Antioxidative effects of germinated brown rice-derived extracts on H2O2-induced oxidative stress in HepG2 cells

The antioxidant properties of germinated brown rice (GBR) are likely mediated by multiple bioactives. To test this hypothesis, HepG2 cells pretreated with GBR extracts, rich in acylated steryl glycoside (ASG), gamma amino butyric acid GABA), phenolics or oryzanol, were incubated with hydrogen peroxide (H2O2) and their hydroxyl radical (OH•) scavenging capacities and thiobarbituric acid-reactive substances (TBARS) generation were evaluated. Results showed that GBR-extracts increased OH• scavenging activities in both cell-free medium and posttreatment culture media, suggesting that the extracts were both direct- and indirect-acting against OH•. The levels of TBARS in the culture medium after treatment were also reduced by all the extracts. In addition, H2O2 produced transcriptional changes in p53, JNK, p38 MAPK, AKT, BAX, and CDK4 that were inclined towards apoptosis, while GBR-extracts showed some transcriptional changes (upregulation of BAX and p53) that suggested an inclination for apoptosis although other changes (upregulation of antioxidant genes, AKT, JNK, and p38 MAPK) suggested that GBR-extracts favored survival of the HepG2 cells. Our findings show that GBR bioactive-rich extracts reduce oxidative stress through improvement in antioxidant capacity, partly mediated through transcriptional regulation of antioxidant and prosurvival genes

INHIBITION OF STREPTOCOCCUS MUTANS ADHERENCE AND BIOFILM FORMATION ACTIVITIES FROM MELASTOMA MALABATHRICUM SUBFRACTION

Objectives: The objectives of the study were to determine antibacterial, anti-adherence, and antibiofilm activities of Melastoma malabathricum stem bark acetone extract (MMSBAE) subfraction against Streptococcus mutans.Methods: Fraction 9 (F9) from MMSBAE was subfractionated by thin-layer chromatography (TLC) and analyzed for antibacterial activity against S. mutans by TLC-bioautography. Subfraction 12 (SF12) was isolated from F9 followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values.Results: MIC and MBC values were 10 mg/mL and 160 mg/mL, indicating bacteriostatic property of SF12. Time-kill assay analysis confirmed bacteriostatic property of SF12 against S. mutans. Crystal violet staining and glass surface assays were used to determine anti-adherence and antibiofilm activities. Concentrations produced 50% reduction in anti-adherence and antibiofilm activities were 40 mg/mL and 20 mg/mL, respectively. Scanning electron microscopy was performed to visualize the effect of SF12 on S. mutans biofilm structure. SF12 was found to lyse biofilm formation on treated bacteria indicating powerful anticariogenic potential against S. mutans. Analysis by quantitative real-time polymerase chain reaction revealed SF12 at MIC value downregulated biofilm formation genes such as gbpA, brpA, gtfC, and comDE.Conclusion: SF12 showed bacteriostatic activities against S. mutans by inhibiting adherence and biofilm activities

Antidiabetic and Hypolipidemic Activities of Curculigo latifolia Fruit:Root Extract in High Fat Fed Diet and Low Dose STZ Induced Diabetic Rats

Curculigo latifolia fruit is used as alternative sweetener while root is used as alternative treatment for diuretic and urinary problems. The antidiabetic and hypolipidemic activities of C. latifolia fruit:root aqueous extract in high fat diet (HFD) and 40 mg streptozotocin (STZ) induced diabetic rats through expression of genes involved in glucose and lipid metabolisms were investigated. Diabetic rats were treated with C. latifolia fruit:root extract for 4 weeks. Plasma glucose, insulin, adiponectin, lipid profiles, alanine aminotransferase (ALT), gamma glutamyltransferase (GGT), urea, and creatinine levels were measured before and after treatments. Regulations of selected genes involved in glucose and lipid metabolisms were determined. Results showed the significant ( < 0.05) increase in body weight, high density lipoprotein (HDL), insulin, and adiponectin levels and decreased glucose, total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL), urea, creatinine, ALT, and GGT levels in diabetic rats after 4 weeks treatment. Furthermore, C. latifolia fruit:root extract significantly increased the expression of IRS-1, IGF-1, GLUT4, PPAR , PPAR , AdipoR1, AdipoR2, leptin, LPL, and lipase genes in adipose and muscle tissues in diabetic rats. These results suggest that C. latifolia fruit:root extract exerts antidiabetic and hypolipidemic effects through altering regulation genes in glucose and lipid metabolisms in diabetic rats

