HMG-CoA Reductase Gene Expression Research Articles

Expression levels of genes involved in lipogenesis and cholesterol synthesis in adenomyosis

Abstract Objectives Adenomyosis is a benign uterine disease that occurs with the invasion of the endometrial gland and stoma into the myometrium. The etiology and molecular pathology of adenomyosis are not yet fully understood. Tissue samples of patients diagnosed with adenomyosis and healthy endometrial tissues were investigated for the lipogenesis and cholesterol synthesis pathways. It was aimed to determine the difference between adenomyosis and healthy endometrial tissues in terms of lipid metabolism and to investigate the mechanism of adenomyosis in this context. Methods Formalin-fixed paraffin-embedded archival tissues were used in the current retrospective study. A total of 76 patient samples and 3 groups were used. Group 1: adenomyotic tissue (n=28), Group 2: eutopic endometrial tissue (n=30), and Control Group (n=18). In these groups, Sterol regulatory element binding protein 1 (SREBP1) molecule, fatty acid synthase (FASN), acetyl-CoA carboxylase (ACACA), ATP-citrate lyase (ACLY), HMG-CoA reductase (HMGCR), and HMG-CoA synthase (HMGCS) markers were evaluated by using RT-PCR method. Results Statistically significant differences (p<0.05) were found between the groups regarding expression levels of HMGCR, HMGCS, ACLY, ACACA, and SREBP1. HMGCR, HMGCS, ACLY, and SREBP1 gene expression levels between Group 1 and Group 2 and HMGCS, ACACA, ACLY, and SREBP1 gene expression levels between Group 1 and Control Group were determined as statistically different. A significant difference was detected only in HMGCR gene expression levels between Group 2 and the Control Group. Conclusions These results show that genes involved in lipid metabolism may be associated with the molecular pathogenesis of adenomyosis.

HMG-CoA reductase degrader, SR-12813, counteracts statin-induced upregulation of HMG-CoA reductase and augments the anticancer effect of atorvastatin

Statins are cholesterol-lowering drugs that have exhibited potential as cancer therapeutic agents. However, as some cancer cells are resistant to statins, broadening an anticancer spectrum of statins is desirable. The upregulated expression of the statin target enzyme, 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase (HMGCR), in statin-treated cancer cells is a well-known mechanism of statin resistance, which can be counteracted by the downregulation of HMGCR gene expression, or degradation of the HMGCR protein. However, the mechanism by which HMGCR degradation influences the anticancer effects of statins remain unreported. We tested the effect of the HMGCR degrader compound SR-12813 at a concentration that did not affect the growth of eight diverse tumor cell lines. Combined treatment with atorvastatin and a low concentration of SR-12813 led to lowering of increased HMGCR expression, and augmented the cytostatic effect of atorvastatin in both statin-resistant and -sensitive cancer cells compared with that of atorvastatin treatment alone. Dual-targeting of HMGCR using statins and SR-12813 (or similar compounds) could provide an improved anticancer therapeutic approach.

Rhamnetin decreases the expression of HMG-CoA reductase gene and increases LDL receptor in HepG2 cells

Context: Rhamnetin is a naturally occurring methylated derivative of quercetin. This flavonoid is abundant in Syzygium aromaticum, Coriandrum sativum Prunus cerasus, and Rhamnus spp. Aims: To evaluate the effects of rhamnetin on HMG-CoA reductase and low-density lipoprotein receptor (LDLR) gene and protein expressions in the HepG2 hepatoma cell line. Methods: The expression of HMG-CoA reductase and LDLR genes and proteins were studied in HepG2 liver cancer cell line by PCR, Western blot, and indirect ELISA, as well as their antioxidant activity. Results: Rhamnetin was non-toxic up to 200 μM on HepG2 at 24, 48, and 72 h. Rhamnetin (25 µM) upregulated LDLR gene expression by 1.66 folds compared to 3.12 folds exerted by the well-known hypocholesterolemic drug simvastatin. Rhamnetin (100 µM) increased the expression of LDLR protein at the cell membrane, while the other concentrations produced no significant change from the control (vehicle-treated). In HepG2 cell lysate, LDLR was increased by 50 µM of rhamnetin. Also, rhamnetin increased SOD activity significantly by 100.98, 86.28, and 100.98% by the concentrations 25, 50, and 100 µM, respectively. Using the same concentrations, rhamnetin reduced H2O2 levels by 50, 67, and 76.34%, respectively. Conclusions: This study demonstrated for the first time that rhamnetin reduced HMG-CoA reductase gene expression and increased LDLR in HepG2 cells.

