Fatty Acid Synthetase Expression Research Articles

Effects of Dandelion Extract on Promoting Production Performance and Reducing Mammary Oxidative Stress in Dairy Cows Fed High-Concentrate Diet.

This study investigated the effects of rumen bypass dandelion extract on the lactation performance, immune index, and mammary oxidative stress of lactating dairy cows fed a high-concentrate diet. This study used a complete randomized block design, and initial milk production, somatic cell counts, and parities were set as block factors. Sixty Holstein cows with similar health conditions and lactating periods (70 ± 15 d) were divided into three groups with 20 replicates per group. The treatments included the LCD group (low-concentrate diet, concentrate-forage = 4:6), HCD group (high-concentrate group, concentrate-forage = 6:4), and DAE group (dandelion aqueous extract group, HCD group with 0.5% DAE). The experimental period was 35 d, and cows were fed three times in the morning, afternoon, and night with free access to water. The results showed the following: (1) Milk production in the HCD and DAE groups was significantly higher (p < 0.05) than that in the LCD group from WK4, and the milk quality differed during the experimental period. (2) The HCD group's pH values significantly differed (p < 0.01) from those of the LCD and DAE groups. (3) In WK2 and WK4 of the experimental period, the somatic cell counts of dairy cows in the HCD group were significantly higher (p < 0.05) than those in the DAE group. (4) The serum concentrations of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and protein carbonyl (PC) in the HCD group were significantly higher (p < 0.05) than those in the LCD group. The activity of catalase (CAT) in the LCD and DAE groups was stronger (p < 0.01) than that in the HCD group. (5) The correlation analysis revealed significantly positive correlations between the plasma LPS concentration and serum concentrations of 8-OHdG (p < 0.01), PC (p < 0.01), and malondialdehyde (MDA, p < 0.05) and significantly negative correlations (p < 0.01) between the plasma LPS concentration and activities of CAT and superoxide dismutase. (6) Compared with that in the HCD and DAE groups, the mRNA expression of α, β, and κ casein and acetyl CoA carboxylase in bovine mammary epithelial cells was significantly higher (p < 0.05) in the LCD group, and the mRNA expression of fatty acid synthetase and stearoyl CoA desaturase in the LCD group was significantly higher (p < 0.01) than that in the HCD group. (7) Compared with that in the LCD and HCD groups, the mRNA expression of Nrf2 was significantly higher (p < 0.01) in the DAE group, and the mRNA expression of cystine/glutamate transporter and NAD (P) H quinone oxidoreductase 1 in the DAE group was significantly higher (p < 0.05) than that in the HCD group. Overall, feeding a high-concentrate diet could increase the milk yield of dairy cows, but the milk quality, rumen homeostasis, and antioxidative capability were adversely affected. The supplementation of DAE in a high-concentrate diet enhanced antioxidative capability by activating the Nrf2 regulatory factor and improved rumen homeostasis and production performance.

Effects of Tetrahydrolipstatin on Glioblastoma in Mice: MRI-Based Morphologic and Texture Analysis Correlated with Histopathology and Immunochemistry Findings-A Pilot Study.

This study aimed to investigate the effects of tetrahydrolipstatin (orlistat) on heterotopic glioblastoma in mice by applying MRI and correlating the results with histopathology and immunochemistry. Human glioblastoma cells were injected subcutaneously into the groins of immunodeficient mice. After tumor growth of >150 mm3, the animals were assigned into a treatment group (n = 6), which received daily intraperitoneal injections of orlistat, and a control group (n = 7). MRI was performed at the time of randomization and before euthanizing the animals. Tumor volumes were calculated, and signal intensities were analyzed. The internal tumor structure was evaluated visually and with texture analysis. Western blotting and protein expression analysis were performed. At histology, all tumors showed high mitotic and proliferative activity (Ki67 ≥ 10%). Reduced fatty acid synthetase expression was measured in the orlistat group (p < 0.05). Based on the results of morphologic MRI-based analysis, tumor growth remained concentric in the control group and changed to eccentric in the treatment group (p < 0.05). The largest area under the receiver operating curve of the predictors derived from the texture analysis of T2w images was for wavelet transform parameters WavEnHL_s3 and WavEnLH_s4 at 0.96 and 1.00, respectively. Orlistat showed effects on heterotopically implanted glioblastoma multiforme in MRI studies of mice based on morphologic and texture analysis.

