ABSTRACT
Inhibition of the 26S proteasome is an attractive approach for anticancer therapy. Proteasome inhibitors are known to selectively target cancer cells and make them more sensitive to chemotherapeutic agents. Murraya koenigii is a medicinally important herb of Asian origin and a rich source of bioactive compounds such as flavonoids and alkaloids. In the present study, we investigated the proteasome inhibitory and apoptotic effect of M. koenigii leaf extract in vivo in a xenograft tumor mouse model, and also assessed the toxicity if any in normal mice. M. koenigii extract did not lead to any toxicity in mice. Analysis of extract revealed the presence of flavonoid compounds which act as proteasome inhibitors. Quercetin treatment led to the decrease in the cell viability and arrest of cells in G2/M phase. Quercetin, Apigenin, Kaempferol and Rutin; flavonoids present in the leaf extract, dose-dependently inhibited the endogenous 26S proteasome activity in MDA-MB-231 cells. Reduction in tumor growth was associated with a decrease in proteasomal enzyme activities in the treated groups. Increased caspase-3 activity and TUNEL-positive cells indicated enhanced apoptosis with Murraya leaf extract treatment. Decreased expression of angiogenic and anti-apoptotic gene markers is indicative of inhibition of angiogenesis and promotion of apoptosis in the leaf extract treated tumors.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Breast Neoplasms/drug therapy , Flavonoids/pharmacology , Murraya/chemistry , Plant Extracts/pharmacology , Proteasome Endopeptidase Complex/metabolism , Proteasome Inhibitors/pharmacology , Animals , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Breast Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Female , Flavonoids/chemistry , Flavonoids/isolation & purification , Humans , Mammary Neoplasms, Experimental/drug therapy , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Inbred BALB C , Mice, Nude , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Proteasome Inhibitors/chemistry , Proteasome Inhibitors/isolation & purification , Structure-Activity RelationshipABSTRACT
The vitamin D endocrine system is functional in the adipose tissue, as demonstrated in vitro, in cultured adipocytes, and in vivo in mutant mice that developed altered lipid metabolism and fat storage in the absence of either 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] or the vitamin D receptor. The aim of the present study was to examine the role of vitamin D and calcium on body adiposity in a diet-induced vitamin D deficient rat model. Vitamin D-deficient rats gained less weight and had lower amounts of visceral fat. Consistent with reduced adipose tissue mass, the vitamin D-deficient rats had low circulating levels of leptin, which reflects body fat stores. Expression of vitamin D and calcium sensing receptors, and that of genes involved in adipogenesis such as peroxisome proliferator-activated receptor, fatty acid synthase and leptin were significantly reduced in white adipose tissue of deficient rats compared to vitamin D-sufficient rats. Furthermore, the expression of uncoupling proteins (Ucp1 and Ucp2) was elevated in the white adipose tissue of the deficient rat indicative of higher energy expenditure, thereby leading to a lean phenotype. Expression of the p160 steroid receptor coactivator3 (SRC3), a key regulator of adipogenesis in white adipose tissue was decreased in vitamin D-deficient state. Interestingly, most of the changes observed in vitamin D deficient rats were corrected by calcium supplementation alone. Our data demonstrates that dietary vitamin D and calcium regulate adipose tissue function and metabolism.
