Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots

In this study, the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated. From MTT assays, the concentration of the shoot extracts that maintained 50% cell viability (IC50) was 1.7 mg/ml. Cell viability was kept above 90% at both 0.4 mg/ml and 0.6 mg/ml of the extracts. The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays. The microarray data were validated using real-time qRT–PCR. A total of 246, 696 and 4503 genes were significantly regulated (P < 0.01) by at least 1.5-fold in response to 0.4, 0.6 and 1.7 mg/ml of the extracts, respectively. Mutually regulated genes in response to the three concentrations included CDKN3, LOC100289612, DHFR, VRK1, CDC6, AURKB and GABRE. Genes like CYP24A1, BRCA1, AURKA, CDC2, CDK2, CDK4 and INSR were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml. However, the expression of genes including LGR5, IGFBP3, RB1, IDE, LDLR, MTTP, APOB, MTIX, SOD2 and SOD3 were exclusively regulated at the IC50 concentration. In conclusion, low concentrations of the extracts were able to significantly regulate a sizable number of genes. The type of genes that were expressed was highly dependent on the concentration of the extracts used

Gene expression profiles in human HepG2 cells treated with extracts of the Tamarindus indica fruit pulp

Tamarindus indicaL. (T. indica) or locally known as asam jawa belongs to the family of Leguminosae. The fruit pulp had been reported to have antioxidant activities and possess hypolipidaemic effects. In this study, we attempted to investigate the gene expression patterns in human hepatoma HepG2 cell line in response to treatment with low concentration of the fruit pulp extracts. Microarray analysis using Affymetrix Human Genome 1.0 S.T arrays was used in the study. Microarray data were validated using semi-quantitative RT–PCR and real-time RT–PCR. Amongst the significantly up-regulated genes were those that code for the metallothioneins (MT1M, MT1F, MT1X) and glutathione S-transferases (GSTA1, GSTA2, GST02) that are involved in stress response. APOA4, APOA5, ABCG5 and MTTP genes were also significantly regulated that could be linked to hypolipidaemic activities of the T. indica fruit pulp

Evaluation of bioactivities of tamarindus indica and the effects of the antioxidant-rich extracts on the transcriptome profiles in liver hepg2 cells / Nurhanani Razali

Tamarindus indica L. (T. indica) or locally known as “asam jawa”, belongs to the family Leguminosae and grows naturally in many tropical and sub-tropical regions. In this study, the antioxidant activities of T. indica leaf, seed, vein and skin extracted with methanol, ethyl acetate and hexane were investigated. In addition, the effects of the antioxidant-rich crude and fractionated extracts of T. indica on lipid peroxidation, antioxidant enzyme activities and gene expression patterns were investigated in liver HepG2 cells. Amongst the various parts of T. indica, the methanol leaf and seed extracts showed high phenolic content and antioxidant activities. The crude methanol extracts of T. indica leaf and seed were subjected to bioactivity-guided fractionation. Fraction 10 (F10) and fraction 3 (F3), obtained from the methanol leaf and seed extracts, respectively, showed the highest phenolic content and antioxidant activities. When treated on HepG2 cells, F10 and F3 exhibited potent cytotoxic effect and antioxidant activities and were able to inhibit lipid peroxidation and enhanced superoxide dismutase, catalase and glutathione peroxidase activities. Overall, fractionation of the methanol leaf and seed extracts gave higher phenolic content, antioxidative protection and cytotoxicity compared to the respective crude extracts. Amongst all extracts, F10 showed the most potent antioxidant activities. Analyses of polyphenols in the methanol leaf extract using UHPLC revealed the presence of gallic acid, epicatechin, caffeic acid, p-coumaric acid, ferulic acid, ellagic acid, quercetin, apigenin, isorhamnetin and an unknown compound. UHPLC analyses of the methanol seed extract revealed the presence of catechin, procyanidin B2, caffeic acid, ferulic acid, chloramphenicol, myricetin, morin, quercetin, apigenin and iv kaempferol. NMR analyses characterised F3 as caffeic acid, while a novel compound was identified in F10. cDNA microarray analysis revealed that 207 genes were significantly regulated by at least 1.5-fold (p < 0.05) in leaf-treated HepG2 cells. When the significantly regulated genes were analysed using Ingenuity Pathway Analysis (IPA) software, “Lipid Metabolism, Small Molecule Biochemistry, Hematological Disease’ was the top network affected by the leaf extract, with a score of 36. The top predicted canonical pathway affected by the leaf extract was the coagulation system (P < 2.80 x 10-6) involving five genes namely KNG1, SERPINC1, SERPIND1, SERPINE1, FGG, FGA. In seed-treated HepG2 cells, 167 genes were significantly up-regulated while 80 genes were down-regulated in response to the IC20 treatment. IPA analyses showed that “Free Radical Scavenging, Hematological System Development and Function, Inflammatory Response” was the top network affected by the seed extract, with a score of 45. The top predicted canonical pathway affected by the seed extract was the NRF2-mediated oxidative stress response (P < 2.84 x 10-9) involving 10 genes namely AKT1, DNAJC10, GPX2, GSTA4, MAPK8, MGST2, PTPLAD1, SOD1, TXNRD1, USP14. Similar pattern of expression was observed for selected proteins. The promising therapeutic potency of T. indica particularly the leaves and the seeds reported in this study implies the potential of this plant as an alternative source of natural antioxidant agent. This study also provides further scientific evidence to support the medicinal uses of this plant

