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Journal Article

Antioxidant and anti-proliferative activity of different solvent extracts of Casuarina equisetifolia needles

TL;DR: The results indicated that the needles of C. equisetifolia are rich sources of antioxidants and also contain potential anticancer agents.
Abstract: The context and purpose of the study: Plants that have been used for traditional medicines are very good sources of phyotochemicals There are many plants like Casuarina equisetifolia , which are still unexplored for their medicinal properties In the present study, we have elucidated the in vitro antioxidant and antiproliferative activity of different solvent extracts (both polar and non polar) of C equisetifolia needles Main findings: In vitro antioxidant activity of different solvent extracts of C equisetifolia needles was studied by analyzing the total polyphenols, flavonoids, total antioxidant capacity and free radical scavenging activity The polar solvent extracts showed significantly high amount total polyphenols, flavonoids, antioxidants and free radical scavenging activity compared to the non polar solvent extracts The cytotoxic and apoptosis inducing activity of the different solvent extracts were analyzed on MCF-7 cells by MTT assay, acridine orange/ethidium bromide staining, DAPI staining and caspase-3 release The polar solvent extracts did not show any growth inhibition in MCF-7 cancer cells But the non polar solvent extracts are very good in inducing cell death by inducing apoptosis which involves DNA fragmentation and release of caspase 3 Using silica gel fractionation and RP-HPLC analysis the active component present in non-polar solvent extracts was identified as ascorbic acid Brief summary and potential implications: Our results indicated that the needles of C equisetifolia are rich sources of antioxidants and also contain potential anticancer agents Detailed study on the mechanism of action of purified compound on inhibition of cancer cell growth may provide some potential anticancer molecule from natural source
Citations
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Journal ArticleDOI
TL;DR: The study showed that MECE is a potent wound healing agent through anti-inflammatory and antioxidant effects that enhanced the rate of wound contraction, re-epithelialization, and collagen deposition.
Abstract: Clausena excavata is a well-known plant used in folkloric medicine for the treatment of different ailments. This study aimed to determine the in vitro cytoxicity of its leaf solvent extracts as well as the in vivo wound healing and antioxidant activities of the methanolic extracts of C. excavata (MECE). HaCaT (keratocyte) and Vero cell lines were used for evaluation of the in vitro cytotoxic effects, while the in vivo wound healing and antioxidant activities were determined in skin wounds inflicted on rats. Twenty adult male Sprague-Dawley rats were divided into five groups of four animals each. Approximately 3.14 cm(2) excisional wound was inflicted on the nape of each rat following anesthesia. The treatment groups received topical application of MECE at 50 mg/mL (MECE-LD [low dose]), 100 mg/mL (MECE-MD [medium dose]), and 200 mg/mL (MECE-HD [high dose]), while the negative control group was treated with gum acacia in normal saline and the positive control group with intrasite gel. Wound contraction was evaluated on days 5, 10, and 15 after wound infliction, and tissue from wound area was collected at day 15 post-wound infliction for antioxidant enzyme evaluation and histopathological analyses. Generally, Vero cells were more resistant to the cytotoxic effects of the solvent extracts as compared with HaCaT cells. Chloroform (CH) and ethyl acetate (EA) extracts of C. excavata were toxic to HaCaT cells at 200 and 400 µg/mL, but the same concentrations showed higher (P<0.05) viability in Vero cells. There was significantly (P<0.01) greater wound contraction at days 10 and 15 post-wound infliction in all the treatment groups than in the control groups. Histopathologically, the MECE-HD-treated wound showed significantly (P<0.05) lesser inflammatory cell proliferation, degeneration, and distribution of granulation tissue than other groups. Similarly, the degree of collagen maturation, angiogenesis, and collagen distribution were significantly (P<0.05) lower in MECE-HD than in other groups. The MECE-HD, MECE-MD, and intrasite treatment groups showed a significantly (P<0.05) higher number of VEGF-positive and TGF-β1-positive cells in the skin wound than the control groups. The activities of superoxide dismutase and catalase were significantly (P<0.01) higher in the MECE-HD and intrasite treatment groups than in the other groups. Lipid peroxidase activity of the treated groups was significantly (P<0.01) lower than that in the control group. The study showed that MECE is a potent wound healing agent through anti-inflammatory and antioxidant effects that enhanced the rate of wound contraction, re-epithelialization, and collagen deposition. The effect of MECE is suggested to be due to its high polyphenolic compound content.

27 citations


Cites background or result from "Antioxidant and anti-proliferative ..."

  • ...This may indicate that nonpolar active principles are responsible for the antiproliferative activity of this plant as previously reported in other plants.(25,26) High Table 1 effect of Clausena excavata on percentage of wound healing in experimental rats...

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  • ...The antioxidant enzymes (SOD, CAT, and GPx) are known to overcome radicals and thus prevent the damage to cells caused by them.(25,30) This was shown in a previous study using Plagiochasma appendiculatum Lehm, where enhanced wound healing rate was attributed to decreased activity of LPO and increased activities of SOD and CAT....