Enterococcus faecium GEFA01 alleviates hypercholesterolemia by promoting reverse cholesterol transportation via modulating the gut microbiota-SCFA axis.

This study aimed to identify cholesterol-lowering commensal strains from healthy lean individuals and to evaluate the cholesterol-lowering capacity of Enterococcus faecium GEFA01 in mice fed a high-cholesterol and high-fat diet. E. faecium GEFA01 was isolated from the feces of a healthy lean individual in a selective basal salt medium supplemented with cholesterol. E. faecium GEFA01 exhibited a cholesterol removal rate (CRR) of 46.13% by coprecipitation, assimilation, and degradation of cholesterol. Moreover, E. faecium GEFA01 significantly decreased the body weight of mice and the levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), hepatic TC, triglycerides (TG), and LDL-C, and increased serum high-density lipoprotein cholesterol (HDL-C) levels in mice fed a high-cholesterol diet compared with the HCD group. We also observed that E. faecium GEFA01 significantly downregulated the gene expression of HMG-CoA reductase (Hmgcr), Srebp-1c, Fxr, Shp, and Fgf 15, upregulated the gene expression of low-density lipoprotein receptor (Ldlr), Abcg5/8, Abca1, cholesterol 7 alpha-hydroxylase (Cyp7a1), and Lxr in the liver of mice in relative to the HCD group, markedly increased the relative abundance of Lactobacillus, Akkermansia, Bifidobacterium, and Roseburia, and decreased the abundance of Helicobacter in the feces. Collectively, we confirmed that E. faecium GEFA01 exhibited cholesterol-lowering effects in mice fed a high-cholesterol diet, which was achieved through assimilation, coprecipitation, and degradation of cholesterol, and through modulation of the gut microbiota short-chain fatty acid (SCFA) axis that promoted reverse cholesterol transport and bile acid excretion. Our study demonstrated that E. faecium GEFA01 may be used as a probiotic candidate to lower cholesterol levels in the future.

Radix Angelica dahuricae extract ameliorates oestrogen deficiency-induced dyslipidaemia in ovariectomized (OVX) rats by modulating the gut microbiota and bile acid signalling

BackgroundRadix Angelica dahuricae (RAD), a well-known traditional Chinese medicine, displays a promising effect on alleviating lipid metabolism. However, the improvement of RAD on oestrogen deficiency-induced dyslipidaemia and the underlying mechanism are unclear. PurposeThe aim of this study was to study the effect of RAD on oestrogen deficiency-induced dyslipidaemia in ovariectomized (OVX) rats and investigate the involvement of the gut microbiota and bile acid signalling in the protective effects. MethodsBilateral ovariectomy was executed to establish an oestrogen deficiency model. Serum biochemical indexes, liver lipids, inflammatory cytokines and histomorphology were evaluated. Gut microbes were analysed via 16S rRNA sequencing. Faecal short-chain fatty acids (SCFAs) and serum bile acids were quantified by gas chromatography-flame ionization detection (GC-FID) and ultra-high-performance chromatography-tandem mass spectrometry (UPLC–MS/MS), respectively. The expression of genes related to bile acid synthesis, metabolism and enterohepatic circulation in the liver and caecum was measured by real-time PCR. ResultsThe results displayed that RAD administration markedly decreased body weight, TC and TG levels in the serum and liver, and hepatic steatosis and inflammation in OVX rats. RAD administration could significantly regulate the gut microbial composition, increasing the abundance of Lactobacillus, increasing the content of bile salt hydrolase (BSH), and reestablishing the SCFA profile and bile acid metabolism profile in OVX rats. RAD administration could increase the gene expression of HMG-CoA reductase (HMGCR) and cytochrome P450 7A1(CYP7A1) and regulate the gene expression of the related receptors as well as proteins in enterohepatic circulation. ConclusionsRAD alleviated oestrogen deficiency-induced dyslipidaemia in OVX rats. Modulation of the gut microbiota composition and bile acid signalling may be the underlying mechanism.