Targeting ZDHHC21/FASN axis for the treatment of diffuse large B-cell lymphoma

S-palmitoylation is essential for cancer development via regulating protein stability, function and subcellular location, yet the roles S-palmitoylation plays in diffuse large B-cell lymphoma (DLBCL) progression remain enigmatic. In this study, we uncovered a novel function of the palmitoyltransferase ZDHHC21 as a tumor suppressor in DLBCL and identified ZDHHC21 as a key regulator of fatty acid synthetase (FASN) S-palmitoylation for the first time. Specifically, ZDHHC21 was downregulated in DLBCL, and its expression level was associated with the clinical prognosis of patients with DLBCL. In vitro and in vivo experiments suggested that ZDHHC21 suppressed DLBCL cell proliferation. Mechanistically, ZDHHC21 interacted with FASN and mediated its palmitoylation at Cys1317, resulting in a decrease in FASN protein stability and fatty acid synthesis, consequently leading to the inhibition of DLBCL cell growth. Of note, an FDA-approved small-molecule compound lanatoside C interacted with ZDHHC21, increased ZDHHC21 protein stability and decreased FASN expression, which contributed to the suppression of DLBCL growth in vitro and in vivo. Our results demonstrate that ZDHHC21 strongly represses DLBCL cell proliferation by mediating FASN palmitoylation, and suggest that targeting ZDHHC21/FASN axis is a potential therapeutic strategy against DLBCL.

Periostin facilitates ovarian cancer recurrence by enhancing cancer stemness

The lethality of epithelial ovarian cancer (OC) is largely due to a high rate of recurrence and development of chemoresistance, which requires synergy between cancer cells and the tumor microenvironment (TME) and is thought to involve cancer stem cells. Our analysis of gene expression microarray data from paired primary and recurrent OC tissues revealed significantly elevated expression of the gene encoding periostin (POSTN) in recurrent OC compared to matched primary tumors (p = 0.015). Secreted POSTN plays a role in the extracellular matrix, facilitating epithelial cell migration and tissue regeneration. We therefore examined how elevated extracellular POSTN, as we found is present in recurrent OC, impacts OC cell functions and phenotypes, including stemness. OC cells cultured with conditioned media with high levels of periostin (CMPOSTNhigh) exhibited faster migration (p = 0.0044), enhanced invasiveness (p = 0.006), increased chemoresistance (p < 0.05), and decreased apoptosis as compared to the same cells cultured with control medium (CMCTL). Further, CMPOSTNhigh-cultured OC cells exhibited an elevated stem cell side population (p = 0.027) along with increased expression of cancer stem cell marker CD133 relative to CMCTL-cultured cells. POSTN-transfected 3T3-L1 cells that were used to generate CMPOSTNhigh had visibly enhanced intracellular and extracellular lipids, which was also linked to increased OC cell expression of fatty acid synthetase (FASN) that functions as a central regulator of lipid metabolism and plays a critical role in the growth and survival of tumors. Additionally, POSTN functions in the TME were linked to AKT pathway activities. The mean tumor volume in mice injected with CMPOSTNhigh-cultured OC cells was larger than that in mice injected with CMCTL-cultured OC cells (p = 0.0023). Taken together, these results show that elevated POSTN in the extracellular environment leads to more aggressive OC cell behavior and an increase in cancer stemness, suggesting that increased levels of stromal POSTN during OC recurrence contribute to more rapid disease progression and may be a novel therapeutic target. Furthermore, they also demonstrate the utility of having matched primary-recurrent OC tissues for analysis and support the need for better understanding of the molecular changes that occur with OC recurrence to develop ways to undermine those processes.

Beneficial Effect of Metformin, Quercetin, and Resveratrol Combination on High Glucose-Induced lipogenesis in HepG2 Cells