Correlation between body composition and cardiorespiratory fitness in female adults of different Body Mass Index

BACKGROUND: Obesity has become a major public concern as it leads to significant health risk and economic burden especially to the developing countries. The prevalence of obesity is also significantly higher among the female adults. It is also known that low fitness level is a powerful and independent risk factor for premature mortality. However, the understanding of the relationship between overall body composition indices and cardiorespiratory fitness in adults remains limited. This study aimed to investigate the correlation between body composition with cardiorespiratory fitness among normal weight, overweight and obese in female adults. METHODS: Eighty-one female participants aged between 18 to 59 years who cleared the inclusion and exclusion criteria were divided into three groups; normal weight (N.=23), overweight (N.=29) and obese (N.=29). They went through bio-electrical impedance analysis measurement and cardiopulmonary fitness testing using ramp protocol in Sport Medicine human performance lab. RESULTS: It was noted that there was an inverse strong strength significant correlation between VO 2 max and Body Mass Index (r=-0.588, P<0.001, Waist Hip Ratio (r=-0.596, P<0.001), Percentage Body Fat (r=-0.684, P<0.001) and Body Fat Mass (r=-0.667, P<0.001). However, skeletal muscle mass showed no significant correlation with VO 2 max. There was also a high significant difference among the three groups in term of VO 2 max (F=7.683, P=0.001) after alleviating the age factor. CONCLUSIONS: In conclusion, our study showed significant correlation between VO 2 and body composition particularly percentage of body fat (%) and body fat mass (kg). It will be more convenience and cost-effective especially in developing countries, if body composition can be used as an independent entity to predict cardiorespiratory fitness among individual. Furthermore, cardiorespiratory exercise testing showed significant differences of VO 2 max among 3 groups. This will be beneficial if the VO 2 max is used as a tool to predict cardiovascular risk. Therefore, we would suggest in-depth studies to be done in the future to provide more knowledge regarding this important topic on a bigger scale subjects. © 2018 EDIZIONI MINERVA MEDICA

Antioxidant-rich leaf extract of Barringtonia racemosa significantly alters the in vitro expression of genes encoding enzymes that are involved in methylglyoxal degradation III