Effects of Momordica Charantia Silver Nanoparticles on the expressions of Genes Associated With Lipid Metabolism and Nephrotoxicity in Streptozotocin-Induced Rats

Hyperlipidemia and hyperglycemia have been implicated in diabetes mellitus (DM) leading to complications such as nephropathy. Medicinal plants like Mormodica charantia (MC) have been used in the treatment of DM over the years but little is known about their mechanisms of action. This study used biotechnology tools to investigate and compare the effects of M. charantia silver nanoparticles (MCSNPs) with M. charantia extract on expressions of genes linked with nephrotoxicity, lipid and glucose metabolisms using reverse-transcriptase polymerase chain reaction (RT-PCR) in streptozotocin-induced diabetic rats. The genes investigated include kidney injury molecule-1 (KIM-1), 3-hydroxyl, 3-methyl glutaryl_coA reductase (HMG-CoA reductase), peroxisome proliferator-activated receptor alpha and gamma (PPARα and PPARγ). Synthesis of MCSNPs was done using 1 mM concentration of aqueous silver nitrate solution at ratio 1:9 (v/v). Experimental rats were induced intraperitoneally with streptozotocin (65 mg/kg) and divided into six groups viz: diabetic control; normal control; silver nitrate (10 mg/kg); MCSNPs (50 mg/kg); Metformin (100 mg/kg) and M. charantia fraction (100 mg/kg). Sacrifice was done after 12 days of treatment and RT-PCR was then used to investigate gene expressions in liver and kidney tissues of the rats. The expression of HMG-CoA reductase gene was significantly upregulated (p<0.05) upon treatment with 50 mg/kg MCSNPs relative to the diabetic untreated group. M. charantia extracts and MCSNPs significantly upregulate (p<0.05) the expressions of PPAR-α and PPAR-γ compared to the diabetic control. Also, a significant (p<0.05) down-regulation of KIM-1 mRNA expression was observed in MCSNPs- treated group, relative to the diabetes untreated group. M. charantia silver nanoparticles could be a potent antidiabetic agent due to its potential to modulate genes associated with lipid metabolism and nephrotoxicity.
 Keywords: Medicinal plant; Diabetes Mellitus; Silver Nanoparticles; nephrotoxicity; gene expression

Annatto-Derived Tocotrienol Promotes Mineralization of MC3T3-E1 Cells by Enhancing BMP-2 Protein Expression via Inhibiting RhoA Activation and HMG-CoA Reductase Gene Expression.

PurposeAnnatto-derived tocotrienol (AnTT) has been shown to improve bone formation in animal models of osteoporosis and promote differentiation of pre-osteoblastic cells. However, the mechanism of action of AnTT in achieving these effects is unclear. This study aims to investigate the mechanism of action of AnTT on MC3T3-E1 pre-osteoblasts via the mevalonate pathway.MethodsMurine pre-osteoblastic cells, MC3T3-E1, were cultured with the density of 1 × 104 cells/mL and treated with 4 concentrations of AnTT (0.001–1 µg/mL). Expression of HMG-CoA reductase (HMGR) gene was carried out using qPCR after treatment with AnTT for 21 days. RhoA activation and bone morphogenetic protein-2 (BMP-2) were measured using immunoassay after 9 and 15 days of AnTT treatment. Lovastatin was used as the positive control. Mineralized nodules were detected using Von Kossa staining after 21 days of AnTT treatment.ResultsThe results showed that HMGR was up-regulated in the lovastatin group on day 9 and 21 compared to the control. Lovastatin also inhibited RhoA activation (day 9 and 15) and increased BMP-2 protein (day 15). On the other hand, AnTT at 0.001 μg/mL (day 3) and 0.1 μg/mL (day 21) significantly down-regulated HMGR gene expression compared to the control. On day 21, HMGR gene expression was significantly reduced in all groups compared to day 15. AnTT at 0.1 μg/mL significantly decreased RhoA activation on day 9 compared to the control. AnTT at 1 μg/mL significantly increased BMP-2 protein on day 15 compared to the control (P<0.05). Mineralized calcium nodules were more abundant in AnTT treated groups compared to the control on day 21.ConclusionAnTT suppresses the mevalonate pathway by downregulating HMGR gene expression and inhibiting RhoA activation, leading to increased BMP-2 protein in MC3T3-E1 cells. This explains the stimulating effects of AnTT on osteoblast mineralization.