Thymoquinone prevents β-amyloid neurotoxicity in primary cultured cerebellar granule neurons

Thymoquinone (TQ), a bioactive constituent of Nigella sativa Linn (N. sativa) has demonstrated several neuropharmacological attributes. In the present study, the neuroprotective properties of TQ were investigated by studying its anti-apoptotic potential to diminish β-amyloid peptide 1–40 sequence (Aβ1–40)-induced neuronal cell death in primary cultured cerebellar granule neurons (CGNs). The effects of TQ against Aβ1–40-induced neurotoxicity, morphological damages, DNA condensation, the generation of reactive oxygen species, and caspase-3, -8, and -9 activation were investigated. Pretreatment of CGNs with TQ (0.1 and 1 μM) and subsequent exposure to 10 μM Aβ1–40 protected the CGNs against the neurotoxic effects of the latter. In addition, the CGNs were better preserved with intact cell bodies, extensive neurite networks, a loss of condensed chromatin and less free radical generation than those exposed to Aβ1–40 alone. TQ pretreatment inhibited Aβ1–40-induced apoptosis of CGNs via both extrinsic and intrinsic caspase pathways. Thus, the findings of this study suggest that TQ may prevent neurotoxicity and Aβ1–40-induced apoptosis. TQ is, therefore, worth studying further for its potential to reduce the risks of developing Alzheimer’s disease

A randomised controlled trial on hypolipidemic effects of Nigella Sativa seeds powder in menopausal women

Background: The risk of cardiovascular diseases (CVD) is increased tremendously among menopausal women, and there is an increasing demand for alternative therapies for managing factors like dyslipidemia that contribute to CVD development. Methods: In this study, Nigella sativa was evaluated for its hypolipidemic effects among menopausal women. In a randomised trial, hyperlipidemic menopausal women were assigned to treatment (n = 19) or placebo groups (n = 18), and given N. sativa or placebo for two months after their informed consents were sought. At baseline, blood samples were taken and at one month intervals thereafter until one month after the end of the study. Results: The results showed that N. sativa significantly improved lipid profiles of menopausal women (decreased total cholesterol, low density lipoprotein cholesterol and triglyceride, and increased high density lipoprotein cholesterol) more than the placebo treatment over 2 months of intervention. One month after cessation of treatment, the lipid profiles in the N. sativa-treated group tended to change towards the pretreatment levels. Conclusions: N. sativa is thought to have multiple mechanisms of action and is cost-effective. Therefore, it could be used by menopausal women to remedy hypercholesterolemia, with likely more benefits than with single pharmacological agents that may cause side effects. The use of N. sativa as an alternative therapy for hypercholesterolemia could have profound impact on the management of CVD among menopausal women especially in countries where it is readily available

Effect of kenaf seed oil from different ways of extraction towards ovarian cancer cells.

Kenaf (Hibiscus cannabinus) from the family of Malvaceae is a valuable fibre plant native to India and Africa. Kenaf is composed of various active components including tannins, saponins, polyphenolics, alkaloids, essential oils and steroids. It has been used to treat bruises, bilious conditions, fever and puerperium. Nevertheless, the anti-cancer properties of kenaf seed oil have not yet been investigated. In this study, kenaf seed oils obtained by Sonication, Soxhlet and supercritical carbon dioxide fluid extraction (SFE) with 9 different combinations of pressure (bars) and temperature (°C) (200/40, 200/60, 200/80, 400/40, 400/60, 400/80, 600/40, 600/60 and 600/80) were investigated for the cytotoxicities. All the oils were cytotoxic towards ovarian cancer (CaOV3) and colon cancer (HT29) cell lines in a dose dependent manner as detected by using the MTT assay and trypan blue dye exclusion method. Oil from Sonication was the most cytotoxic towards CaOV3 cell line. Treated cells exhibited characteristics of apoptosis such as chromatin condensation and nuclear fragmentation. In conclusion, kenaf seed oils from the three extractions were cytotoxic towards CaOV3 cell line in a dose-dependent manner possibly via the induction of apoptosis. In considering the safety of the product, SFE technology is a better alternative extraction method that is suitable in kenaf seed oil extraction

Effects of supercritical fluid-extracted Hibiscus cannabinus L. seed oil on colon cancer in vitro and in vivo

