ABSTRACT
The peroxisome proliferator-activated receptor alpha (PPARalpha) has been implicated as a key control of fatty acid catabolism during the cellular fasting. However, little is known regarding changes of individual fatty acids in hepatic triacylglycerol (TG) and phospholipid (PL) as a result of starvation. In the present work, the effects of 72 h fasting on hepatic TG and PL fatty acid profiles in PPARalpha-null (KO) mice and their wild-type (WT) counterparts were investigated. Our results indicated that mice deficient in PPARalpha displayed hepatomegaly and hypoketonemia following 72 h starvation. Histochemical analyses revealed that severe fatty infiltration was observed in the livers of KO mice under fasted conditions. Furthermore, 72 h fasting resulted in a 2.8-fold higher accumulation of hepatic TG in KO mice than in WT mice fasted for the same length of time. Surprisingly, the total hepatic PL contents in fasted KO mice decreased by 45%, but no significant change in hepatic PL content was observed in WT mice following starvation. Gas chromatographic analysis indicated that KO mice were deprived of arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids during fasting. Taken together, these results show that PPARalpha plays an important role in regulation of fatty acid metabolism as well as phospholipid homeostasis during energy deprivation.
Subject(s)
Food Deprivation , PPAR alpha/physiology , Phospholipids/metabolism , Triglycerides/metabolism , 3-Hydroxybutyric Acid/blood , Animals , Arachidonic Acid/metabolism , Arachidonic Acids/metabolism , Azo Compounds/pharmacology , Blotting, Northern , Carbohydrates/chemistry , Cholesterol/blood , Chromatography, Gas , Coloring Agents/pharmacology , Docosahexaenoic Acids/metabolism , Fasting , Fatty Acids/metabolism , Glycogen/chemistry , Homeostasis , Lipid Metabolism , Liver/metabolism , Male , Mice , Mice, Knockout , Naphthalenes , PPAR alpha/metabolism , Periodic Acid/pharmacology , Phospholipids/chemistry , RNA/metabolism , Time Factors , Triglycerides/bloodABSTRACT
Polycyclic aromatic hydrocarbons (PAHs) are established cancer initiators that can be found in our food and environment. Some dietary plant polyphenols are strong inhibitors to PAH-induced mutagenesis, whereas others may not be as effective. To identify the chemopreventive compounds from a huge volume of dietary components, the development of an efficient screening method is required. In this study, a xenobiotic response element (XRE)-luciferase reporter plasmid was constructed to screen for some potential chemopreventive agents in tea against PAH-induced DNA damage. Tea is one of the most consumed beverages worldwide, and its beneficial effects on health have been documented. Previous studies have claimed that tea polyphenols could be protective against various cancers, and the rich database can be a source for comparison. Among the green and black tea polyphenols, the XRE-luciferase reporter assays suggested that only epigallocatechin gallate (EGCG) was effective in reducing XRE-driven luciferase assay in MCF-7 cells at the concentrations tested. Further study indicated EGCG could reduce CYP1A1 and CYP1B1 mRNA abundances and decrease the DMBA-DNA lesions. The results of DNA covalent binding of all tea polyphenols tested were consistent with the XRE-reporter assays. This study illustrated that the XRE-reporter assay was a viable screening test for dietary chemopreventive agents against PAH-initiated breast mutagenesis. It has the advantages of shorter sample processing time and producing no radioactive waste over directly measuring the CYP1A1/1B1 expressions, DNA lesion, or gel mobility shift assay.
Subject(s)
Breast Neoplasms/prevention & control , Flavonoids/therapeutic use , Luciferases , Phenols/therapeutic use , Polycyclic Aromatic Hydrocarbons/toxicity , Tea , Xenobiotics , Analysis of Variance , Animals , Breast Neoplasms/genetics , Cell Survival/drug effects , DNA Damage/drug effects , Drug Evaluation, Preclinical , Genes, Reporter/physiology , Polyphenols , Rats , Reverse Transcriptase Polymerase Chain Reaction , Tumor Cells, Cultured/drug effectsABSTRACT
Genistein is a soy isoflavone that has attracted much attention for its chemopreventive properties. Baicalein is a flavone found in Scutellaria species, and its structure is analogous to that of genistein. We examined the two flavones' antiestrogenicity by using a transient transfection model. Our results indicated that baicalein could suppress 17beta-estradiol-induced transactivation in cells expressing estrogen receptor alpha, whereas no antagonistic activity was seen in genistein. Cell death assay and flow cytometry analysis demonstrated that baicalein was a stronger apoptosis inducing agent than genistein. Considering all the criteria tested, baicalein could be a better chemopreventive agent than genistein.
Subject(s)
Antioxidants/pharmacology , Apoptosis/drug effects , Breast Neoplasms/metabolism , Carcinoma, Hepatocellular/metabolism , Flavanones , Flavonoids/pharmacology , Genistein/pharmacology , Receptors, Estrogen/metabolism , Antineoplastic Agents/pharmacology , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Cell Division , Estrogen Receptor alpha , Female , Flow Cytometry , Humans , Luciferases/metabolism , Receptors, Estrogen/genetics , Response Elements , Topoisomerase II Inhibitors , Transcriptional Activation/drug effects , Transfection , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/metabolismABSTRACT
Flavonoids are phenolic compounds isolated from plants, and several of them like genistein and quercetin, have been documented to be effective in preventing cancer. Baicalein, a flavonoid extracted from the root of Scutellaria species, is widely used as a health supplement and herbal medicine in Asian countries. In this study, the chemopreventive effect of baicalein on 7,12-dimethylbenz[a]anthracene (DMBA)-induced DNA damage was evaluated in an established cell culture model. In a preliminary screening, baicalein was identified to be a strong inhibitor to EROD activities induced by DMBA in MCF-7 cells. Subsequent enzyme kinetic analysis revealed that baicalein was a competitive inhibitor to EROD, and CYP1A1 and CYP1B1 gene expressions were also determined. Baicalein could reduce the CYP1A1/1B1 mRNA expressions induced by DMBA, and the mRNA abundance of CYP1A1 appeared to be more responsive than that of CYP1B1. A XRE-luciferase gene reporter assay indicated that AhR transactivation was suppressed. Since CYP1A1/1B1 were responsible for the biotransformation of polycyclic aromatic hydrocarbons, baicalein also demonstrated its ability to reduce DMBA-DNA adduct formation in MCF-7 cells. This study suggested that the natural occurring baicalein could be an agent preventing carcinogen-DNA adduct formation.