Management of metabolic syndrome and reduction in body weight in type II diabetic mice by inhibiting glycosphingolipid synthesis

Metabolic syndrome is defined by hyperlipidemia and cardiovascular complications. We have examined whether inhibition of glycosphingolipid synthesis can interfere with metabolic syndrome in a male mouse model of type II diabetes (db/db). The db/db and control mice (C57/BL6) (n = 6) fed chow for 30 weeks received vehicle (5% Tween-80 in PBS; 100 μl), or a biopolymer-encapsulated D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (BPD) glycosphingolipid synthesis inhibitor daily via oral gavage for 6 weeks. Echocardiography revealed increased Ao-IMT in db/db mice compared to control. However, BPD decreased Ao-IMT, monohexosylceramide and dihexosylceramide, LDL, triglycerides, glucose, and raised HDL levels in db/db mice. This was due to increased gene expression of HMG-CoA reductase, LDLr, SREBP2, and bile acids: Cy7-a hydroxylase, LXR and FXR, lipoprotein lipase, VLDL receptor and PPAR. Treatment also increased the expression of superoxide dismutase-II to reduce the pro-oxidant status in these mice. We observed that decreased cholesterol levels correlated with decreased cholesterol sensing proteins e.g. NPC1 gene/protein expression and mammalian target of rapamycin (mTORC-1) and reduced body weight. Thus, glycosphingolipid synthesis inhibition is a novel approach to manage metabolic syndrome and reduce body weight in diabetic mice and with potential applications in humans.

Abstract P189: The Prorenin Receptor and its Soluble From Stimulate Hepatic Triglyceride and Cholesterol Metabolism in Male Mice

Non-alcoholic fatty liver disease is strongly associated with obesity and metabolic syndrome. Our laboratory previously demonstrated that mice lacking adipose prorenin receptor (PRR) have elevated hepatic triglycerides and cholesterol contents. In addition, lack of adipose PRR induced a counter regulatory increase of hepatic PRR and plasma soluble PRR (sPRR). Therefore, our study aimed to determine whether PRR and sPRR stimulated endogenous hepatic triglycerides and cholesterol synthesis. PRR floxed male mice fed a standard diet were injected with a single dose of an adeno-associated virus thyroxine-binding globulin Cre (KO, n=5) or saline (CTL, n=6). Body weight were measured weekly for 3 weeks. The deletion of PRR in liver did not change body weight curves. Hepatic levels of TG were significantly decreased in liver PRR KO mice compared with control mice (CTL, 615 ± 196 mg/g prot; KO, 114 ± 18 mg/g prot; P=0.047). PRR KO decreased significantly PPARγ gene expression suggesting that PRR positively regulates TG contents through PPARγ pathway. Surprisingly, similar to adipose PRR KO, liver PRR KO increased significantly plasma sPRR (CTL, 5942 ± 199 pg/ml; KO, 8903 ± 559 pg/ml; P=0.001) and hepatic total cholesterol levels (CTL, 40 ± 3 mg/g prot; KO, 102 ± 6 mg/g prot; P&lt;0.0001). This elevation was associated with an increase in SREBP-2 and HMGCoA-reductase gene expression. To determine whether sPRR stimulated cholesterol synthesis, HepG2 cells were treated with sPRR and sPRR was infused in C57BL/6J mice (n=7 to 8 mice/group). Results showed that sPRR significantly up-regulated hepatic SREPB2 mRNA expression in HepG2 cells (Veh, 1.00 ± 0.04; sPRR, 1.36 ± 0.08; P=0.009) and in the liver of mice (Veh, 1.01 ± 0.07; sPRR, 1.23 ± 0.07; P=0.04). In conclusion, our results demonstrated that hepatic PRR positively regulated TG contents through a PPARγ-dependent mechanism and that sPRR stimulated cholesterol synthesis via SREBP-2 pathway. Futures studies will investigate the mechanism by which PRR and sPRR modulate lipid metabolism.

