Alteration of Bax-to-Bcl-2 ratio modulates the anticancer activity of methanolic extract of Commelina benghalensis (Commelinaceae) in Jurkat T cells

Stem extracts of Commelina benghalensis (Linn.), although not extensively documented, are frequently used in traditional medicine for the treatment of ailments such as skin malformations and outgrowths. Accordingly, the study was aimed to investigate possible molecular mechanisms that are associated with the potential anti-carcinogenic property of this agrofield weed. Jurkat T cells were exposed to different concentrations (0-600 ug/ml) of the crude methanolic extract of C. benghalensis to evaluate their growth inhibitory and apoptosis inducing effects. The extract elicited a dose- and time-dependent inhibition of cell proliferation, followed by a concomitant decrease in cell viability. The observed cytotoxicity was linked to the induction of apoptosis as determined by morphological and biochemical features known to be associated with the advent of apoptosis. Real time quantitative RT-PCR and Western blot analyses of Bax, Bcl-2 and p53 exhibited aberrant expression profiles of these genes under various treatment conditions. Taken together, the data suggest that the crude methanolic extract of C. benghalensis contains bioactive compounds that may be beneficial in the treatment of malignant growths, and that this apparent antineoplastic activity is a consequence of dysregulated expression of apoptosis-responsive genes. These observations could provide a credible scientific justification upon which the ethnopharmacological utilisation of C. benghalensis is founded

Antioxidant, Antiglycation, and Hypoglycaemic Effect of Seriphium plumosum Crude Plant Extracts

Diabetes is a severely debilitating metabolic disorder characterised by chronic hyperglycaemia. Traditional medicinal plants provide an important avenue for the development of novel antidiabetic agents. The antidiabetic potential of the methanol, acetone, and hexane extracts of S. plumosum was assessed using different parameters. These included secondary metabolite quantification, hypoglycaemic, cytotoxic effects, and GLUT4 translocation augmentation on C2C12 cells. The methanol extract contained the highest amount of total phenolic and flavonoid compounds and showed enhanced antioxidant activity. The methanol extracts had the best DPPH scavenging (EC50 = 0.72 mg/ml) and ferric reducing powers (EC50 = 2.31 mg/ml). The hexane extract resulted in the highest glucose uptake activity of 35, 77% with respect to all other treatments after a 6-hour exposure period. Immunocytochemistry technique further revealed that the increased glucose utilisation may be due to increased membrane fused GLUT4 molecules in C2C12 cells. The hexane extract was also shown to upregulate the phosphorylation of p70 S6 kinase and Akt1/2. The study highlights a probable insulin-mimetic activity of the hexane extract via the augmentation of Akt1/2 phosphorylation which is involved in the GLUT4 translocation pathway. Furthermore, the study represents the first report on the cytotoxic effect, GLUT4 translocation, and glucose uptake potential of S. plumosum

The Effect of Lithium on Inflammation-Associated Genes in Lipopolysaccharide-Activated Raw 264.7 Macrophages

Lithium remains the preferred Food and Drug Administration- (FDA-) approved psychiatric drug for treatment of bipolar disorders since its medical establishment more than half a century ago. Recent studies revealed a promising role for lithium in the regulation of inflammation, oxidative stress, and neurodegeneration albeit unclear about its exact mode of action. Thus, the intention of this study is to delineate the regulatory mechanisms of lithium on oxidative stress in lipopolysaccharide- (LPS-) activated macrophages by evaluating its effects on nuclear factor-κB (NF-κB) activity and mRNA expression of multiple oxidative stress-related NF-κB genes. Raw 264.7 macrophages were treated with up to 10 mM lithium, and no change in cell proliferation, viability, growth, and cell adhesion was observed in real time. Pretreatment with low doses of lithium was shown to reduce nitric oxide (NO) production in LPS-activated macrophages. A reduced internal H2DCFDA fluorescence intensity, indicative of reduced reactive oxygen species (ROS) production, was observed in LPS-activated Raw 264.7 macrophages treated with lithium. Lithium has been shown to lower the production of the chemokine RANTES; furthermore, this inhibitory action of lithium has been suggested to be independent of glycogen synthase kinase-3 β (GSK3β) activity. It is shown here that lithium modulates the expression of several inflammatory genes including IκB-α, TRAF3, Tollip, and NF-κB1/p50 which are regulators of the NF-κB pathway. Moreover, lithium inhibits NF-κB activity by lowering nuclear translocation of NF-κB in LPS-activated macrophages. This is the first study to associate Tollip, Traf-3, and IκB-α mRNA expression with lithium effect on NF-κB activity in LPS-activated Raw 264.7 macrophages. Although these effects were obtained using extratherapeutic concentrations of lithium, results of this study provide useful information towards understanding the mode of action of lithium. This study associates lithium with reduced oxidative stress in LPS-activated Raw 264.7 macrophages and further suggests candidate molecular targets for the regulation of oxidative stress-related diseases using lithium beyond bipolar disorders

