Low-ω3 Fatty Acid and Soy Protein Attenuate Alcohol-Induced Fatty Liver and Injury by Regulating the Opposing Lipid Oxidation and Lipogenic Signaling Pathways.

Chronic ethanol-induced downregulation of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) and upregulation of peroxisome proliferator-activated receptor gamma coactivator 1-beta (PGC1β) affect hepatic lipid oxidation and lipogenesis, respectively, leading to fatty liver injury. Low-ω3 fatty acid (Low-ω3FA) that primarily regulates PGC1α and soy protein (SP) that seems to have its major regulatory effect on PGC1β were evaluated for their protective effects against ethanol-induced hepatosteatosis in rats fed with Lieber-deCarli control or ethanol liquid diets with high or low ω3FA fish oil and soy protein. Low-ω3FA and SP opposed the actions of chronic ethanol by reducing serum and liver lipids with concomitant decreased fatty liver. They also prevented the downregulation of hepatic Sirtuin 1 (SIRT1) and PGC1α and their target fatty acid oxidation pathway genes and attenuated the upregulation of hepatic PGC1β and sterol regulatory element-binding protein 1c (SREBP1c) and their target lipogenic pathway genes via the phosphorylation of 5\u27 adenosine monophosphate-activated protein kinase (AMPK). Thus, these two novel modulators attenuate ethanol-induced hepatosteatosis and consequent liver injury potentially by regulating the two opposing lipid oxidation and lipogenic pathways

Protective Role of Dietary Curcumin in the Prevention of the Oxidative Stress Induced by Chronic Alcohol with respect to Hepatic Injury and Antiatherogenic Markers

Curcumin, an antioxidant compound found in Asian spices, was evaluated for its protective effects against ethanol-induced hepatosteatosis, liver injury, antiatherogenic markers, and antioxidant status in rats fed with Lieber-deCarli low menhaden (2.7% of total calories from ω-3 polyunsaturated fatty acids (PUFA)) and Lieber-deCarli high menhaden (13.8% of total calories from ω-3 PUFA) alcohol-liquid (5%) diets supplemented with or without curcumin (150 mg/kg/day) for 8 weeks. Treatment with curcumin protected against high ω-3 PUFA and ethanol-induced hepatosteatosis and increase in liver injury markers, alanine aminotransferase, and aspartate aminotransferase. Curcumin upregulated paraoxonase 1 (PON1) mRNA and caused significant increase in serum PON1 and homocysteine thiolactonase activities as compared to high ω-3 PUFA and ethanol group. Moreover, treatment with curcumin protected against ethanol-induced oxidative stress by increasing the antioxidant glutathione and decreasing the lipid peroxidation adduct 4-hydroxynonenal. These results strongly suggest that chronic ethanol in combination with high ω-3 PUFA exacerbated hepatosteatosis and liver injury and adversely decreases antiatherogenic markers due to increased oxidative stress and depletion of glutathione. Curcumin supplementation significantly prevented these deleterious actions of chronic ethanol and high ω-3 PUFA. Therefore, we conclude that curcumin may have therapeutic potential to protect against chronic alcohol-induced liver injury and atherosclerosis

Low- ω

Chronic ethanol-induced downregulation of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) and upregulation of peroxisome proliferator-activated receptor gamma coactivator 1-beta (PGC1β) affect hepatic lipid oxidation and lipogenesis, respectively, leading to fatty liver injury. Low-ω3 fatty acid (Low-ω3FA) that primarily regulates PGC1α and soy protein (SP) that seems to have its major regulatory effect on PGC1β were evaluated for their protective effects against ethanol-induced hepatosteatosis in rats fed with Lieber-deCarli control or ethanol liquid diets with high or low ω3FA fish oil and soy protein. Low-ω3FA and SP opposed the actions of chronic ethanol by reducing serum and liver lipids with concomitant decreased fatty liver. They also prevented the downregulation of hepatic Sirtuin 1 (SIRT1) and PGC1α and their target fatty acid oxidation pathway genes and attenuated the upregulation of hepatic PGC1β and sterol regulatory element-binding protein 1c (SREBP1c) and their target lipogenic pathway genes via the phosphorylation of 5′ adenosine monophosphate-activated protein kinase (AMPK). Thus, these two novel modulators attenuate ethanol-induced hepatosteatosis and consequent liver injury potentially by regulating the two opposing lipid oxidation and lipogenic pathways

Quercetin up-regulates paraoxonase 1 gene expression via sterol regulatory element binding protein 2 that translocates from the endoplasmic reticulum to the nucleus where it specifically interacts with sterol responsive element–like sequence in paraoxonase 1 promoter in HuH7 liver cells