Zinc oxide nanoparticles (ZnO NP) mediated regulation of bacosides biosynthesis and transcriptional correlation of HMG-CoA reductase gene in suspension culture of Bacopa monnieri

Bacopa monnieri (L.) Wettst. is a well documented nootropic plant, extensive known for alleviating symptoms of neurological disorder, along with other symptomatic relief. This property is attributed to the active phytocompounds, saponins (bacoside A) present in the plant. However, lack of stringent validation guidelines in most of the countries bring to the market, formulations differing in phytocompounds yield, thereby suggesting possible variation in therapeutic efficacy. The in-vitro suspension cultures of the Bacopa monnieri, provide an ease of scale-up, but regulating saponin yield is a stringent task. The aim of the study is to explore the effects of different concentrations (0, 0.25, 0.50, 0.75 and 1.0 ppm) of zinc oxide nanoparticles (ZnO NP) (24 nm in size), in regulating growth rate, bacoside yield and transcriptional profile of HMG CoA reductasegene in the suspension cells of Bacopa monnieri. Results showed a linear correlation between Bacoside A yield and ZnO NP concentrations with around 2 fold increase in total bacoside A concentration at 1 ppm. Also, ZnO NP supplemented suspension cells showed variation in the specific growth rate. Neuroprotective properties, analyzed using methanolic extracts of suspension cells again obtrude the extract of ZnO NP supplemented (0.75 ppm and 1 ppm) culture for better response in alleviating oxidative stress mediated damage to neuronal cells. ZnO NP supplemented system showed lower expression of HMG CoA reductasegene (the rate limiting step in bacoside A biosynthesis) but higher concentration of bacoside A, suggesting possible role of ZnO NP in isoprenoid pathway than MVA pathways.

7S protein is more effective than total soybean protein isolate in reducing plasma cholesterol

7S protein is one of four major storage proteins in soybean. The present study compared the relative cholesterol-lowering activity of the 7S protein with that of total soybean protein isolate (SPI). Forty-eight hamsters were divided into six groups fed non-cholesterol diet or one of five high-cholesterol diets containing 24.2% casein, 12.1–24.2% SPI, 12.1–24.2% 7S protein, respectively. Results showed the 7S protein reduced plasma total cholesterol (TC) by 20–41% while SPI reduced it by 9–16%. 7S protein increased the fecal excretion of neutral sterols by 43–116%, while SPI only had 18–68% increase. 7S protein increased the excretion of total bile acids by 208–367%, while SPI only increased it by 41–86%. 7S protein could not only down regulate gene expression of HMG-CoA reductase but also up regulate gene expression of CYP7A1. It was concluded that dietary 7S protein was more hypocholesterolemic than SPI, mediated by increasing the sterol excretion and decreasing the cholesterol synthesis.

Abstract 365: Inhibiting Hmg Coa Reductase Reduces Palmitate-Induced Inflammation in Adipocytes

