Rukkumani Rajagopalan

Antiproliferative and Apoptotic Effects of Sesbania grandiflora Leaves in Human Cancer Cells

Natural phytochemicals and their derivatives are good drug candidates for anticancer therapeutic approaches against multiple targets. We report here the initial findings from our studies on the anticancer properties of the leaves of the medicinal plant Sesbania grandiflora. In the current study, five different solvent fractions from the leaves of S. grandiflora were tested on cancer cell lines such as MCF-7, HepG2, Hep-2, HCT-15, and A549. The methanolic fraction of S. grandiflora was found to exert potent antiproliferative effects especially in the human lung cancer cell line, A549. Caspase 3 was activated in the methanolic fraction treated A549 cells thereby leading to cell death by apoptosis. DAPI staining, DNA laddering, and decrease in mitochondrial membrane potential further confirmed the apoptotic mode of cell death. The high levels of ROS intermediates as evidenced by DCF-DA staining could have played a role in the apoptotic induction. Decrease in levels of cyclin D1 and decrease in the activation of NFkB were observed in A549 cells on treatment with methanolic fraction, giving a hint on the possible mechanism of action. These results prove that the medicinal plant S. grandiflora can be explored further for promising candidate molecules to combat cancer, especially lung cancer.

Hepatoprotective role of bis-demethoxy curcumin analog on the expression of matrix metalloproteinase induced by alcohol and polyunsaturated fatty acid in rats

Liver fibrosis is one of the major health problems worldwide. Chronic alcohol abuse is one of the main causes of fibrosis. Ingestion of polyunsaturated fatty acids (PUFA) along with alcohol further aggravates the toxicity of alcohol. Fibrosis results due to increased deposition of extra cellular matrix (ECM). The degree of abnormal ECM degradation depends on the ratio of active matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The present work studied the influence of bis-desmethoxy curcumin analog (BDMC-A) on the expression of MMPs and TIMPs during alcohol and ΔPUFA induced liver toxicity. Male albino Wistar rats were used for the study. The MMP expression was found to be increased in alcohol as well as ΔPUFA treated rats and decreased in alcohol + ΔPUFA treated rats. The levels of TIMPs and the collagen were increased in alcohol, ΔPUFA, and alcohol + ΔPUFA groups. Administration of BDMC-A significantly decreased the levels of collagen and TIMPs; and positively modulated the expression of MMPs. From this study, it is concluded that BDMC-A influences MMPs, TIMPs expression, and acts as an efficient anti-fibrotic agent.

Influence of stabilizers on the production of disulfiram-loaded poly(lactic-co-glycolic acid) nanoparticles and their anticancer potential

AIM Our hypothesis was to prove that surface modifiers themselves can be used as stabilizers and that their entrapment efficiency is directly influenced by the type of stabilizers used. MATERIALS & METHODS Particle size and the polydispersity index of the nanoparticles (NPs) were measured by dynamic light scattering, whereas the morphology of the NPs was studied by scanning electron microscopy. Percentage nanoparticle yield, entrapment efficiency and drug loading capacity were measured by ultraviolet absorbance. The physical rigidity, robustness and drug releasing capability of these NPs were also assessed. CONCLUSION Physiochemical characterization and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay suggest that polysorbate 80 has the dual capability of being a stabilizer and a surface modifier in addition to having better drug entrapment properties than Pluronic® 188. Disulfiram, the drug that was loaded on these NPs, is also observed for the first time to show significant anticancer potential against hepatocellular carcinoma (Hep3B) cell lines.

Hepatoprotective role of kaempferol during alcohol- and ΔPUFA-induced oxidative stress.