We previously showed that quercetin expresses its antiatherogenic effects by up-regulating paraoxonase 1 (PON1) gene and high-density lipoprotein\u27s protective capacity against low-density lipoprotein oxidation. In an attempt to elucidate the mechanism of action of quercetin, we have now determined the effects of quercetin on PON1 gene expression, activity, protein level, nuclear mature sterol regulatory element binding protein 2 (SREBP2) level, and its translocation from the endoplasmic reticulum to nucleus and its interaction with PON1 promoter in human HuH7 liver cells using real-time reverse transcriptase polymerase chain reaction, spectrophotometry, immunoblot, confocal microscopy, and electrophoretic mobility shift assay techniques, respectively. Quercetin (20 μmol/L) treatment increased PON1 messenger RNA by 75% (P \u3c .02), with a concomitant 2-fold (P \u3c .05) increase in PON1 activity accompanied by 60% (P \u3c .01) increase in PON1 protein level. There was parallel to the 1.5- to 2.0-fold increase (P \u3c .05) in mature SREBP2 in the cell nuclei that was verified by increased immunolocalization of the mature SREBP2 (65-kd species) in the nuclei of quercetin-treated cells by confocal microscopy. Evaluation of the binding of biotin-labeled sterol responsive element (SRE)–like element of the PON1 promoter to the nuclear extract from the 24-hour quercetin (20 μmol/L)–treated HuH7 cells by electrophoretic mobility shift assay revealed that the SREBP2 specifically binds to the SRElike element that was abolished by prior incubation with anti-SREBP2 or significantly decreased by 200-fold molar excess of unlabeled SRElike sequence. Based on these results, we conclude that quercetin exhibits its antiatherogenic property by eliciting the translocation of the mature SREBP2 from endoplasmic reticulum to the nucleus, where it binds to SRE-like sequence in the PON1 promoter and up-regulates PON1 gene transcription and PON1 activity

Genetic engineering of cotton with a novel cry2AX1 gene to impart insect resistance against Helicoverpa armigera

Embryogenic calli of cotton (Coker310) were cocultivated with the Agrobacterium tumefaciens strain LBA4404 harbouring the codon-optimised, chimeric cry2AX1 gene consisting of sequences from cry2Aa and cry2Ac genes isolated from Indian strains of Bacillus thuringiensis. Forty-eight putative transgenic plants were regenerated, and PCR analysis of these plants revealed the presence of the cry2AX1 gene in 40 plants. Southern blot hybridisation analysis of selected transgenic plants confirmed stable T-DNA integration in the genome of transformed plants. The level of Cry2AX1 protein expression in PCR positive plants ranged from 4.9 to 187.5 ng g-1 of fresh tissue. A transgenic cotton event, TP31, expressing the cry2AX1 gene showed insecticidal activity of 56.66 per cent mortality against Helicoverpa armigera in detached leaf disc bioassay. These results indicate that the chimeric cry2AX1 gene expressed in transgenic cotton has insecticidal activity against H. armigera

Dietary Spermidine protects against Ethanol and Lipopolysaccharide induced Hepatic Oxidative Stress and Fibrosis

Alcohol liver disease (ALD) is a leading cause of death worldwide. The well-accepted two-hit model of ethanol and endotoxin, lipopolysaccharide (LPS) for ALD mimics the human disease by combining ethanol mediated oxidative stress caused by the metabolites of ethanol or endotoxin mediated increase in cytokines resulting in a deleterious positive feedback loop that propagates inflammation and fibrosis. In the first hit, ethanol is oxidized to acetaldehyde by alcohol dehydrogenase and cytochrome P4502E1 in hepatocytes, generating reactive oxygen species (ROS) within the cells. The second hit perpetuates liver injury and fibrosis as a result of endotoxin-induced activation of Kupffer cells resulting in oxidative stress and the activation of hepatic stellate cells (HSC), causing the up regulation of fibrogenic markers, platelet derived growth factor β-receptor (PDGFβR), α-smooth muscle actin (αSMA), collagen I (Col1) and fibronectin; and epigenetic repressor gene, methyl-CpG binding protein 2 (MeCP2). In contrast, the adipogenic gene, peroxisome proliferator-activated receptor γ (PPARγ) is suppressed leading to the transdifferentiation of HSC from quiescent to activated myofibroblastic phenotype, resulting in fibrosis. Spermidine, a naturally occurring polyamine found in soybean, mushrooms and whole grains is known for anti-aging properties and resistance to stress. In this study, the role of dietary spermidine in ethanol-LPS induced hepatic oxidative stress and fibrosis was investigated. Ethanol-LPS mediated liver injury was induced in C57BL/6 mice using chronic 5% ethanol fed in a liquid diet for 4 weeks plus a single dose of ethanol (5g/kg body wt, gavage) and LPS (2mg/kg body wt, ip), 6h prior to the experiment. Spermidine (5mg/kg body wt) was added to the liquid diet for 4 weeks. Oxidative stress was determined using ROS and glutathione (GSH) fluorescent assays and expression of superoxide dismutase (SOD). The above mentioned genes and their products were measured using RT-PCR and Western blot, respectively. The extent of fibrosis was determined by measuring the hydroxyproline content for collagen and Sirius Red stain. Spermidine protected against ethanol-LPS induced oxidative stress by decreasing the levels of ROS and the expression of SOD, and increasing the levels of the GSH. It also prevented the activation of HSC by suppressing the up regulated MeCP2, that coordinately reversed the down regulated PPARγ and the up regulated fibrogenic genes (αSMA, PDGFβR, Col1 and fibronectin). Spermidine also prevented fibrosis by decreasing hydroxyproline content and collagen fibers. These data suggest that spermidine may have potential as an anti-oxidant and anti-fibrotic agent for the prevention/treatment of ALD