Objectives: It has been reported that Metformin (MET), Resveratrol (RSV), and Quercetin (QRS) possess anti-lipogenic effects. This study aimed to investigate the combined effects of these compounds on lipid accumulation in HepG2 cells treated with high glucose (HG).&#x0D; Methods: HepG2 cells were treated with HG (33 mM), and different concentrations of MET, QRS, and RSV. The cytotoxic effects of these compounds were determined by an MTT assay. Changes in total lipid content and triglyceride (TG) levels were measured using Oil Red O staining and a triglyceride assay kit, respectively. The expression of fatty acid synthetase (FAS) and sterol regulatory element-binding protein 1c (SREBP1c) was evaluated by quantitative real-time PCR.&#x0D; Results: MET at doses 1, 2, and 5 mM, QRS at doses 5, 10, and 20 µM, and RSV at doses 25 and 50 µM could decrease total lipid content and triglyceride levels in HepG2 cells. The EC50 (half maximal effective concentration) from Oil red O staining results were MET: 1.786 mM, QRS: 8.132 μM, and RSV: 10.9 μM. The combination of MET (mM), QRS (µM), and RSV (µM) at ratios of (2:20:50), (1:10:25), and (0.5:5:10), could reduce lipogenesis greater than that observed with each of the individual compounds of MET, QRS, or RSV or the double combinations of MET+QRS or MET+RSV. In addition, combined treatment of MET (0.5mM), QRS (5 µM), and RSV (10 µM) was able to decrease SREBP-1c and FAS genes expression in HG-treated cells.&#x0D; Conclusion: The combination of MET, QRS, and RSV could inhibit lipid accumulation in HepG2 cells by reducing total lipid content, triglyceride levels, and the expression of the genes involved in lipogenesis.

High-dose radiation-resistant lung cancer cells stored many functional lipid drops through JAK2/p-STAT3/FASN pathway.

The understanding of radiation resistance is still unclear. This study aims to explore the new mechanism of radiation resistance in lung cancer from the perspective of lipid metabolism. Oil red O was used to detect the amount of lipid droplets in high-dose radiation-resistant lung cancer cells (HDRR-LCCs) and the primary lung cancer cells. Western blot analysis was used to determine the protein expression levels of key molecules related to de novo fatty acid synthesis and fatty acid transport. Orlistat was used to inhibit the de novo fatty acid synthesis. The prediction of the transcriptional regulators of fatty acid synthetase (FASN) was analyzed by bioinformatics. AZD-1480 was used to inhibit the JAK2/STAT3 pathway to observe its effects on FASN and intracellular lipid droplets. The regulation of the transcription factor p-STAT3 on the FASN gene was verified by Chip-qPCR. Finally, we used the public data of lung cancer patients to analyze the correlation between FASN and LPL gene expression with the prognosis. There were more lipid drops in the HDRR-LCCs than in the primary lung cancer cells. HDRR-LCCs preferred de novo synthesis of fatty acids, and high expression of LPL homodimers indicated a high intake of extracellular fatty acids. The expression of FASN was increased in HDRR-LCCs compared with the primary lung cancer cells in a radiation-dose-dependent way, while LPL homodimers did not show such a trend. The lipid droplets, cell proliferation, and radiation resistance were decreased in HDRR-LCCs after orlistat treatment. Lipid droplets were significantly reduced, and the protein expression of FASN also decreased when using AZD-1480 to inhibit the JAK2/STAT3 pathway. The Chip-qPCR showed that p-STAT3 was the upstream regulator which binds to the promoter region of FASN. Survival analysis showed that high expression of the FASN gene was associated with a poor prognosis in lung cancer patients who received radiotherapy. Our studies discovered that lipids deposited in HDRR-LCCs were due to endogenous de novo fatty acids synthesis and exogenous lipids uptake. JAK2/p-TAT3/FASN could be used as promising targets for radiotherapy sensitization. Our study provided a new theoretical basis for studying the mechanism of radiation resistance in lung cancer.

Low Protein Diet Improves Meat Quality and Modulates the Composition of Gut Microbiota in Finishing Pigs.