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Journal ArticleDOI
TL;DR: This study is the first to reveal anti-E.
Abstract: Background. Clausena excavata Burum. f. has long been applied in ethnomedicine for the treatment of various disorders like rhinitis, headache, cough, wound healing, fever, and detoxification. This study is aimed at investigating the antibacterial activity against Enterococcus faecalis ATCC 49532 using AlamarBlue assay and atomic force microscopy (AFM) as well as the cytotoxicity, anticancer, and phytotoxicity of C. excavata. Method. Bacterial cell viability was performed by using microplate AlamarBlue assay. Atomic force microscopy was used to determine morphological changes in the surface of bacterial cells. Cytotoxicity and phytotoxicity were determined by brine shrimp lethality and Lemna minor bioassay. Caco-2 (colorectal adenocarcinoma) cell line was used for the evaluation of the anticancer effects. Result. Among the fractions tested, ethyl acetate (EA) fraction was found to be active with minimum inhibitory concentration (MIC) of 750 μg/mL against E. faecalis, but other fractions were found to be insensitive to bacterial growth. Microscopically, the EA fraction-treated bacteria showed highly damaged cells with their cytoplasmic content scattered all over. The value of the EA fraction against brine shrimp was more than 1000 μg/mL showing the nontoxic nature of this fraction. Chloroform (CH), EA, and methanol (MOH) fractions of C. excavata were highly herbicidal at the concentration of 1000 μg/mL. EA inhibited Caco-2 cell line with an of 20 μg/mL. Conclusions. This study is the first to reveal anti-E. faecalis property of EA fraction of C. excavata leaves, natural herbicidal, and anticancer agents thus highlight the potential compound present in its leaf which needs to be isolated and tested against multidrug-resistant E. faecalis.

9 citations


Cites background from "Antioxidant and anti-proliferative ..."

  • ...faecalis activity, as previously reported in other plants [50]....

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Dissertation
01 Feb 2015
TL;DR: In this paper, the presence of phytochemicals, antioxidant effect and antiproliferative activity of polygonum minus leaves extracts were investigated for detecting phytochemical constituents using various standard procedures.
Abstract: Cancer is one of the leading causes of death in the world, particularly in developing countries. Cancer prevention by dietary constituents has emerged as a novel approach to reduce the number of cancer incidence. The Polygonum minus leaves or commonly known as ‘Kesum’ in Malaysia, have been used as natural remedy in traditional medicine. The leaves of this plant have been reported to be high in antioxidants. Thus, this study was carried out to investigate the presence of phytochemicals, antioxidant effect and antiproliferative activity of P. minus leaves extracts. In the present study, ethanol and n-hexane extracts of P. minus leaves were examined for the presence of phytochemical constituents using various standard procedures. Folin-Ciocalteau’s method was used for the evaluation of total phenolic content of the extracts. The antioxidant activity of the extracts was measured using DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals scavenging activity assay. The antiproliferative activity of the extracts was examined against colon cancer cell line (HT 29) using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The ethanolic extract of P. minus showed the presence of more phytoconstituents than n-hexane extract. Ethanolic extract showed higher total phenolic content (137.07 ± 29.17 mg GAE/L) and strong antioxidant effect (IC50 = 63.1 µg/ml). The n-hexane extract exerted better antiproliferative activity against colon cancer cells (IC50 = 316 µg/ml). These data shows that ethanolic extract of P. minus leaves possesses antioxidant effect and n-hexane extract demonstrated antiproliferative activity against colon cancer cells.

3 citations

Journal Article
01 Jan 2007-Scopus
TL;DR: Ascorbic acid was found to reduce the proliferation of cells and induce apoptosis by the modulation of p53, p21, Bcl-2 and Bax and the anti-proliferative effects of AA against leukemic cells were shown.
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2 citations

References
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Journal ArticleDOI
09 Nov 2000-Nature
TL;DR: Evidence that the appropriate and inappropriate production of oxidants, together with the ability of organisms to respond to oxidative stress, is intricately connected to ageing and life span is reviewed.
Abstract: Living in an oxygenated environment has required the evolution of effective cellular strategies to detect and detoxify metabolites of molecular oxygen known as reactive oxygen species. Here we review evidence that the appropriate and inappropriate production of oxidants, together with the ability of organisms to respond to oxidative stress, is intricately connected to ageing and life span.

8,665 citations

Journal ArticleDOI
TL;DR: The phosphomolybdenum method is routinely applied in the laboratory to evaluate the total antioxidant capacity of plant extracts and to determine vitamin E in a variety of grains and seeds, including corn and soybean.

4,644 citations


"Antioxidant and anti-proliferative ..." refers methods in this paper

  • ...This assay is based on the reduction of Mo (VI) to Mo (V) by the sample and the subsequent formation of a green phosphate/Mo (V) complex at acidic pH [9] using ascorbic acid as standard....