Subject(s)
Adiposity/physiology , Ion Channels/genetics , Mitochondrial Proteins/genetics , Nuclear Receptor Coactivator 3/genetics , Vitamin D Deficiency/metabolism , Adipogenesis/genetics , Adiponectin/blood , Adipose Tissue/metabolism , Animals , Calcium/pharmacology , Diet , Fatty Acid Synthase, Type I/genetics , Gene Expression , Hypocalcemia/etiology , Hypocalcemia/genetics , Hypocalcemia/metabolism , Insulin/blood , Leptin/blood , Leptin/genetics , Lipids/blood , Liver/metabolism , Male , Nuclear Receptor Coactivator 3/metabolism , PPAR gamma/genetics , Rats, Sprague-Dawley , Uncoupling Protein 1 , Uncoupling Protein 2 , Vitamin D Deficiency/complications , Vitamin D Deficiency/geneticsABSTRACT
The enzyme 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) amplifies intracellular glucocorticoid action by converting inactive glucocorticoids to their active forms in vivo. Adipose-specific overexpression of 11ß-HSD1 induces metabolic syndrome in mice, whereas 11ß-HSD1 null mice are resistant to it. Dietary trans and saturated fatty acids (TFAs and SFAs) are involved in the development of metabolic syndrome, whereas polyunsaturated fatty acids (PUFA) offer protection against this. Here, we report the effects of chronic feeding of different diets containing vanaspati (TFA rich), palm oil (SFA rich) and sunflower oil (PUFA rich) at 10%level on 11ß-HSD1 gene expression in rat retroperitoneal adipose tissue. 11ß-HSD1 gene expression was significantly higher in TFA rich diet-fed rats compared to SFA rich diet-fed rats, which in turn was significantly higher than PUFA rich diet-fed rats. Similar trend was observed in the expression of CCAAT-enhancer binding protein-α (C/EBP-α), the main transcription factor required for the expression of 11ß-HSD1. We propose that TFAs and SFAs increase local amplification of glucocorticoid action in adipose tissue by upregulating 11ß-HSD1 by altering C/EBP-α-gene expression. The increased levels of glucocorticoids in adipose tissue may lead to development of obesity and insulin resistance, thereby increasing the risk of developing metabolic syndrome.
Subject(s)
11-beta-Hydroxysteroid Dehydrogenase Type 1/metabolism , Dietary Fats/administration & dosage , Fatty Acids/administration & dosage , Gene Expression Regulation, Enzymologic , Intra-Abdominal Fat/metabolism , 11-beta-Hydroxysteroid Dehydrogenase Type 1/genetics , Animals , Body Weight , CCAAT-Enhancer-Binding Protein-alpha/genetics , CCAAT-Enhancer-Binding Protein-alpha/metabolism , Dietary Fats/adverse effects , Dietary Fats/analysis , Fatty Acids/adverse effects , Fatty Acids/analysis , Fatty Acids, Unsaturated/administration & dosage , Fatty Acids, Unsaturated/analysis , Female , Insulin Resistance , Liver X Receptors , Metabolic Syndrome/epidemiology , Orphan Nuclear Receptors/genetics , Orphan Nuclear Receptors/metabolism , Palm Oil , Plant Oils/administration & dosage , Plant Oils/adverse effects , Plant Oils/chemistry , RNA, Messenger/metabolism , Rats , Rats, Inbred F344 , Reverse Transcriptase Polymerase Chain Reaction , Sunflower Oil , Trans Fatty Acids/administration & dosage , Trans Fatty Acids/adverse effects , Trans Fatty Acids/analysisABSTRACT
This study sought to determine whether effortful saliva swallows could be differentiated from habitual, noneffortful saliva swallows on the basis of swallow-related changes in neck circumference in humans. Gender differences in swallow-related neck circumference were examined as a secondary question. Twenty-seven healthy adults (14 females; mean age = 26.6 years, SD = 3.9 years) participated in two experimental runs (run duration = 10 min) during which they produced single trials of three visually cued tasks in random order: effortful saliva swallowing, saliva swallowing, and a control task involving repetitive apposition of the dominant thumb and index finger. Neck and ribcage circumference were simultaneously collected from the output of force transducers positioned around the neck and ribcage, respectively. The primary outcome variables were the positive and negative voltage peak amplitudes associated with changes in neck circumference during single-swallow trials. Effects of the swallowing task on positive and negative voltage peaks were examined with separate two-way analysis of variance procedures. Results indicated that both positive (F = 6.49, p < 0.05) and negative (F = 12.05, p Subject(s)
Biofeedback, Psychology/instrumentation
, Deglutition/physiology
, Physical Exertion/physiology
, Transducers, Pressure
, Adult
, Female
, Humans
, Male
, Muscle Contraction/physiology
, Neck
, Reproducibility of Results
, Saliva
, Sex Factors
, Young Adult