Adipose tissue inflammation associates with insulin resistance and increased cardiovascular disease risk. We previously observed that 3T3-L1 adipocytes exposed to palmitate become inflamed and demonstrate increased plasma membrane cholesterol and lipid raft content. It is known that palmitate induces translocation of NAPH oxidase and toll-like receptor 4 into lipid rafts, increasing adipocyte inflammation. However, it is unclear (1) how palmitate alters plasma membrane cholesterol content; and (2) whether increased cholesterol content in the plasma membrane is related to adipocyte inflammation induced by palmitate exposure. We hypothesize that mechanisms involved in increasing plasma membrane cholesterol content after palmitate treatment could be related to cholesterol synthesis and/or ER stress, and that increased cholesterol in lipid rafts is essential for induction of inflammation in adipocytes. To test these hypotheses, differentiated murine 3T3-L1 adipocytes were exposed to palmitate for 24 hours, with and without pre-treatment with HMG-CoA reductase inhibitors (statins) or HDL. RT-PCR was used to evaluate gene expression of inflammation ( Saa3 , Ccl2 ), ER stress ( Bip , Chop ), and HMG-CoA reductase ( Hmgcr ). Cholera toxin subunit β staining and flow cytometry were used to evaluate plasma membrane lipid raft content. In differentiated adipocytes, palmitate-induced inflammation neither increased expression of ER stress genes nor HMG-CoA reductase gene expression. However, treatment with 3 different statins (simvastatin, lovastatin, atorvastatin) significantly reduced palmitate-induced adipocyte inflammation as indicated by decreased gene expression of Saa3 and Ccl2 ( P &lt;0.05). A similar effect was seen with pre-treatment with HDL. Lipid raft content induced by palmitate was decreased by HMG-CoA reductase inhibitors (difference in mean fluorescence intensity P &lt;0.05) and also by pre-treatment with HDL. These findings indicate that ER stress was not involved in increased plasma membrane cholesterol after palmitate-induced inflammation in adipocytes. However, regulating cholesterol content in lipid rafts plays an important role in adipocyte inflammation induced by palmitate.

A plant oxysterol, 28-homobrassinolide binds HMGCoA reductase catalytic cleft: stereoselective avidity affects enzyme function.

Understanding the influence of ubiquitously present plant steroids on mammalian cell biology is currently of interest. Feedback inhibition of HMGCoA reductase (HMGCR) catalytic activity in the transformation of HMG-CoA to mevalonate is a significant regulatory step in sterol biosynthetic pathway. To assess the role of dietary steroids in this biochemical transformation, the phytosteroid isoform 28-homobrassinolide (28-HB), 90% pure, obtained from Godrej Agrovet (India) was used to determine its effect on mammalian HMG-CoA reductase. Photometric assay of pure human and select rat tissue HMGCR post 28-HB oral feed, PCR-HMGCR gene expression, and in silico docking of 28-HB and HMGCoA on HMGCR protein template were carried out. Using an oral feed regimen of pure 28-HB, we noted a decrease of 16% in liver, 17.1% in kidney and 9.3% in testicular HMGCR enzyme activity, 25% in HMGCR gene expression and 44% in the activity of pure human HMGCR due to this plant oxysterol. In silico docking studies yielded binding metrics for 28-HB-HMGCR lower than for HMGCoA-HMGCR, indicating stronger binding of HMGCR by this ligand. 28-HB exerts differential effects on rat tissue HMGCR, down regulates liver HMGCR gene expression and significantly inhibits HMGCR activity.

Lipid Lowering Effects of Hydroalcoholic Extract of Anethum graveolens L. and Dill Tablet in High Cholesterol Fed Hamsters

Objective. This study was aimed to determine the effect of Anethum graveolens extract and Anethum graveolens (dill) tablet on lipid profile, liver enzymes, and gene expression and enzymatic activity of HMG-CoA reductase in high cholesterol fed hamsters. Materials and Methods. Golden Syrian male hamsters (130 ± 10 g) were randomly divided into 6 groups (n = 6) and received daily the following: group 1 received chow + 2% cholesterol + 0.5% cholic acid (HCD), groups 2 and 3 received HCD diet plus 100 and 200 mg/kg hydroalcoholic extract of dill, respectively, and groups 4 and 5 received HCD diet plus 100 and 200 mg/kg dill tablet, respectively. Group 6 received only chow. After 1 month feeding serum biochemical factors were determined. HMG-CoA reductase mRNA level was measured (real-time PCR) and its activity was determined spectrophotometrically. Results. Compared with hypercholesterolemic group 1, lipid profile, blood glucose, and liver enzymes significantly decreased in all dill tablet or dill extract treated groups (p < 0.05). The changes in HMG-CoA reductase gene expression level and enzyme activity significantly reduced in animals that received 200 mg/kg of extract or tablet. Conclusion. Dill extract and dill tablet showed potential hypocholesterolemic properties in hamsters by inhibition of HMG-CoA reductase activity.