Defatting of acetone leaf extract of Acacia karroo (Hayne) enhances its hypoglycaemic potential

Abstract Background Conventional drugs used to treat diabetes are too expensive, toxic and rarely available to rural communities. This study was aimed at investigating the phytochemical differences and hypoglycaemic effects (α-amylase enzyme inhibition, glucose uptake, GLUT4 translocation and phosphorylation of MAPKs) of non-defatted and defatted acetone leaf extract of Acacia karroo. Methods Qualitative phytochemical analyses of extracts were determined using standard chemical tests and total phenolic contents using the Folin-Ciocalteu reagent method. Presence of antioxidant constituents was determined using DPPH scavenging and ferric reducing power assays. Alpha amylase enzyme inhibitory potential was determined chromogenically and cytotoxicity of the extracts on C2C12 muscle and 3T3-L1 cells using the MTT assay. Glucose uptake by the cells was determined colorimetrically and the most active extract was evaluated for its ability to translocate GLUT4 and MAPKs phosphorylation potential using immunofluorescence microscopy and dot blot analysis, respectively. Results Phenols, flavonoids, tannins, saponins and cardiac glycosides were detected in both extracts. Defatting of the plant material resulted in low amounts of phenols (0.432 ± 0.014 TAE/mg), DPPH scavenging activity (EC50 0.40 ± 0.012 mg/ml), low toxicity and high ferric reducing power (EC50 1.13 ± 0.017 mg/ml), α-amylase enzyme inhibition (IC50 30.2 ± 3.037 μg/ml) and glucose uptake by both cells. The defatted extract showed an increase in GLUT4 translocation (at 25 μg/ml) with decrease in Akt expression while in combination with insulin showed a decrease in GLUT4 translocation. A finding, that is implicative that the effect of the extract on GLUT4 translocation in C2C12 cells was not Akt dependent. The defatted extract in the absence and presence of insulin show varying phosphorylation levels of CREB, p38, GSK-3 and ERK2 which are important in cell survival and metabolism. Conclusion This study represents the first report on the hypoglycemic potential of A. karroo and presence of compounds that can be exploited in the search for therapeutics with antidiabetic effect

Potential Antiglycation and Hypoglycaemic Effects of Toona ciliata M. Roem. and Schkuhria pinnata Lam. Thell. Crude Extracts in Differentiated C2C12 Cells

Medicinal plants have been identified as a feasible avenue for the development of new potent antidiabetic agents. The phytoconstituent compositions of different Toona ciliata and Schkuhria pinnata extracts were determined and quantified using standard chemical methods after exhaustive extraction. Thereafter, their antioxidant and antiglycation potentials were spectrophotometrically determined. The cytotoxicity profiles of the extracts on C2C12 cells were determined using the MTT assay. Toona ciliata methanol extract resulted in the highest percentage yield (20.83%) and high total phenols and flavonoids content in the methanol and acetone extracts compared to S. pinnata extracts. The acetone extract of T. ciliata showed good activity in the DPPH scavenging and FRAP assays with EC50 values of 1.90 mg/ml and 5.26 mg/ml, respectively. Arbutin’s antiglycation ability was outperformed by treatments with the methanol, acetone, and hexane extract of T. ciliata which resulted in 2.49%, 2.79%, and 2.56% glycation, respectively. The hexane extract of T. ciliata was less toxic to C2C12 cells as compared to the other extracts with CC50 value of 402.16 μg/ml. Only the hexane extract of S. pinnata resulted in glucose utilisation of 28.56% which was higher than that of insulin (26.06%) after 6 hours and is therefore considered as the most potent extract with hypoglycaemic potential in this study. Studies are ongoing aimed at identifying drug candidates in this extract that may be employed in the development of hypoglycaemic, antioxidant, and antiglycation agents