Phytochemical investigations and antibacterial activity of Salacia oblonga Wall ethanolic extract

Abstract Salacia oblonga Wall, a medicinally important plant, belonging to the family Celastraceae, is a large woody climber distributed in southern India and Sri Lanka. In the present investigation, ethanol extracts of S. oblonga were prepared from aerial and root parts of the plant in the presence and absence of HCl and antibacterial activity was tested. Both aerial and root extracts exhibited pronounced activity against human pathogens. The MIC and MBC values ranged from 0.078-1.25 mg/ml and 0.156 -2.50 mg/ml, respectively. GC-MS profile of aerial and root extracts displayed the presence of 11 and 6 compounds. The present investigation demonstrated that ethanolic extracts of S. oblonga have potential antibacterial activity against human pathogens and could serve as a source for the development of new age antimicrobials

EFFECT OF SALACIA OBLONGA ROOT EXTRACT AGAINST CLINICAL ISOLATES STAPHYLOCOCCUS AUREUS

Objective: Salacia oblonga Wall. is an important medicinal plant belonging to the family Celastraceae. The study reports the effect of S. oblonga root extracts against clinical isolate Staphylococcus aureus Methods: Antibacterial activity was evaluated by agar diffusion method and assay for minimum inhibitory concentration (MIC) of extract. Further, the effect of S. oblonga extract determined by DNA fragmentation and respiratory dehydrogenase enzyme activity assays. Results: S. oblonga ethyl acetate root extract was evaluated for antibacterial activity towards clinical isolate S. aureus. Bacterial growth was determined in treated and control cells. Extract displayed good growth inhibition and MIC of the extract was 80 μg/ml. DNA fragmentation assay was carried out, this result has shown that treated bacterial cell has DNA damage compared to the control cell. Further, respiratory dehydrogenase enzyme activity was determined. In the treated cells, enzyme activity was low compared to the control cells. Conclusion: Salacia oblonga root extract inhibiting the growth of S. aureus by different modes of action

Extracts of Strawberry Fruits Induce Intrinsic Pathway of Apoptosis in Breast Cancer Cells and Inhibits Tumor Progression in Mice

Background: The consumption of berry fruits, including strawberries, has been suggested to have beneficial effects against oxidative stress mediated diseases. Berries contain multiple phenolic compounds and secondary metabolites that contribute to their biological properties. Methodology/Principal Findings: Current study investigates the anticancer activity of the methanolic extract of strawberry (MESB) fruits in leukaemia (CEM) and breast cancer (T47D) cell lines ex vivo, and its cancer therapeutic and chemopreventive potential in mice models. Results of MTT, trypan blue and LDH assays suggested that MESB can induce cytotoxicity in cancer cells, irrespective of origin, in a concentration-and time-dependent manner. Treatment of mice bearing breast adenocarcinoma with MESB blocked the proliferation of tumor cells in a time-dependent manner and resulted in extended life span. Histological and immunohistochemical studies suggest that MESB treatment affected tumor cell proliferation by activating apoptosis and did not result in any side effects. Finally, we show that MESB can induce intrinsic pathway of apoptosis by activating p73 in breast cancer cells, when tumor suppressor gene p53 is mutated. Conclusions/Significance: The present study reveals that strawberry fruits possess both cancer preventive and therapeutic values and we discuss the mechanism by which it is achieved

Evaluation of side effects of MESB in mice.

<p>Mice were orally fed with MESB (2 g/kg. b. wt) for 10 days. <b>A.</b> Data showing average body weight changes in the control (n = 8) and MESB treated mice (n = 8). In all the cases, error bars are indicated. <b>B.</b> Hematological and serum profile of mice following oral feeding with MESB at day 10 (n = 8 for both control and treated). Values are indicated in mean ± SEM.</p

Proposed model for mechanism of MESB induced cytotoxicity.

<p>MESB treatment resulted in activation of intrinsic pathway of apoptosis. This is mediated through activation of p73. This activation leads to changes in the level of mitochondrial apoptotic protein, BAX. This may result in the imbalance of proapoptotic/antiapoptotic proteins. The activation of BAX, further leads to cleavage of MCL-1 and release of CYTOCHROME C, which along with APAF1 helps in cleavage of CASPASE 9. Cleaved CASPASE 9 activates CASPASE 3 which further initiates PARP1 cleavage and cell death.</p

Expression of apoptotic proteins in T47D cells following MESB treatment.

<p>Whole cell extracts (<b>A-C</b>) and cyotosolic extracts (<b>D</b>) were prepared from T47D cells following treatment with MESB (0, 0.1, 0.4, 0.7 mg/ml for 48 h). Western blotting studies were performed using primary antibodies against <b>(A)</b> MCL-1, BCL-xL, BAX and BID, <b>(B)</b> p53, MDM2, p73 and PARP1, <b>(C)</b> SMAC/DIABLO, CYTOCHROME C, APAF1, CASPASE 3 and CASPASE 9, (D) SMAC/DIABLO and CYTOCHROME C. In panels A-C, TUBULIN was used as an internal loading control, while in D, ACTIN was used.</p

Histopathology of the tumor tissues and liver of mice following MESB treatment.

<p>Histopathalogical sections of thigh and liver of a tumor bearing mouse with and without treatment with MESB after 30^th day A(a–f) and C(a–f) and 45^th day B(a–f) and D(a–f) of development of tumor. Magnification shown are 10x (a, c and e in all panels) and 40x (b, d and f in all panels).</p

Assessment of MESB induced cytotoxicity in leukemic (CEM) and breast cancer (T47D) cell lines.

<p>CEM and T47D cells were treated with increasing concentrations of MESB (0.1, 0.2, 0.5 and 1 mg/ml) and cells were harvested after 48 or 72 h of treatment and subjected to trypan blue, MTT and LDH assays. <b>A.</b> Determination of cell viability by trypan blue assay in CEM cells. <b>B.</b> Evaluation of cell proliferation by MTT assay in CEM cells. <b>C.</b> Bar diagram showing release of lactate dehydrogenase following MESB treatment in CEM cells. <b>D.</b> Assessment of cell viability using trypan blue assay in T47D cells. <b>E.</b> Determination of cell proliferation by MTT assay in T47D cells after MESB treatment. <b>F.</b> LDH assay showing release of lactate dehydrogenase in T47D cells following addition of MESB. In each panel, error bars were calculated based on results obtained from minimum of three independent experiments. In all panels, *p<0.05, **p<0.005, and ***p<0.0005.</p

Effect of MESB on progression of tumor in mice.

<p>Solid tumor was induced in Swiss albino mice by injecting breast adenocarcinoma cells. Following tumor development, mice were orally treated with MESB (2 g/kg body weight) until 45^th day. <b>A.</b> Effect of MESB on tumor progression at different time points. Data shown is derived from three independent experiments containing 10 animals each. Error bars indicate standard deviation. <b>B.</b> Kaplan-Meier survival curves of mice treated with MESB. <b>C.</b> The chemopreventive effect of MESB on EAC induced mice. The experimental mice were orally fed with MESB for 20 days prior to the EAC injection and were compared with control group, which did not receive pretreatment by MESB. <b>D.</b> Kaplan-Meier survival curves of mice pretreated with MESB.</p