Background Barringtonia racemosa is a medicinal plant belonging to the Lecythidaceae family. The water extract of B. racemosa leaf (BLE) has been shown to be rich in polyphenols. Despite the diverse medicinal properties of B. racemosa, information on its major biological effects and the underlying molecular mechanisms are still lacking. Methods In this study, the effect of the antioxidant-rich BLE on gene expression in HepG2 cells was investigated using microarray analysis in order to shed more light on the molecular mechanism associated with the medicinal properties of the plant. Results Microarray analysis showed that a total of 138 genes were significantly altered in response to BLE treatment (p < 0.05) with a fold change difference of at least 1.5. SERPINE1 was the most significantly up-regulated gene at 2.8-fold while HAMP was the most significantly down-regulated gene at 6.5-fold. Ingenuity Pathways Analysis (IPA) revealed that “Cancer, cell death and survival, cellular movement” was the top network affected by the BLE with a score of 44. The top five canonical pathways associated with BLE were Methylglyoxal Degradation III followed by VDR/RXR activation, TR/RXR activation, PXR/RXR activation and gluconeogenesis. The expression of genes that encode for enzymes involved in methylglyoxal degradation (ADH4, AKR1B10 and AKR1C2) and glycolytic process (ENO3, ALDOC and SLC2A1) was significantly regulated. Owing to the Warburg effect, aerobic glycolysis in cancer cells may increase the level of methylglyoxal, a cytotoxic compound. Conclusions BLE has the potential to be developed into a novel chemopreventive agent provided that the cytotoxic effects related to methylglyoxal accumulation are minimized in normal cells that rely on aerobic glycolysis for energy supply

Analyses of antioxidant status and nucleotide alterations in genes encoding antioxidant enzymes in patients with benign and malignant thyroid disorders

Background Synthesis of thyroid hormones and regulation of their metabolism involve free radicals that may affect redox balance in the body. Thyroid disorders causing variations in the levels of thyroid hormones may alter cellular oxidative stress. The aim of this study was to measure the antioxidant activities and biomarkers of oxidative stress in serum and red blood cells (RBC) of patients with benign and malignant thyroid disorders and to investigate if changes in the antioxidant activities in these patients were linked to alterations in genes encoding the antioxidant enzymes. Methods Forty-one patients with thyroid disorders from University of Malaya Medical Centre were recruited. They were categorised into four groups: multinodular goitre (MNG) (n = 18), follicular thyroid adenoma (FTA) (n = 7), papillary thyroid cancer (PTC) (n = 10), and follicular thyroid cancer (FTC) (n = 6). Serum and RBC of patients were analysed for antioxidant activities, antioxidant enzymes, and biomarkers of oxidative stress. Alterations in genes encoding the antioxidant enzymes were analysed using whole exome sequencing and PCR–DNA sequencing. Results Patients with thyroid disorders had significantly higher serum superoxide dismutase (SOD) and catalase (CAT) activities compared to control, but had lower activities in RBC. There were no significant changes in serum glutathione peroxidase (GPx) activity. Meanwhile, GPx activity in RBC was reduced in PTC and FTC, compared to control and the respective benign groups. Antioxidant activities in serum were decreased in the thyroid disorder groups when compared to the control group. The levels of malondialdehyde (MDA) were elevated in the serum of FTA group when compared to controls, while in the RBC, only the MNG and PTC groups showed higher MDA equivalents than control. Serum reactive oxygen species (ROS) levels in PTC group of both serum and RBC were significantly higher than control group. Whole exome sequencing has resulted in identification of 49 single nucleotide polymorphisms (SNPs) in MNG and PTC patients and their genotypic and allelic frequencies were calculated. Analyses of the relationship between serum enzyme activities and the total SNPs identified in both groups revealed no correlation. Discussion Different forms of thyroid disorders influence the levels of antioxidant status in the serum and RBC of these patients, implying varying capability of preventing oxidative stress. A more comprehensive study with a larger target population should be done in order to further evaluate the relationships between antioxidant enzymes gene polymorphisms and thyroid disorders, as well as strengthening the minor evidences provided in literatures

Identification of Proteins of Altered Abundance in Oil Palm Infected with Ganoderma boninense