This study investigated the effect of a low protein (LP) diet on growth performance, nitrogen emission, carcass traits, meat quality, and gut microbiota in finishing pigs. Fifty-four barrows (Duroc × Landrace × Yorkshire) were randomly assigned to three treatments with six replicates (pens) of three pigs each. The pigs were fed with either high protein (HP, 16% CP), medium protein (MP, 12% CP), and LP diets (10% CP), respectively. The LP diets did not influence the growth performance, but significantly decreased the plasma urea nitrogen contents and fecal nitrogen emission (P < 0.05). The LP diet significantly decreased the plasma contents of malondialdehyde (MDA) and increased the plasma glutathione (GSH) contents (P < 0.05). The LP diets significantly increased the backfat thickness at the first and last ribs, L* (lightness) value of meat color, and muscle fiber density in the longissimus dorsi (P < 0.05). The messenger RNA (mRNA) expression of fatty acid synthetase (FAS), peroxisome proliferator-activated receptor-gamma (PPARγ), leptin, and acetyl-CoA carboxylase (ACC) was significantly downregulated, while that of carnitine palmitoyltransferase 1 (CPT1) and myosin heavy chain (MYHC) IIx in the longissimus Dorsi muscle was significantly upregulated by LP diets (P < 0.05). The 16S sequencing analysis showed that the abundance of unidentified Bacteria at the phylum level, and Halanaerobium and Butyricicoccusat at the genus level in the colonic digesta were significantly decreased by LP diet (P < 0.05). The LP diet significantly decreased the observed species of α-diversity in both ileal and colonic microbiota (P < 0.05). Spearman correlation analysis identified a significant positive correlation between the abundance of the ileal genera Streptococcus and L* value at 24 and 48 h, and a significant negative correlation between unidentified_Ruminococcasceae in both ileum and colon with L* value at 24 h (P < 0.05). Collectively, the LP diet supplemented with lysine, methionine, threonine, and tryptophan could reduce the fecal nitrogen emission without affecting growth performance and improve meat quality by regulating the antioxidant capacity and gene expression involved in fat metabolism as well as modulating the gut microbiota composition in finishing pigs.

Hypoglycemic and Hypolipogenic Action of Acetic Acid and Monascus-Fermented Grain Vinegar: A Comparative Study.

Acetic acid has been proposed to improve lifestyle-related diseases, including hyperlipidemia and hyperglycemia. This study compared the hypoglycemic and hypolipogenic effects of acetic acid vinegar (AV, contains only 4% acetic acid) and Monascus-fermented grain vinegar (MV) containing various bioactive compounds in 3T3L1 cells and C57BL/KsJ-db/db mice (DB). The DB were divided randomly into three treatment groups containing nine mice each; DB-, AV-, and MV-groups were orally administered 1 mL/kg/day of distilled water, acetic acid vinegar, and Monascus vinegar, respectively, for 8 weeks. Exposure to AV and MV inhibited the adipogenic differentiation of 3T3L1 preadipocytes and lipid accumulation during differentiation. Oral administration of AV or MV to the mice resulted in a marked reduction in the body weight, liver weight, and hepatic triglyceride content compared to the control DB-group. Moreover, treatment with AV and MV clearly increased the expression of cyclic adenosine monophosphate (cAMP) and AMP-activated protein kinase (AMPK) and suppressed the expression of fatty acid synthetase in liver tissues of DB. Significantly, lower levels of fasting blood glucose, insulin, leptin, and the glycosylated hemoglobin (HbA1c) as well as higher levels of the skeletal muscle GLUT4 expression were obtained in the AV- or MV-groups than levels determined in the control DB-group (P < .05). Although MV has the potential to be a natural alternative treatment for obesity-associated type 2 diabetes, this study suggests that acetic acid is the central ingredient in MV responsible for the hypoglycemic and hypolipogenic effects in the DB mice.

Effects of maternal feed intake restriction on the blood parameters, fatty acid profile and lipogenetic genes expression of perirenal fat in offspring kids

Maternal nutritional restriction impacted lipogenic gene expression in adipose tissue of offspring, but the association of this programming with DNA methylation is not yet clear. Therefore, this experiment aimed to investigate the effects of maternal feed intake restriction on offsprings’ blood indexes, lipid metabolism of perirenal adipose tissue (PAT) and DNA methylation of lipogenic genes. Blood and PAT were collected from the fetuses (at d 100 of gestation) and kids (at d 90 after birth). Maternal undernutrition (at d 45 to d 100) decreases heart and kidney weight of kid goats after birth. Maternal undernutrition decreased the content of leptin in the umbilical cord blood of fetuses. The expression of uncoupling protein 1 (UCP1) was decreased in fetal PAT of the restriction group, but fatty acid synthetase (FASN) was increased. Protein abundance of UCP1 and FASN was down-regulated in the fetal PAT from the restriction group. Furthermore, only the methylation level of the CpG site (from −756 to −757 bp) in the promoter region of FASN gene was increased for the fetal PAT from the restriction group. These results revealed that maternal feed intake restriction influenced the leptin secretion, changed the expression of FASN and UCP1 gene of fetal PAT. This alteration was able to recover in the kids after removing intake restriction, but the growth performance and visceral organ mass were still impaired, suggesting abnormal fat metabolism may happen in the future.