Abstract Background: The present study aimed to analyze the effect of kaempferol on oxidative stress induced by alcohol and thermally oxidized polyunsaturated fatty acid (ΔPUFA) in male albino Wistar rats. Methods: The rats were divided into four groups. The animals in group 1 served as the normal group (standard diet), group 2 served as the hepatotoxic group (alcohol+ΔPUFA), group 3 served as the treated group (alcohol+ΔPUFA+kaempferol), and group 4 served as kaempferol control. The levels of marker enzyme γ-glutamyl transferase (GGT), lipid peroxidation markers [thiobarbituric acid reactive substances (TBARS) and lipid hydroperoxides (LH)], enzymatic antioxidants (catalase, superoxide dismutase, and glutathione peroxidase), and nonenzymatic antioxidants (reduced glutathione, vitamin E, and vitamin C) were analyzed in liver to evaluate the effects of kaempferol. Results: The levels of GGT, TBARS, and LH were significantly increased in liver of the alcohol+ΔPUFA group and were found to be reduced on treatment with kaempferol. The levels of both enzymatic and nonenzymatic antioxidants were decreased in liver of the alcohol+ΔPUFA group and were found to be restored on treatment with kaempferol. Conclusions: From the results obtained, we conclude that kaempferol protects the liver against alcohol- and ΔPUFA- induced oxidative stress.

Protective role of ferulic acid on carbon tetrachloride-induced hyperlipidemia and histological alterations in experimental rats.

Abstract Background: The present study aimed to evaluate the antihyperlipidemic effect of ferulic acid (FA) on carbon tetrachloride (CCl4)-treated rats. Methods: Female albino rats of the Wistar strain were used in the present study. The rats were divided into four groups: groups 1 and 4 received physiological saline (3 mL/kg body weight/week) by subcutaneous injection, whereas groups 2 and 3 received a subcutaneous injection of CCl4 (3 mL/kg body weight/week) for a total period of 12 weeks. In addition, groups 3 and 4 were administered FA (20 mL/kg body weight) every day for the last 90 days. Results: The results showed significantly (p≤0.05) elevated levels of cholesterol, triglycerides (TG), and free fatty acids (FFA) in the liver and kidney of CCl4-treated rats as compared with those of the controls. In addition, the levels of cholesterol, FFA, phospholipids (PL), and TG were elevated significantly in the circulation. Administration of FA effectively reduced these levels of lipids in the plasma, liver, and kidney of CCl4-treated rats. The PL level was significantly decreased in the liver and kidney of CCl4-treated rats and was positively modulated by FA treatment. Our histopathological observations were also in correlation with the biochemical parameters. Conclusions: From the results obtained, we could conclude that FA effectively protects the system against hyperlipidemia and may be an effective therapeutic agent for the treatment of this disorder.

Phytochemical screening and analysis of antioxidant properties of aqueous extract of wheatgrass.

OBJECTIVE To screen the phytochemical constituents and study antioxidant properties of the aqueous extract of the wheatgrass. METHODS The current study was focused on broad parameters namely, phytochemical analysis, gas chromatography-mass spectrometry analysis and antioxidant properties in order to characterize the aqueous extract of wheatgrass as a potential free radical quencher. RESULTS The phytochemical screening of the aqueous extract of wheatgrass showed the presence of various secondary metabolites but the absence of sterols and quinone in general. Wheatgrass was proved to be an effective radical scavenger in all antioxidant assays. The gas chromatography-mass spectrometry analysis confirmed the presence of diverse category of bioactive compounds such as squalene, caryophyllene and amyrins in varying percentage. CONCLUSIONS From the results obtained, we conclude that wheatgrass aqueous extract contains various effective compounds. It is a potential source of natural antioxidants. Further analysis of this herb will help in finding new effective compounds which can be of potent use in pharmacological field.

Methanol extract of wheatgrass induces G1 cell cycle arrest in a p53-dependent manner and down regulates the expression of cyclin D1 in human laryngeal cancer cells-an in vitro and in silico approach

Background: Deregs been implicated in the malignancy of cancer. Since many years investigation on the traditional herbs has been the focus to develop novel and effective drug for cancer remedies. Wheatgrass is a medicinal plant, used in folk medicine to cure various diseases. The present study was undertaken to gain insights into antiproliferative effect of methanol extract of wheatgrass. Materials Methods: Cell viability was assessed via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Lactate Dehydrogenase assays. Cell cycle was analyzed by flow cytometry. Western blot was performed to determine the p53 and cyclin D1 levels. In silico docking interaction of the 14 active components (identified by high-performance liquid chromatography/gas chromatography-mass spectroscopy) of the methanol extract was tested with cyclin D1 (Protein Data Bank ID: 2W96) and compared with the reference cyclin D1/Cdk4 inhibitor. Results: Methanol extract of wheatgrass effectively reduced the cell viability. The cell cycle analysis showed that the extract treatment caused G1 arrest. The level of cyclin D1 was decreased, whereas p53 level was increased. Molecular docking studies revealed interaction of seven active compounds of the extract with the vital residues (Lys112/Glu141) of cyclin D1. Conclusion: These findings indicate that the methanol extract of wheatgrass inhibits human laryngeal cancer cell proliferation via cell cycle G1 arrest and p53 induction. The seven active compounds of the extract were also found to be directly involved in the inhibition of cyclin D1/Cdk4 binding, thus inhibiting the cell proliferation.

