 (2mg/mL) to 31.4 ± 2.15s (10mg/mL), and TT clotting time from 9.2 ± 1.16 (2mg/mL) to 17.4 ± 1.01s (10mg/mL) at the concentration of 2 to 10mg/mL. Heparin prolonged APTT and PT clotting times more than 111.8s and 40.8s, respectively, at a concentration of 1 IU/mL. Heparin prolonged TT clotting times more than 20.6s at a concentration of 1 IU/mL.</p><p>The phytochemical screening of the plant confirm the presence of saponin in the water and ethanolic extract, Alkaloid in all the extract except hexane extract, tannin in water, ethanol and methanol extract, amino acid in water and ethanolic extract, carbohydrate in water and methanolic extract and triterpenoids and glycoside were absent in all the extracts. The results demonstrated that the aqueous extract of <em>Cestrum nocturnum</em> possesses pharmacologically active anticoagulant principles that could be isolated and evaluated for clinical or physiological purposes.</p>


2018 ◽  
Vol 6 (7) ◽  
pp. 1206-1214
Author(s):  
Ghassan Hamdan Jameel ◽  
Ali Ibrahim Ali AL-Ezzy ◽  
Ibrahim H. Mohammed

OBJECTIVES: To evaluate the Immunomodulatory, apoptosis induction and antitumor effects of aqueous and methanolic extracts of Calvatia craniiformis regarding the size of tumour mass, caspase-8 expression and apoptotic index (AI%) in mice transfected with murine hepatocellular carcinoma cell line (H22) as an experimental therapeutic system for human hepatocellular carcinoma.MATERIAL AND METHODS: Forty-eight Balb/C albino mice were transfected in legs with H22 cells. Tumour size was measured twice a week. Caspase-8 protein expression and apoptotic index determination evaluated by Immunohistochemistry.RESULTS: Tumor size significantly differed between the two groups of mice transfected with H22 cells; the first was treated with C. craniiformis aqueous extract (0.3, 0.6, 1.2) mg/kg and the second group was treated with C. craniiformis methanolic extract (0.25, 0.5, 1.0) mg/kg compared with control group. The inhibitory activity of aqueous and methanolic extracts was dose and duration dependent. The size of the tumour mass was reduced up to 87.9% when treated with 1.2 mg/kg aqueous extract and 1 mg/kg for methanolic extract. Caspase-8 expression was increased in a dose-dependent manner among H22 bearing mice treated with C. craniiformis aqueous extract (0.3, 0.6, 1.2) mg/kg. At 0.3 mg/kg, the intensity of expression was strong in (33.33%) and very strong in (66.67%). While at 0.6 mg/kg and 1.2 mg/kg the intensity of expression was strong in (33.33%) and very strong in (100%) with a significant difference (P ≤ 0.001). H22 bearing mice treated with (0.25, 0.5, 1.0) mg/kg C. craniiformis methanolic extract shows increased caspase-8 expression in a dose-dependent manner. At 0.25 mg/kg, the intensity of expression was strong in (33.33%) and very strong in (66.67%). While at 0.5 mg/kg, the intensity of expression was strong in (33.33%) and very strong in (100%). At 1.0 mg/kg, the intensity of expression was strong in (16.67%) and very strong in (83.33%) with significant difference (P ≤ 0.001). AI% of H22 bearing mice treated with C. craniiformis aqueous and methanolic extracts were significantly increased (P ≤ 0.05) compared with the untreated control group. No significant difference was reported in AI% between aqueous and methanolic extracts treated groups.CONCLUSIONS: Extracts of C. craniiformis were highly efficient in tumour growth inhibition, causing a reduction in the tumour size clinically and increase the expression of caspase-8 gene product in tumour tissue, causing increase apoptotic index of H22 cells taken from the legs of inoculated mice leading to loss of legs due to bone necrosis. Antitumor activity of C. craniiformis aqueous, and the methanolic extract was dose and duration dependent.


2020 ◽  
Vol 13 (10) ◽  
pp. dmm047134
Author(s):  
Leonie Drews ◽  
Marcel Zimmermann ◽  
Philipp Westhoff ◽  
Dominik Brilhaus ◽  
Rebecca E. Poss ◽  
...  

ABSTRACTAstrocyte dysfunction is a primary factor in hepatic encephalopathy (HE) impairing neuronal activity under hyperammonemia. In particular, the early events causing ammonia-induced toxicity to astrocytes are not well understood. Using established cellular HE models, we show that mitochondria rapidly undergo fragmentation in a reversible manner upon hyperammonemia. Further, in our analyses, within a timescale of minutes, mitochondrial respiration and glycolysis were hampered, which occurred in a pH-independent manner. Using metabolomics, an accumulation of glucose and numerous amino acids, including branched chain amino acids, was observed. Metabolomic tracking of 15N-labeled ammonia showed rapid incorporation of 15N into glutamate and glutamate-derived amino acids. Downregulating human GLUD2 [encoding mitochondrial glutamate dehydrogenase 2 (GDH2)], inhibiting GDH2 activity by SIRT4 overexpression, and supplementing cells with glutamate or glutamine alleviated ammonia-induced inhibition of mitochondrial respiration. Metabolomic tracking of 13C-glutamine showed that hyperammonemia can inhibit anaplerosis of tricarboxylic acid (TCA) cycle intermediates. Contrary to its classical anaplerotic role, we show that, under hyperammonemia, GDH2 catalyzes the removal of ammonia by reductive amination of α-ketoglutarate, which efficiently and rapidly inhibits the TCA cycle. Overall, we propose a critical GDH2-dependent mechanism in HE models that helps to remove ammonia, but also impairs energy metabolism in mitochondria rapidly.