Fatty acid synthetase expression in triple-negative breast cancer.

BackgroundTriple-negative breast cancer (TNBC) has a relatively poor prognosis. Research has identified potential metabolic targets, including fatty acid metabolism, in TNBC. The absence of effective target therapies for TNBC led to exploration of the role of fatty acid synthetase (FASN) as a potential target for TNBC therapy. Here, we analyzed the expression of FASN, a representative lipid metabolism–related protein, and investigated the association between FASN expression and Ki-67 and the programmed death ligand 1 (PD-L1) biomarkers in TNBC.MethodsImmunohistochemical expression of FASN was analyzed in 166 patients with TNBC. For analytical purposes, patients with 0–1+ FASN staining were grouped as low-grade FASN and patients with 2–3+ FASN staining as high-grade FASN.ResultsFASN expression was observed in 47.1% of TNBC patients. Low and high expression of FASN was identified in 75.9% and 24.1%, respectively, and no statistically significant difference was found in T category, N category, American Joint Committee on Cancer stage, or recurrence rate between the low and high-FASN expression groups. Ki-67 proliferation level was significantly different between the low and high-FASN expression groups. FASN expression was significantly related to Ki-67 as the level increased. There was no significant difference in PD-L1 positivity between the low- and high-FASN expression groups.ConclusionsWe identified FASN expression in 166 TNBC patients. The Ki-67 proliferation index was positively correlated with FASN level, indicating higher proliferation activity as FASN increases. However, there was no statistical association with PD-L1 SP142, the currently FDA-approved assay, or FASN expression level.

Gycyrrhizic acid alleviates atherosclerotic lesions in rats with diabetes mellitus.

Gycyrrhizic acid (GA), an inhibitor of high mobility group box 1 (HMGB1), inhibits inflammatory responses and is involved in the occurrence and development of several inflammation-related diseases. However, the role of GA in the atherosclerotic lesions caused by diabetes mellitus (DM) remains unknown. In the present study, Sprague Dawley rats were selected to desi=gn a diabetic atherosclerosis (AS) model. Rats from the DM-AS group were subsequently divided into DM-AS, DM-AS + GA (50 mg/kg) and DM-AS + GA (150 mg/kg) groups. Biochemical analyzers were used to measure levels of blood glucose, fasting insulin, total cholesterol, total triglyceride, low-density lipoprotein and high-density lipoprotein. The number of plaques was recorded after collection of thoracic aortas from the rats. The intimal thickness of arterial tissue was detected by hematoxylin and eosin staining. The expression levels of CD68 and α-smooth muscle actin (α-SMA) were detected by immunohistochemistry. The expression of tumor necrosis factor-α, interleukin (IL)-6 and IL-1β in the serum of the rats was detected by ELISA. The expression of fatty acid synthetase, sterol regulatory element binding protein 1C, HMGB1 and receptor for advanced glycation end products (RAGE) was detected by western blotting. Reverse transcription quantitative PCR was used to detect the mRNA expression of HMGB1 and RAGE. The results demonstrated that GA treatment could decrease the body weight, blood glucose level and biochemical parameters of AS DM rats in a dose-dependent manner. In addition, GA decreased the intimal thickness of carotid artery and the formation of plaque in rats with diabetic AS. Furthermore, GA inhibited macrophage activation and decreased α-SMA expression in vascular smooth muscle cells, and decreased the expression of proteins (FAS and SREBP-1c) and inflammatory factors. Taken together, the findings from the present study demonstrated that GA may have a therapeutic effect on DM-associated AS. This study provides a theoretical basis for the treatment of diabetic AS.

Proteomic response and molecular regulatory mechanisms of Bacillus cereus spores under ultrasound treatment

This study was aimed at providing new insights on the proteomic response of bacterial spores to ultrasound. Data-independent-acquisition method was used to quantify the proteome change of Bacillus cereus spores after ultrasound treatment (200 W). This study revealed that 2485 proteins were extracted from Bacillus cereus spores, most of them were related to metabolism. After ultrasound treatment, the expression of 340 proteins were significantly changed (the fold change ≥ 2 and p < 0.05), of which 207 proteins were significantly down-regulated. KEGG pathway analysis showed that differentially expressed proteins mainly distributed in metabolism pathway, cell process pathway and genetic information processing pathway after ultrasound treatment. Furthermore, this study analyzed the differentially expressed proteins in significant enrichment pathways. In particular, the expression of key proteins in the phosphorylation reaction of spores was significantly decreased after ultrasound treatment. Thus, ATP synthesis rate decreased and the phosphorylation reaction inhibited. Also, the decrease of the expression of key proteins related to the tricarboxylic acid cycle led to the decrease of nutrients metabolism of spores. Ultrasound treatment induced the down-regulation of fatty acid synthetase expression and promoted fatty acid metabolism at the same time. The content of fatty acids decreased in spores consequently.