Basal stem rot is a common disease that affects oil palm, causing loss of yield and finally killing the trees. The disease, caused by fungus Ganoderma boninense, devastates thousands of hectares of oil palm plantings in Southeast Asia every year. In the present study, root proteins of healthy oil palm seedlings, and those infected with G. boninense, were analyzed by 2-dimensional gel electrophoresis (2-DE). When the 2-DE profiles were analyzed for proteins, which exhibit consistent significant change of abundance upon infection with G. boninense, 21 passed our screening criteria. Subsequent analyses by mass spectrometry and database search identified caffeoyl-CoA O-methyltransferase, caffeic acid O-methyltransferase, enolase, fructokinase, cysteine synthase, malate dehydrogenase, and ATP synthase as among proteins of which abundances were markedly altered

LC-MS/MS Proteomic Study of MCF-7 Cell Treated with Dox and Dox-Loaded Calcium Carbonate Nanoparticles Revealed Changes in Proteins Related to Glycolysis, Actin Signalling, and Energy Metabolism

One of the most prevalent death causes among women worldwide is breast cancer. This study aimed to characterise and differentiate the proteomics profiles of breast cancer cell lines treated with Doxorubicin (DOX) and Doxorubicin-CaCO3-nanoparticles (DOX-Ar-CC-NPs). This study determines the therapeutic potential of doxorubicin-loaded aragonite CaCO3 nanoparticles using a Liquid Chromatography/Mass Spectrometry analysis. In total, 334 proteins were expressed in DOX-Ar-CC-NPs treated cells, while DOX treatment expressed only 54 proteins. Out of the 334 proteins expressed in DOX-CC-NPs treated cells, only 36 proteins showed changes in abundance, while in DOX treated cells, only 7 out of 54 proteins were differentially expressed. Most of the 30 identified proteins that are differentially expressed in DOX-CC-NPs treated cells are key enzymes that have an important role in the metabolism of carbohydrates as well as energy, including: pyruvate kinase, ATP synthase, enolase, glyceraldehyde-3-phosphate dehydrogenase, mitochondrial ADP/ATP carrier, and trypsin. Other identified proteins are structural proteins which included; Keratin, α- and β-tubulin, actin, and actinin. Additionally, one of the heat shock proteins was identified, which is Hsp90; other proteins include Annexins and Human epididymis protein 4. While the proteins identified in DOX-treated cells were tubulin alpha-1B chain and a beta chain, actin cytoplasmic 1, annexin A2, IF rod domain-containing protein, and 78 kDa glucose-regulated protein. Bioinformatics analysis revealed the predicted canonical pathways linking the signalling of the actin cytoskeleton, ILK, VEGF, BAG2, integrin and paxillin, as well as glycolysis. This research indicates that proteomic analysis is an effective technique for proteins expression associated with chemotherapy drugs on cancer tumours; this method provides the opportunity to identify treatment targets for MCF-7 cancer cells, and a liquid chromatography-mass spectrometry (LC-MS/MS) system allowed the detection of a larger number of proteins than 2-DE gel analysis, as well as proteins with maximum pIs and high molecular weight

Molecular investigation of carrageenan production in Kappaphycus alvarezii in different culture conditions: a proteomic approach

Carrageenan is a polysaccharide extracted from different species of red seaweed for use in pharmaceuticals and cosmetics industries. In Malaysia, κ-carrageenan is obtained through extraction from the tropical red seaweed Kappaphycus alvarezii. The use of tissue culture techniques in the propagation of K. alvarezii has proven to be effective in solving cultivation problems and produced high-quality seedlings and providing a sustainable source of better quality carrageenan. In order to understand the molecular mechanisms behind that, a proteomic investigation was conducted comparing the changes in protein expression in tissue-cultured and liquid-cultured K. alvarezii after 60 days of cultivation. Proteomic analysis was used to study the changes in the protein expression level between liquid-cultured and tissue-cultured of the red seaweed K. alvarezii. A total of 45 protein spots were found to be significantly different in their densities and three proteins, namely β-amylase, NAD-dependent sugar epimerase and B-phycoerythrin, showed a consistent pattern of upregulation in ELISA analyses, hence validating the 2-DE profiles. Changes in the proteins expression level were noticed in proteins related to energy production, metabolism and cellular maintenance. The protein changes in tissue-cultured seaweed possibly play an essential role in the production of carrageenan