Kenaf (Hibiscus cannabinus) from the family Malvaceae, is a valuable fiber plant native to India and Africa, and is currently planted as the fourth commercial crop in Malaysia. Kenaf seed oil contains alpha-linolenic acid, phytosterol such as β-sitosterol, vitamin E and other antioxidants with chemopreventive properties. The present study evaluated cytotoxicity towards human colorectal cancer cell line (HT29) and cancer chemopreventive properties of kenaf seed oil from supercritical carbon dioxide fluid extraction (KSO-SFE). Kenaf seed oil was extracted via supercritical carbon dioxide fluid (SFE) at 9 different permutations of parameters based on range of pressure (200-600 bars) and temperature (40-80°C): 200/40, 200/60, 200/80, 400/40, 400/60, 400/80,600/40, 600/60 and 600/80. All the nine KSO-SFE were screened for cytotoxicity towards human colorectal cancer cell line (HT29) and mouse embryonic fibroblast cell line (NIH/3T3) using MTS assay. KSO-SFE of 600/40 showed the strongest cytotoxicity towards HT29 with IC50 of 200 μg/ml. Nevertheless, IC50 for NIH/3T3 was not detected even at the highest concentration of KSO-SFE employed. Cell cycle analysis exhibited a significant increase in the number of KSO-SFE-treated cells at sub- G1 phase, indicating the induction of apoptosis by the extract. The induction of apoptosis was further confirmed by Annexin V/PI and AO/PI staining. For the chemopreventive properties of KSO-SFE, 60 male Sprague Dawley rats were randomly assigned to 6 groups. All groups were induced with azoxymethane (AOM) except for the negative control (Group 1). They were 1) negative control group, 2) positive control group (without any treatment), 3) vehicle control group (administered with emulsifier (Tween 80), 4) group treated with 500 mg/kg body weight KSO-SFE; 5) group treated with 1000 mg/kg body weight KSO-SFE and 6) group treated with 1500 mg/kg body weight KSO-SFE. The animals were injected subcutaneously once a week for 2 weeks with 15 mg/kg body weight of AOM at 7 weeks of age. Rats were euthanized after 90 days of the experiment. There was no significant difference in weight gain among the groups. Number of aberrant crypt foci (ACF) ranged from 84.4 ± 4.43 to 179.5 ± 12.78 in Group 2, 3, 4, 5 and 6. ACF were reduced by 45.3%, 51.4% and 53.1% in rats fed with 500, 1000 and 1500 mg/kg body weight of KSO-SFE, respectively, compared to the positive control group (p<0.05). For ACF multiplicity, ACF with 4, 5 or more crypts were significantly lower (p<0.05) in rats fed with KSO-SFE compared to the positive control group. The findings indicate that KSO-SFE reduced AOM–induced ACF in Sprague Dawley male rats. The effects of KSO-SFE on fifteen genes involved in colon carcinogenesis were analyzed on AOM-induced rats using GenomeLabGeXP genetic system. It shows that treatment with KSO-SFE increased the expression of tumor suppressor genes (APC and p53), reduced the expression of tumor marker genes (COX-2and β-catenin) and did not change the expression of large tumor suppressor and TNF receptor genes compared to the positive control group (p<0.05). KSO-SFE has also shown to activate the apoptotic pathway by up regulating the expression of caspases (caspase 9, caspase 2 and caspase 3) and pro-apoptotic genes (bax and bad), and down regulating the expression of anti-apoptotic gene (bcl-2). Treatment with KSO-SFE also affects the cell cycle genes with increased expression of cell cycle inhibitor (p21, cip1) and decreased expression of cyclin D1. Assessment of toxicity of KSO-SFE at 500, 1000 and 1500 mg/kg body weight/day towards Sprague Dawley rats was also performed. The parameters for toxicity include body and organ weight, haematology,clinical chemistry, pathology and expression of toxicity-related genes. No mortality or treatment-related adverse effects were observed at all doses throughout the period of 90 days. All the parameters were in normal range. Low creatinine level at all doses and low total cholesterol level at 1000 and 1500 mg/kg body weight of KSO-SFE were noted but insignificant. Further analysis using GenomeLabGeXP genetic system on the liver tissues showed the expression of genes involved in xenobiotic metabolism, DNA damage, cell cycle arrest and apoptosis was at normal level. In short, The No Observed Adverse Effect Level (NOAEL) for KSO-SFE at 1500 mg/kg body weight/day. In conclusion, data from this study demonstrate the potential of KSO-SFE as a chemopreventive agent against colon cancer