Inhibition of metastasis and angiogenesis in Hep-2 cells by wheatgrass extract – an in vitro and in silico approach

Abstract Metastasis is the major hindrance in the treatment of all cancers, including laryngeal squamous cell carcinoma. Intensive researches are under way to identify the effective natural polyphenols with anti-metastatic ability for cancer treatment. Wheatgrass, an herbal plant has been reported to show anticancer effects. Hence, in this study, we aimed to analyze the anti-metastatic effect of methanol extract of wheatgrass (MEWG). The levels of metastatic marker proteins were determined by western blot. PI3K and AKT levels were determined by real time (RT)-PCR analysis. In silico molecular docking was done to check the interaction of the 14 components (identified by HPLC/GCMS) of MEWG with PI3K and AKT. MEWG effectively decreased the metastatic protein expressions, namely VEGF, MMP-9 and COX-2 and increased TIMP-2. RT-PCR results showed reduced m-RNA levels of both PI3K and AKT when compared to control. Molecular docking studies revealed interaction of most of the identified compounds of the extract with the important residues of PI3K and AKT. These findings indicate that MEWG inhibits metastasis and angiogenesis in Hep-2 cells possibly via PI3K/AKT due to the cumulative effect of polyphenols and other constituent present in extract. The compounds of the extract were also found to be directly involved in inhibition of AKT/PI3K, thus could help to restrain metastasis.

Hypoglycaemic role of wheatgrass and its effect on carbohydrate metabolic enzymes in type II diabetic rats.

Diabetes mellitus (DM) is a leading cause of morbidity and mortality in the world. Insulin resistance and insulin insufficiency is the major factor for the prognosis of type II diabetes. Consistent high glucose level leads to multiple secondary complications in diabetic patients. Hence, hypoglycaemic drugs are of significance for reducing the risk of secondary complications in type II diabetes. Various hypoglycaemic drugs are already available in the market, but they are associated with several side effects. Therefore, traditional herbs have emerged as safer alternative for effective hypoglycaemic treatment. The juvenile grass of common wheat is known as wheatgrass (WG). It is commonly used as a health drink and has potent antioxidant efficacy. It has been used to cure DM in folk medicine. The current study was planned to test the hypoglycaemic effect and pathways regulated by WG on DM. We analysed the glucose and insulin levels in plasma, the activity of glucose oxidative enzymes, hexokinase and glucose 6 phosphate dehydrogenase, in serum and glycogen levels in liver of the male albino Wistar rats. Activity of glucose oxidative enzymes and the levels of insulin and liver glycogen were decreased in rats with diabetes, but they were reversed on treatment with WG. Hence, we conclude that WG can act as a potent anti-hyperglycaemic agent.

Stabilizers influence drug–polymer interactions and physicochemical properties of disulfiram-loaded poly-lactide-co-glycolide nanoparticles

Aim: Stabilizers are known to be an integral component of polymeric nanostructures. Ideally, they manipulate physicochemical properties of nanoparticles. Based on this hypothesis, we demonstrated that disulfiram (drug) and Poly-lactide-co-glycolide (polymer) interactions and physicochemical properties of their nanoparticles formulations are significantly influenced by the choice of stabilizers. Methodology: Electron microscopy, differential scanning calorimetry, x-ray diffraction, Raman spectrum analysis, isothermal titration calorimetry and in silico docking studies were performed. Results & discussion: Polysorbate 80 imparted highest crystallinity while Triton-X 100 imparted highest rigidity, possibly influencing drug bioavailability, blood-retention time, cellular uptake and sustained drug release. All the molecular interactions were hydrophobic in nature and entropy driven. Therefore, polymeric nanoparticles may be critically manipulated to streamline the passive targeting of drug-loaded nanoparticles.