Growth performance, meat quality and lipid metabolism in finishing lambs fed diets containing rumen-unprotected and rumen-protected betaine

This study investigated the influence of dietary supplementation of rumen-unprotected betaine (RUPB) and rumen-protected betaine (RPB) on growth performance, meat quality and fatty acid profile in intramuscular and subcutaneous fat of growing lambs. A total of 24 Hu male lambs were equally divided into three groups and fed a basal diet or basal diet supplemented with 1.6 g/kg of RUPB or 2.9 g/kg of RPB for 68 days. The water loss, redness of longissimus dorsi (LD) muscle and abdominal fat percentage were reduced in RUPB and RPB groups (p < .05). RPB supplementation decreased the shear force of LD muscle (p < .05). Polyunsaturated fatty acid (PUFA) content in LD muscle was decreased in RPB group, whereas n-3PUFA content was increased in RUPB group (p < .05). RUPB and RPB supplementation reduced the content of saturated fatty acid (SFA), and increased the contents of PUFA, monounsaturated fatty acid (MUFA), and n-6PUFA in subcutaneous fat (p < .05). Furthermore, dietary RUPB and RPB supplementation increased the mRNA expression of acetyl-CoA carboxylase (ACC) and glycerol-3-phosphate acyltransferase 1 (GPAT1) in LD muscle (p<.05); it increased the mRNA expression of fatty acid synthetase (FANS) and lipoprotein lipase (LPL) but reduced the mRNA expression of GPAT1 in subcutaneous fat (p < .05). In conclusion, either RUPB or RPB supplementation may improve the meat quality and slightly change the fat deposition and fatty acid composition of intramuscular and subcutaneous fat, and RUPB is recommended as a supplement for growing male lambs based on its lower cost. Highlights Dietary supplementation of both rumen-unprotected betaine and (RUPB) and rumen-protected betaine (RPB) improved meat quality and altered the fat deposition of growing male lambs. Both RUPB and RPB supplementation changed fatty acid composition in the longissimus dorsi muscle and subcutaneous adipose tissue. RUPB is recommended as a supplement for growing lambs based on its lower cost.

Effects of dietary lipid levels on survival, growth performance, and antioxidant ability of the early juvenile Scylla paramamosain

Five isonitrogenous diets (50% crude protein) with 7%, 9%, 11%, 13%, and 15% lipid levels (named as L7, L9, L11, L13, and L15, respectively) were designed to evaluate the effects of dietary lipid levels on survival, growth performance, body composition, and antioxidant ability of the early juvenile Scylla paramamosain. Each diet was fed to three replicates of 28 crabs with initial weight 42.23 ± 0.21 mg for 8 weeks. Crabs fed diet with lowest dietary lipid level (L7 group) obtained significantly lower survival than other crabs. The highest final body weight (FBW), weight gain (WG), specific growth rate (SGR), and molting frequency (MF) were observed in L11 group. Meanwhile, the highest mRNA expression of ecdysone receptor (EcR) also observed in L11 group. Besides, a positive correlation between body lipid contents and dietary lipid levels was observed in present study. But the mRNA expression of fatty acid synthetase (FAS) was relatively lower in L15 group. In the hepatopancreas, superoxide dismutase (SOD) activity significantly increased with increasing dietary lipid levels. On the contrary, the concentration of malondialdehyde (MDA) significantly decreased with elevating dietary lipid levels (except L15 group). Meanwhile, the lowest catalase (CAT) activity appeared in L7 group, but there was no statistical difference from other groups. Through the second order polynomial regression equation analysis of WG, the optimum dietary lipid level in the early juvenile S. paramamosain was 10.31%.

MicroRNA-212 targets SIRT2 to influence lipogenesis in bovine mammary epithelial cell line.