Antidiabetic and hypolipidemic activities of Curculigo latifolia fruit: root extract in high fat fed diet and low dose STZ induced diabetic rats

Curculigo latifolia fruit is used as alternative sweetener while root is used as alternative treatment for diuretic and urinary problems. The antidiabetic and hypolipidemic activities of C. latifolia fruit:root aqueous extract in high fat diet (HFD) and 40 mg streptozotocin (STZ) induced diabetic rats through expression of genes involved in glucose and lipid metabolisms were investigated. Diabetic rats were treated with C. latifolia fruit:root extract for 4 weeks. Plasma glucose, insulin, adiponectin, lipid profiles, alanine aminotransferase (ALT), gamma glutamyltransferase (GGT), urea, and creatinine levels were measured before and after treatments. Regulations of selected genes involved in glucose and lipid metabolisms were determined. Results showed the significant () increase in body weight, high density lipoprotein (HDL), insulin, and adiponectin levels and decreased glucose, total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL), urea, creatinine, ALT, and GGT levels in diabetic rats after 4 weeks treatment. Furthermore, C. latifolia fruit:root extract significantly increased the expression of IRS-1, IGF-1, GLUT4, PPARα, PPARγ, AdipoR1, AdipoR2, leptin, LPL, and lipase genes in adipose and muscle tissues in diabetic rats. These results suggest that C. latifolia fruit:root extract exerts antidiabetic and hypolipidemic effects through altering regulation genes in glucose and lipid metabolisms in diabetic rats

Therapeutic Potential of Honey and Propolis on Ocular Disease

Honey and propolis have recently become the key target of attention for treating certain diseases and promoting overall health and well-being. A high content of flavonoids and phenolic acids found in both honey and propolis contributes to the antioxidant properties to scavenge free radicals. Honey and propolis also exhibited antibacterial effects where they act in two ways, namely the production of hydrogen peroxide (H2O2) and gluconic acids following the enzymatic activities of glucose oxidase, which exerts oxidative damage on the bacteria. Additionally, the anti-inflammatory effects of honey and propolis are mainly by reducing proinflammatory factors such as interleukins and tumor necrosis factor alpha (TNF-&alpha;). Their effects on pain were discovered through modulation at a peripheral nociceptive neuron or binding to an opioid receptor in the higher center. The aforementioned properties of honey have been reported to possess potential therapeutic topical application on the exterior parts of the eyes, particularly in treating conjunctivitis, keratitis, blepharitis, and corneal injury. In contrast, most of the medicinal values of propolis are beneficial in the internal ocular area, such as the retina, optic nerve, and uvea. This review aims to update the current discoveries of honey and propolis in treating various ocular diseases, including their antioxidant, anti-inflammatory, antibacterial, and anti-nociceptive properties. In conclusion, research has shown that propolis and honey have considerable therapeutic promise for treating various eye illnesses, although the present study designs are primarily animal and in vitro studies. Therefore, there is an urgent need to translate this finding into a clinical setting

Silver nanoparticles Clinacanthus nutans leaves extract induced apoptosis towards oral squamous cell carcinoma cell lines

Purpose:The purpose of this study was to investigate apoptotic activity of silver nanoparticle Clinacanthus nutans(AgNps-CN) towards HSC-4 cell lines (oral squamous cell carcinoma cell lines).Methods:Methods involved were MTT assay (cytotoxic activity), morphological cells analysis, flow cytometry and cell cycle analysis and western blot.Results:MTT assay revealed IC50concentration was 1.61mg/mL, 3T3-L1 cell lines were used to deter-mine whether AgNps-CN is cytotoxic to normal cells. At the highest concentration (3mg/mL), no cyto-toxic activity has been observed. Flow cytometry assay revealed AgNps-CN caused apoptosis effects towards HSC-4 cell lines with significant changes were observed at G1 phase when compared with untreated cells. Morphological cells analysis revealed that most of the cells exhibit apoptosis characteristics rather than necrosis. Protein study revealed that ratio of Bax/Bcl-2 increased mainly due to down-regulation of Bcl-2 expression.Conclusion:AgNps-CN have shown potential in inhibiting HSC-4 cell lines. IC50was low compared to few studies involving biosynthesized of silver nanoparticles. Apoptosis effects were shown towardsHSC-4 cell lines by the increased in Bax/Bcl-2 protein ratio. Further study such as PCR or in vivo studies are required