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Zhihong Lin ◽  
Danni Zhu ◽  
Yongqing Yan ◽  
Boyang Yu ◽  
Qiujuan Wang ◽  
...  

Oxidative stress is involved in the pathogenesis of ischemic neuronal injury. A Chinese herbal formula composed ofPoria cocos(Chinese name:Fu Ling),Atractylodes macrocephala(Chinese name:Bai Zhu) andAngelica sinensis(Chinese names:Danggui, Dong quai, Donggui; Korean name:Danggwi) (FBD), has been proved to be beneficial in the treatment of cerebral ischemia/reperfusion (I/R).This study was carried out to evaluate the protective effect of FBD against neuronal oxidative stressin vivoandin vitro. Rat I/R were established by middle cerebral artery occlusion (MCAO) for 1 h, followed by 24 h reperfusion. MCAO led to significant depletion in superoxide dismutase and glutathione and rise in lipid peroxidation (LPO) and nitric oxide in brain. The neurological deficit and brain infarction were also significantly elevated by MCAO as compared with sham-operated group. All the brain oxidative stress and damage were significantly attenuated by 7 days pretreatment with the aqueous extract of FBD (250 mg kg−1, p.o.). Moreover, cerebrospinal fluid sampled from FBD-pretreated rats protected PC12 cells against oxidative insult induced by 0.2 mM hydrogen peroxide, in a concentration and time-dependent manner (IC5010.6%, ET501.2 h). However, aqueous extract of FBD just slightly scavenged superoxide anion radical generated in xanthine–xanthine oxidase system (IC502.4 mg ml−1) and hydroxyl radical generated in Fenton reaction system (IC503.6 mg ml−1). In conclusion, FBD was a distinct antioxidant phytotherapy to rescue neuronal oxidative stress, through blocking LPO, restoring endogenous antioxidant system, but not scavenging free radicals.


Planta Medica ◽  
2019 ◽  
Vol 85 (11/12) ◽  
pp. 997-1007 ◽  
Author(s):  
Katerina Gioti ◽  
Anastasia Papachristodoulou ◽  
Dimitra Benaki ◽  
Sophia Havaki ◽  
Apostolos Beloukas ◽  
...  

AbstractSilymarin-enriched extract (SEE) is obtained from Silybum marianum (Asteraceae). Doxorubicin (DXR) is a widely used chemotherapeutical yet with severe side effects. The goal of the present study was to assess the pharmacologic effect of SEE and its bioactive components silibinin and silychristine when administrated alone or in combination with DXR in the human prostate cancer cells (PC-3). PC-3 cells were treated with SEE, silibinin (silybins A and B), silychristine, alone, and in combination with DXR, and cell proliferation was assessed by the MTT assay. Cell cycle, apoptosis, and autophagy rate were assessed by flow cytometry. Expression levels of autophagy-related genes were quantified by qRT-PCR, ELISA and western blot while transmission electron microscopy was performed to reveal autophagic structures. Finally, NMR spectrometry was used to identify specific metabolites related to autophagy. SEE inhibited PC-3 cell proliferation in a dose-dependent manner while the co-treatment (DXR-SEE) revealed an additive cytotoxic effect. Cell cycle, apoptosis, and autophagy variations were observed in addition to altered expression levels of autophagy related genes (LC3, p62, NBR1, Beclin1, ULK1, AMBRA1), while several modifications in autophagic structures were identified after DXR-SEE co-treatment. Furthermore, treated cells showed a different metabolic profile, with significant alterations in autophagy-related metabolites such as branched-chain amino acids. In conclusion, the DXR-SEE co-treatment provokes perturbations in the autophagic mechanism of prostate cancer cells (PC-3) compared to DXR treatment alone, causing an excessive cell death. These findings propose the putative use of SEE as an adjuvant cytotoxic agent.


2020 ◽  
Vol 21 (22) ◽  
pp. 8493
Author(s):  
Hiroki Kazama ◽  
Shu-ichiro Kashiwaba ◽  
Sayaka Ishii ◽  
Keiko Yoshida ◽  
Yuta Yatsuo ◽  
...  

Cell division is essential for the maintenance of life and involves chromosome segregation and subsequent cytokinesis. The processes are tightly regulated at both the spatial and temporal level by various genes, and failures in this regulation are associated with oncogenesis. Here, we investigated the gene responsible for defects in cell division by using murine temperature-sensitive (ts) mutant strains, tsFT101 and tsFT50 cells. The ts mutants normally grow in a low temperature environment (32 °C) but fail to divide in a high temperature environment (39 °C). Exome sequencing and over-expression analyses identified Diaph3, a member of the formin family, as the cause of the temperature sensitivity observed in tsFT101 and tsFT50 cells. Interestingly, Diaph3 knockout cells showed abnormality in cytokinesis at 39 °C, and the phenotype was rescued by re-expression of Diaph3 WT, but not Diaph1 and Diaph2, other members of the formin family. Furthermore, Diaph3 knockout cells cultured at 39 °C showed a significant increase in the level of acetylated α-tubulin, an index of stabilized microtubules, and the level was reduced by Diaph3 expression. These results suggest that Diaph3 is required for cytokinesis only under high temperature conditions. Therefore, our study provides a new insight into the mechanisms by which regulatory factors of cell division function in a temperature-dependent manner.


Sign in / Sign up

Export Citation Format

Share Document