Antioxidant And Anti-Inflammatory Activities Of Leaves, Calli And Cell Suspension Of Putat (Barringtonia Racemosa)

The medicinal plant of Barringtonia racemosa (Lecythidaceae family) has been used widely in traditional medicine for anti-inflammation and anticancer in Malaysia. The present investigation was carried out to study anti-oxidant and anti-inflammatory effects of leaves, callus, cell suspension and in vitro regenerated shoots and roots of B. racemosa. The results showed that different crude extracts of fully expanded leaf extracts of B. racemosa have a very strong nitric oxide (NO) inhibitory and antioxidant activities. In the Griess assay, non polar extracts such as chloroform and hexane extracts were found to be strong inhibitors of NO at different concentrations (25, 50, 100 and 200 μg/ml) in comparison with polar extract (ethanol extract).Calli were aseptically obtained by placing surface sterilized leaf explants on Woody Plant Medium (WPM) supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D). On the shoot induction medium, the callus induced on the WPM medium containing 2 mg/L (w/v) KIN+0.2 mg/l (w/v) IBA and 2 mg/L (w/v) of KIN + 0.4 mg/L(w/v) of NAA was the most effective, providing high shoot regeneration frequency of 85.6 and 76.5 %, respectively. In addition, the highest number of shoots produced was 8.2 and 6.3 shoots per explant respectively in the medium containing the mentioned plant growth regulators. The rooting percentage and number of roots per shoot which achieved on WPM medium supplemented with 3g/L (w/v) of activated charcoal and 0.8 mg/L (w/v) of IBA were 62 and 5.6 %, respectively. 96 % of the in vitro rooted plantlets with well developed shoots and roots were survived when transferred to soil. Results obtained from this study revealed that B.racemosa is one of the important sources of lycopene. Lycopene has long been recognized as important antioxidants both in vivo and in vitro. Lycopene level was detected at a range of 0.02 to 4.14 mg/g dry weight in in vitro regenerated shoots and roots respectively. Lycopene level was also successfully detected in the callus (0.34 to 2.12 mg/g dry weight) and cell suspension cultures (0.18 to 0.68 mg/g dry weight) under dark and light conditions and the amount was lower than that produced in the intact plant tissues. However, manipulating the physical conditions, feeding of precursor and elicitation managed to increase the lycopene content in cultured tissues. Studies on the effects of the medium composition show that fully strength of the basal Woody Plant Medium and B5 containing 3% (w/v) of sucrose increased the lycopene content in both callus and cell suspension cultures. The precursor-feeding studies revealed that concentrations of 3 mg/L (w/v) of isopentenyl pyrophosphate and 2 to 4 mg/L (w/v) of Mevalonate were preferred for lycopene production. The elicitor studies exhibited that the different elicitors showed distinctive effects on lycopene production. Nevertheless, casein hydrolysate at 10 and 15 mg/l (w/v) was found to be the best in increasing the lycopene production in callus and cell suspension cultures. The study further concluded that there was correlation between anti-oxidant and anti-inflammatory activities and lycopene content in callus, cell suspension and in vitro regenerated organs of B.racemosa

Otimização de culturas de suspensões de calos e células de Barringtonia racemosa (família Lecythidaceae) para produção de licopeno

Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC) and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS), Woody Plant Medium (WPM) and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D). Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume) of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC) method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.O licopeno está presente numa série de frutas frescas e hortaliças principalmente na folhas de Barringtonia racemosa. A extração tradicional do licopeno tem sido empregada no lugar da fácil técnica de propagação como o processo de cultura de células de explantes de folhas. É nossa intenção demonstrar como o licopeno pode ser extraído através de cultura de tecido sob luz (iluminação com lâmpadas fluorescentes brancas de 8.200 lux, 16 h por dia a 25º C) e escuro. Explantes de folhas de Barringtonia racemosa foram cultivados em meio modificado de Murashige e Skoog (MS) para plantas lenhosas e meio B5, suplementado com diferentes concentrações de ácido 2,4-Diclorofenoxiacético (2,4-D). Condições ótimas para indução e manutenção de calos sob luz e escuro foram investigadas e avaliados o crescimento e acumulo de licopeno. Entre meios com diferentes concentrações de 2,4 -D, calos friáveis de crescimento rápido tiveram início em três semanas após serem cultivados em meio basal WPM suplementado com 2.0 mg L-1 (peso por volume) de 2,4-D enquanto indução de calos em explantes cultivados em todos os outros meios começaram somente após cinco semanas. Calos foram subrepicados a cada 15 dias. Calos amarelo-pálido e verdes desenvolvidos respectivamente sob condições escura e de luz foram então selecionados para avaliação do teor de licopeno. Um método aperfeiçoado de cromatografia líquida de alto desempenho foi usado para a determinação química seletiva do teor de licopeno. A produção de licopeno induzida sob luz e também o nível máximo de licopeno incubado em luz foi mais alto do que aqueles incubados no escuro. As melhores taxas de crescimento de calo e suspensões de células foram obtidas respectivamente em meio WPM e B5. A produção de licopeno dependeu do crescimento como demonstrado pela análise do crescimento e teor de licopeno de ambos calos e cultura de células em suspensão

Antibacterial and antibiofilm activities of Prangos acaulis Bornm. extract against Streptococcus mutans: an in silico and in vitro study