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Journal Article
TL;DR: In this article, a 40-sample/hour procedure was adapted from the Singleton-Rossi method of analysis for total phenols in wine and other plant extracts, and compared with small-volume manual and semi-automated versions of this analysis.
Abstract: A fully automated-continuous flow 40-sample/ hour procedure was adapted from the Singleton-Rossi method of analysis for total phenols in wine and other plant extracts. It was compared with small-volume manual and semiautomated versions of this analysis. The agreement in mg of gallic acid equivalent phenol (GAE) per liter among a series of dry wines was excellent by all three procedures. The coefficients of variation in replicate analyses averaged 5.8% for the manual, 6.2% for the semi-automated and 2.2% for the automated procedure. This greater reproducibility, plus savings of about 70% in labor and up to 40% in reagents, makes the automated procedure attractive for laboratories doing enough total phenol analyses to recoup the cost of the automating equipment. For continuous flow, color development with the Folin-Ciocalteu reagent in alkaline solution must be hastened by heating compared to slower room temperature development for the manual methods. Heating of sugar-containing samples in the alkaline solution gives interference presumably from endiol formation. Examples are given of corrections which were used successfully to estimate the true phenol content of sweet wines.

3,527 citations


"Antioxidant and anti-proliferative ..." refers methods in this paper

  • ...Total phenolic content of each extract was determined by the Folin– Ciocalteu micro-method [7] using gallic acid as standard....

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Journal Article
TL;DR: The new XTT reagent provides for a simplified, in vitro cell growth assay with possible applicability to a variety of problems in cellular pharmacology and biology, but still shares many of the limitations and potential pitfalls of MTT or other tetrazolium-based assays.
Abstract: We have previously described the application of an automated microculture tetrazolium assay (MTA) involving dimethyl sulfoxide solubilization of cellular-generated 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-formazan to the in vitro assessment of drug effects on cell growth (M.C. Alley et al., Proc. Am. Assoc. Cancer Res., 27:389, 1986; M.C. Alley et al., Cancer Res. 48:589-601, 1988). There are several inherent disadvantages of this assay, including the safety hazard of personnel exposure to large quantities of dimethyl sulfoxide, the deleterious effects of this solvent on laboratory equipment, and the inefficient metabolism of MTT by some human cell lines. Recognition of these limitations prompted development of possible alternative MTAs utilizing a different tetrazolium reagent, 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl] -2H- tetrazolium hydroxide (XTT), which is metabolically reduced in viable cells to a water-soluble formazan product. This reagent allows direct absorbance readings, therefore eliminating a solubilization step and shortening the microculture growth assay procedure. Most human tumor cell lines examined metabolized XTT less efficiently than MTT; however, the addition of phenazine methosulfate (PMS) markedly enhanced cellular reduction of XTT. In the presence of PMS, the XTT reagent yielded usable absorbance values for growth and drug sensitivity evaluations with a variety of cell lines. Depending on the metabolic reductive capacity of a given cell line, the optimal conditions for a 4-h XTT incubation assay were 50 micrograms of XTT and 0.15 to 0.4 microgram of PMS per well. Drug profiles obtained with representative human tumor cell lines for several standard compounds utilizing the XTT-PMS methodology were similar to the profiles obtained with MTT. Addition of PMS appeared to have little effect on the metabolism of MTT. The new XTT reagent thus provides for a simplified, in vitro cell growth assay with possible applicability to a variety of problems in cellular pharmacology and biology. However, the MTA using the XTT reagent still shares many of the limitations and potential pitfalls of MTT or other tetrazolium-based assays.

2,380 citations


"Antioxidant and anti-proliferative ..." refers background in this paper

  • ...equisetifolia on human breast cancer MCF-7 cell’s viability [12]....

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Journal ArticleDOI
01 Jan 1998-Nature
TL;DR: The results indicate that activation of CAD downstream of the caspase cascade is responsible for internucleosomal DNA degradation during apoptosis, and that ICAD works as an inhibitor of this process.
Abstract: Various molecules such as cytokines and anticancer drugs, as well as factor deprivation, rapidly induce apoptosis (programmed cell death), which is morphologically characterized by cell shrinkage and the blebbing of plasma membranes and by nuclear condensation. Caspases, particularly caspase 3, are proteases that are activated during apoptosis and which cleave substrates such as poly(ADP-ribose) polymerase, actin, fodrin, and lamin. Apoptosis is also accompanied by the internucleosomal degradation of chromosomal DNA. In the accompanying Article, we have identified and molecularly cloned a caspase-activated deoxyribonuclease (CAD) and its inhibitor (ICAD). Here we show that caspase 3 cleaves ICAD and inactivates its CAD-inhibitory effect. We identified two caspase-3 cleavage sites in ICAD by site-directed mutagenesis. When human Jurkat cells were transformed with ICAD-expressing plasmid, occupation of the receptor Fas, which normally triggers apoptosis, did not result in DNA degradation. The ICAD transformants were also resistant to staurosporine-induced DNA degradation, although staurosporine still killed the cells by activating caspase. Our results indicate that activation of CAD downstream of the caspase cascade is responsible for internucleosomal DNA degradation during apoptosis, and that ICAD works as an inhibitor of this process.

1,612 citations

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