Effects of Contaminated Fish Oil on Hepatic FAS and HMGCR Gene Expression and Liver Function Enzyme Activities

Concerns surrounding contamination with persistent organic pollutants (POPs) prompt caution in the consumption of fish and fish oil, despite their benefits on cardiovascular disease. The present study compared the effects of fish oil spiked with polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCs) to unmodified fish oil on hepatic gene expression of enzymes involved in lipid metabolism. Liver function enzyme levels were also examined. Thirty Sprague-Dawley rats were divided into three groups consuming a diet containing unmodified fish oil, fish oil spiked with POPs, or a control diet without fish oil. After 9 weeks of feeding, gene expression of fatty acid synthase (FAS) was significantly downregulated in both fish oil groups (P = 0.050) compared to control, while the unmodified fish oil group exhibited a tendency towards lower HMG-CoA reductase (HMGCR) transcription. This may explain the significantly lower levels of serum triglycerides in both fish oil groups (P = 0.002), while the unmodified fish oil group showed a tendency towards lower total cholesterol. Rats on the unmodified fish oil diet had significantly lower levels of lactate dehydrogenase (P = 0.040), while rats consuming diets with spiked fish oil exhibited significantly increased levels of alanine aminotransferase (P = 0.029). These findings point to differential genetic regulation from POP contamination and raise concerns surrounding liver damage during the consumption of spiked fish oils. Future studies should investigate the effects of a lengthened feeding time, varying contaminant doses, and interactions between contaminant mixtures and oils of varying composition. Supported by CHNR08-810, SDSU UGP and NUTR 302L class.

Hypolipidemic effect of black cumin (Nigella sativa) oil on adult layer birds

The present study was aimed to assess the effect of black cumin (Nigella sativa) oil on serum lipid profile and β-hydroxy, β-methylglutaryl coenzyme-A reductase (HMG CoA reductase) gene expression in adult layer birds. Thirty six weeks old White Leg Horn (WLH) layer birds were randomly divided into five groups consisting of 6 birds each. Their diets were supplemented with 0 (control), 1.0, 2.0, 3.0 and 4.0 g black cumin oil kg−1 feed, respectively. Blood samples were collected on day 0, 14, 28, 42 and 56 and study further continued for one more month without supplementing the diet with herbal preparations in order to observe the withdrawal effect. Gene expression studies of HMG CoA reductase were carried out in the liver tissue of the birds by real time quantitative PCR analysis on day 56. Serum total lipids, cholesterol, tri-glycerides, HDL, LDL and VLDL-cholesterol varied highly significantly (p 3> 2> 1> 0 g cumin oil kg−1 feed. The study revealed that the supplementation of black cumin oil @ 4 g kg−1 feed is most effective in reducing serum lipid profile and HMG CoA reductase gene expression after 56 days. These effects gradually reduced with time after the withdrawal of oil.

Effect of white wheat bread and white wheat bread added with bioactive compounds on hypercholesterolemic and steatotic mice fed a high-fat diet.

The effects of white wheat bread and white wheat bread added with a bioactive compound mixture (Cyclanthera pedata, Glycine max, Monascus-fermented red mold rice, Cynara scolymus and Medicago sativa) were examined on hypercholesterolemic and steatotic mice, divided into four groups: control diet (CTR), high-fat diet (HFD), high-fat diet with white wheat bread added with 1.5 g kg(-1) of mixture (HFD+AB) and high-fat diet with white wheat bread (HFD+B). Total serum cholesterol in the HFD+AB and HFD+B groups and hepatic triglycerides in the HFD+AB group decreased compared with the HFD group. Liver histology confirmed lower lipid drop accumulation in the HFD+AB group than in the HFD and HFD+B groups. HFD+AB caused a 7.0-fold increase and a 3.5-fold reduction in CYP7A1 and SREBP-1c gene expression respectively compared with the HFD group. Moreover, the HFD+B group showed a 2.2-, 8.4- and 1.5-fold increase in HMG CoA reductase, CYP7A1 and LDLr gene expression respectively compared with the HFD group. Both the white wheat bread and the added white wheat bread induced cholesterol reduction by increasing CYP7A1. Moreover, the added white wheat bread improved steatosis by decreasing SREBP-1c gene expression.