Introduction: Streptococcus mutans is a principal pathogenic agent in biofilm formation on the teeth surfaces and subsequently development of dental caries and plaque. Therefore, currently introducing novel anti-bacterial and anti-biofilm agents, especially plant based materials are highly regarded. This study was planned to investigate in silico and in vitro antibacterial activities of Prangos acaulis extracts against S. mutans in single and biofilm forms and their mutagenicity in Ames test. Methods: The anti-bacterial and anti-biofilm effects of methanol extracts from various parts of P. acaulis were evaluated using disk diffusion and microtiter assay. Moreover, the potential mutagenicity of the extracts was investigated using Ames test. In addition, dominant constitutes of P. acaulis that reported in previous studies were subjected to an in silico analysis. The ability of selected phytochemicals to inhibit the glucosyltransferase was evaluated using molecular docking method. Results: All tested extracts especially root extract had significant antibacterial activity against the single form of S. mutans and inhibited biofilm formation without any mutagenic activity. The results also confirmed that three compounds consisting of ar-curcumene, d-limonene and alpha-pinene had strong and appropriate interactions to glucosyltransferase. Conclusion: This study indicated that P. acaulis has potent antibacterial and biofilm inhibition activity against S. mutans and can be good candidate for in vitro and in vivo studies with the aim of introducing novel inhibitors of dental caries developmen

Anti-HIV-1 activity of eight monofloral Iranian honey types.

Monofloral Iranian honeys from eight floral sources were analyzed to determine their anti-HIV-1 activities as well as their effects on lymphocyte proliferation. The Peripheral Blood Mononuclear Cells (PBMCs) used in this study were prepared from five healthy volunteers who were seronegative for HIV, HCV, HBV and TB. The anti-HIV-1 activity of eight different honeys was performed by quantitative polymerase chain reaction (PCR) assay and high pure viral nucleic acid kit. The results demonstrated that monofloral honeys from Petro selinum sativum, Nigella sativa, Citrus sinensis, Zataria multiflora, Citrus aurantium and Zizyphus mauritiana flowers had potent anti-HIV-1 activity with half maximal effective concentration (EC50) values of 37.5, 88, 70, 88, 105 and 5 µg/ml respectively. However, monofloral Iranian honeys from Astragalus gummifer and Chamaemelum nobile flowers had weak anti-HIV-1 activity. The frequency and intensity of CD4 expression on PBMCs increased in the presence of all honey types. CD19 marker were also increased after the treatment with monofloral honeys from Z. multiflora and N. sativa. The anti-HIV-1 agent in monofloral honeys from P. sativum, N. sativa, Z. multiflora and Z. mauritiana flowers was detected by spectroscopic analysis as methylglyoxal. Time of drug addition studies demonstrated that the inhibitory effect of methylglyoxal is higher on the late stage of HIV-1 infection. The result demonstrated that methylglyoxal isolated from monofloral honey types is a good candidate for preclinical evaluation of anti-HIV-1 therapies

Evaluation of in vitro anticancer activity of Ocimum basilicum, Alhagi maurorum, Calendula officinalis and their parasite Cuscuta campestris.

The present investigation was carried out to study the relationship between presence of cytotoxic compounds in Ocimum basilicum, Alhagi maurorum, Calendula officinalis and their parasite Cuscuta campestris. The cytotoxic activity of the pure compounds was performed by MTT assay against breast cancer cell lines (MCF-7 and MDA-MB-231) and normal breast cell line (MCF 10A). The induction of apoptosis was measured by the expression levels of p53, bcl-2, bax and caspase-3 genes using quantitative Real Time PCR. Three active fractions were detected by nuclear magnetic resonance as lutein, lupeol and eugenol, respectively, in C. officinalis, A. maurorum and O. basilicum. These compounds and their epoxidized forms were also detected in their parasite C. campestris. The cytotoxic activity of lutein epoxide, lupeol epoxide and eugenol epoxide was significantly more than lutein, lupeol and eugenol. The mRNA expression level of p53, caspase-3 and bax genes were increased in both cancer cells treated with all pure compounds. However, bcl-2 gene expression decreased in treated breast cancer cells. In conclusion, all the data indicated that the epoxide forms of lupeol, lutein and eugenol are potential drug candidates for inducing apoptosis in human breast cancer cells

Cytotoxic activity of lutein epoxide, lupeol epoxide, eugenol epoxide, lutein, lupeol and eugenol against MCF7, MDA-MB231 and MCF 10A cell lines.

<p>Cytotoxic activity of lutein epoxide, lupeol epoxide, eugenol epoxide, lutein, lupeol and eugenol against MCF7, MDA-MB231 and MCF 10A cell lines.</p

Time dependency effects of p53 and bcl-2 mRNA levels in human breast cancer cell line, MCF7, incubated with <i>lutein, lupeol, eugenol, lutein epoxide, lupeol epoxide, eugenol</i> epoxide at 1/4 of CC50 values for 6 h and 12 h incubation.

<p>gapdh was used as an endogenous control gene. The stars indicate that the data are significantly different (p<0.05) from the untreated control.</p

Time of addition effect of methylglyoxal (□) and AZT (•) at concentration of 5 µg/ml on HIV-1 replication.

<p>The extracts were added 0, 2, 4, 6, 8, 10, 16, 24, 32, 48 hours after virus infection. Each value is the result of mean ± SD of three independent experiments. The stars indicate that the data are significantly different (p<0.05) from the untreated control.</p

Western blot analysis of MDA-MB-231 and MCF7 treated with lutein (A), lupeol (B), eugenol (C), lutein epoxide (D), lupeol epoxide (E) and eugenol epoxide (F).

<p>Western blot analysis was performed with monoclonal antibodies to human bcl-2, p53 and bax. β-actin was used as loading control.</p