Morin Inhibits Proliferation of SW480 Colorectal Cancer Cells by Inducing Apoptosis Mediated by Reactive Oxygen Species Formation and Uncoupling of Warburg Effect

The study under investigation focuses on in vitro antiproliferative efficacy of the flavonoid morin and the mechanisms by which it inhibits the growth of colon cancer using SW480 colon cancer cells with emphasis on Warburg effect. It was found that the cell proliferation was significantly inhibited by morin in a dose and time dependent manner. Morin induced apoptosis that was correlated with increased levels of reactive oxygen species formation and loss of mitochondrial membrane potential of the cells. In addition, an increase in cleaved PARP, cleaved caspase 3, cleaved caspase 8, cleaved caspase 9 and Bax as well as a decrease in Bcl 2 was observed, indicating morin is inducing both intrinsic as well as extrinsic pathway of apoptosis. This was further confirmed by using downstream caspase 3 inhibitor which indicated that caspase 3 inhibition reduces morin induced cell death. Moreover, the impact of morin on over all energy status when determined in terms of total cellular ATP level showed a decline with low level of glucose uptake and Glut1 expression. The results indicate that morin exerts antiproliferative activity by inducing apoptosis and by reducing Warburg effect in the evaluated cell lines and provide preliminary evidence for its anticancer activity

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Not AvailableA blend of plant proteins (soybean meal, groundnut oil cake, sunflower oil cake, wheat gluten meal and corn gluten meal at 4:2:1:6.5:6.5) was used to substitute fishmeal in the diet of Asian seabass Lates calcarifer (Bloch, 1790). Five iso-nitrogenous (401.13-407.33 g kg-1) diets were prepared by substituting fishmeal at 0 (control), 25, 50, 75 and 100% (FM-R0, FM-R25, FM-R50, FM-R75 and FM-R100, respectively) using the above mix. Average final weight of the fish increased (p<0.05) by three-folds (20.77-29.99 g) after 45 days compared to their initial weight (6.09±0.25 g) in all the treatments, however, significant (p<0.05) variation was observed between the dietary treatments (p<0.002). The results indicated that 50% of dietary fishmeal can be substituted and beyond this level resulted in significant reduction of growth performance. Broken-line analysis indicated optimal fishmeal substitution as 46.7%. The highest activity of pepsin, trypsin, chymotrypsin, carboxypeptidases-A and B was observed in fish fed with diets FM-R0, FM-R25 and FM-R50. Dietary change significantly (p<0.05) influenced the carcass lipid composition of Asian seabass (p<0.001). Fatty acids like C20:4, C20:5 and C22:6 were significantly (p<0.05) low in fish carcass fed with higher levels of plant proteins (FM-R75 and FM-R100). The results concluded that dietary fishmeal level can be partially substituted using a blend of plant proteins in the diet of Asian seabass juveniles.Not Availabl

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Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake.

Enhanced oxidative stress contributes to pathological changes in diabetes and its complications. Thus, strategies to reduce oxidative stress may alleviate these pathogenic processes. Herein, we have investigated Naringin mediated regulation of glutathione (GSH) & intracellular free radical levels and modulation of glucose uptake under oxidative stress in L6 cell lines. The results from the study demonstrated a marked decrease in glutathione with a subsequent increase in free radical levels, which was reversed by the pretreatment of Naringin. We also observed that the increased malondialdehyde level, the marker of lipid peroxidation on induction of oxidative stress was retrieved on Naringin pretreatment. Addition of Naringin (100 μM) showed approximately 40% reduction in protein glycation in vitro. Furthermore, we observed a twofold increase in uptake of fluorescent labeled glucose namely 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) on Naringin treatment in differentiated L6 myoblast. The increased uptake of 2-NBDG by L6 myotubes may be attributed due to the enhanced translocation of GLUT4. Our results demonstrate that Naringin activate GSH synthesis through a novel antioxidant defense mechanism against excessive Reactive Oxygen Species (ROS) production, contributing to the prevention of oxidative damage in addition to its effect on glycemic control

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GLUT4 upregulation on Naringin pretreatment.

<p>Immunofluorescence assay visualized upregulation of GLUT4 in differentiated L6 myoblast. High resolution confocal images (40X) of Untreated L6 myotubes, L6 myotubes treated with TBHP, Rosiglitazone (100 nM) and Naringin (100 μM, 24 h). Scale bar corresponds to 10μM. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups were determined by using one way ANOVA followed by Duncan’s multiple range test. <b>*</b> P≤0.05 versus control; #P≤0.05 versus TBHP.</p

Intracellular ROS production and Fluorescence intensity analysis in L6 myoblast.

<p>(A) Fluorescence images (20 X magnifications) of untreated cell (i); Figures (ii), (iii) & (iv) represents cells induced with TBHP at 1, 10 & 100 μM; (v) Represents the relative fluorescence intensity analysis by BD Image Data Explorer software. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. *P≤0.05 versus control. (B) Figure (i), (ii), (iii), (iv) & (v) represents fluorescence images of untreated cells, cells induced with TBHP (100 μM) and cells pretreated with Naringin (1, 10 & 100 μM) for 3h respectively; (vi) Represents the relative fluorescence intensity analysis by BD Image Data Explorer software. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. *P≤0.05 versus control; #P≤0.05 versus TBHP.(C) Figure (i), (ii), (iii), (iv) & (v) represents fluorescence images of untreated cells, cells induced with TBHP (100 μM) and cells pretreated with Naringin (1, 10 & 100 μM) for 24h respectively; (vi) Represents the relative fluorescence intensity analysis by BD Image Data Explorer software. Scale bar corresponds to 87 μM. TBHP: Tertiary butyl hydrogen peroxide; Na1+TBHP, Na2+TBHP & Na3+TBHP represents relative fluorescence intensity analysis of cells pretreated with Naringin (1, 10 & 100 μM) followed by induction of TBHP; Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.* P≤0.05 versus control; #P≤0.05 versus TBHP.</p

Fluorescence analysis of 2-NBDG uptake by flow cytometry.

<p>FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.<b>*</b> P≤0.05 versus control.</p