Olive oil and omega-3 polyunsaturated fatty acids suppress intestinal polyp growth by modulating the apoptotic process in ApcMin/+ mice

The promotion and progression of carcinogenesis are susceptible to nutritional interventions aimed at counteracting cancer development. Lipid metabolism is essential in the onset and progression of tumors and for cancer cell survival. In this study, we tested the effects of diets enriched with natural compounds, such as olive oil and salmon oil, in mice that spontaneously develop intestinal polyps (Apc(Min/+) mice). For this purpose, we evaluated polyp number and volume, intestinal mucosa proliferation/apoptosis, estrogen receptors (ERs) expression, fatty acid synthase and 3-hydroxy-3-methylglutaryl coenzyme A (HMGCoA) reductase gene expression and enzymatic activity. Compared with the standard diet, the salmon oil-enriched diet, containing a high percentage of omega-3 polyunsaturated fatty acids, and, to a lesser extent, olive oil-enriched diet reduced polyp number and volume through a reduction of proliferation and a marked proapoptotic effect. These biological effects were mediated by an inhibition of fatty acid synthase and HMGCoA reductase gene expression and activity and an increase of ERβ/ERα ratio. Our findings suggest that a proper dietary lifestyle could contribute to primary cancer prevention.

Lovastatin-enriched rice straw enhances biomass quality and suppresses ruminal methanogenesis.

The primary objective of this study was to test the hypothesis that solid state fermentation (SSF) of agro-biomass (using rice straw as model); besides, breaking down its lignocellulose content to improve its nutritive values also produces lovastatin which could be used to suppress methanogenesis in the rumen ecosystem. Fermented rice straw (FRS) containing lovastatin after fermentation with Aspergillus terreus was used as substrate for growth study of rumen microorganisms using in vitro gas production method. In the first experiment, the extract from the FRS (FRSE) which contained lovastatin was evaluated for its efficacy for reduction in methane (CH4) production, microbial population, and activity in the rumen fluid. FRSE reduced total gas and CH4 productions (P < 0.01). It also reduced (P < 0.01) total methanogens population and increased the cellulolytic bacteria including Ruminococcus albus, Fibrobacter succinogenes (P < 0.01), and Ruminococcus flavefaciens (P < 0.05). Similarly, FRS reduced total gas and CH4 productions, methanogens population, but increased in vitro dry mater digestibility compared to the non-fermented rice straw. Lovastatin in the FRSE and the FRS significantly increased the expression of HMG-CoA reductase gene that produces HMG-CoA reductase, a key enzyme for cell membrane production in methanogenic Archaea.

Lovastatin inAspergillus terreus: Fermented Rice Straw Extracts Interferes with Methane Production and Gene Expression inMethanobrevibacter smithii

Lovastatin, a natural byproduct of some fungi, is able to inhibit HMG-CoA (3-hydroxy-3methyl glutaryl CoA) reductase. This is a key enzyme involved in isoprenoid synthesis and essential for cell membrane formation in methanogenic Archaea. In this paper, experiments were designed to test the hypothesis that lovastatin secreted by Aspergillus terreus in fermented rice straw extracts (FRSE) can inhibit growth and CH4 production in Methanobrevibacter smithii (a test methanogen). By HPLC analysis, 75% of the total lovastatin in FRSE was in the active hydroxyacid form, and in vitro studies confirmed that this had a stronger effect in reducing both growth and CH4 production in M. smithii compared to commercial lovastatin. Transmission electron micrographs revealed distorted morphological divisions of lovastatin- and FRSE-treated M. smithii cells, supporting its role in blocking normal cell membrane synthesis. Real-time PCR confirmed that both commercial lovastatin and FRSE increased (P < 0.01) the expression of HMG-CoA reductase gene (hmg). In addition, expressions of other gene transcripts in M. smithii. with a key involvement in methanogenesis were also affected. Experimental confirmation that CH4 production is inhibited by lovastatin in A. terreus-fermented rice straw paves the way for its evaluation as a feed additive for mitigating CH4 production in ruminants.