In this research paper we filter and verify miRNAs which may target silent information regulator homolog 2 (SIRT2) gene and then describe the mechanism whereby miRNA-212 might regulate lipogenic genes in mammary epithelial cell lines via targeting SIRT2. Bioinformatics analysis revealed that the bovine SIRT2 gene is regulated by three miRNAs: miR-212, miR-375 and miR-655. The three miRNAs were verified and screened by qRT-PCR, western blot, and luciferase multiplex verification techniques and only miR-212 was shown to have a targeting relationship with SIRT2. The results of co-transfecting miR-212 and silencing RNA (siRNA) showed that by targeting SIRT2, miR-212 can regulate the expression of fatty acid synthetase (FASN) and sterol regulatory element binding factor 1 (SREBP1) but not peroxisome proliferator-activated receptor gamma (PPARγ). Measurement of triglyceride (TAG) content showed that miR-212 increased the fat content of mammary epithelial cell lines. The study indicates that miR-212 could target and inhibit the expression of the SIRT2 gene to promote lipogenesis in mammary epithelial cell lines.

Antiadipogenesis of Oroxylum indicum (L.) Kurz Extract via PPARγ2 in 3T3-L1 Adipocytes.

Oroxylum indicum is regarded as a traditional food with medicinal properties and is used widely throughout Asia. It has previously been demonstrated that O. indicum extract (OIE) was able to suppress the differentiation of 3T3-L1 preadipocytes to adipocytes. However, the mechanism underlying the antiadipogenesis of this plant has not been fully investigated. The present study aimed to explore the impact of OIE at 50 to 200 μg mL−1 on the molecular mechanism involved in the antiadipogenic activity in 3T3-L1 cells at day 0 of their differentiation to adipocytes. The morphology and biochemistry of the cells on day 12 were investigated and compared to the relevant controls. Adiponectin was measured using enzyme-linked immunosorbent assay (ELISA). The mRNA expression of peroxisome proliferator-activated receptor-gamma 2 (PPARγ2), sterol regulatory element-binding proteins 1c (SREBP-1c), fatty acid synthetase (FAS), glucose transporter (GLUT4), and leptin in adipocytes was determined by real-time PCR. The results demonstrated that the OIE at 200 μg mL−1 exhibited strongest suppression on intracellular lipid accumulation. The levels of adiponectin were dramatically increased in the untreated adipocytes, whereas significantly decreased in the 200 μg mL−1 OIE-treated adipocytes (P < 0.05). Expression of the mRNAs revealed that OIE-treated adipocytes at 200 μg mL−1 significantly inhibited the expression of PPARγ2 and SREBP-1c and lowered the level of expression of GLUT4, FAS, and leptin compared to the control (P < 0.05). These findings suggest that OIE inhibits adipocyte differentiation along with the downregulation of PPARγ2, SREBP-1c, and GLUT4, leading to the decrease in the expression of FAS and adipokine (leptin and adiponectin). Thus, OIE might be developed for hyperlipidemia and obesity prevention.

Prenatal caffeine exposure increases the susceptibility to non-alcoholic fatty liver disease in female offspring rats via activation of GR-C/EBPα-SIRT1 pathway

This study aimed to evaluate female adult offspring induced by prenatal caffeine exposure (PCE) are susceptible to non-alcoholic fatty liver disease (NAFLD) and to explore the underlying programming mechanisms. Pregnant rats were intragastrically administered caffeine (30, 60, and 120 mg/kg.d) on gestational day (GD) 9-20. The female adult offspring were randomly divided into three groups: offspring without or with chronic stress during postnatal week (PW) 10–12 and PW28 offspring. Results showed that PW28 PCE female offspring had a higher hepatic triglyceride content and Kleiner scores, accompanied by elevated serum corticosterone levels. Moreover, the expression levels of hepatic glucocorticoid receptor (GR), CCAAT enhancer binding protein α (C/EBPα), fatty acid synthetase (FASN) and the transcription factor-sterol regulatory element binding protein 1c (SREBP1c) were increased, but SIRT1 expression was decreased. The fetal rats and PW12 offspring with chronic stress exhibited similar changes as PW28 offspring, accompanied by increased levels of H3K14ac and H3K27ac in the SREBP1c and FASN gene promoters. These effects were also observed by treating L02 cells with cortisol and were partially reversed by GR or C/EBPα siRNA or treatment with the SIRT1 agonist resveratrol. Taken together, PCE-induced high glucocorticoids levels enhanced histone modifications and expression of SREBP1c and FASN via activation of the GR-C/EBPα-SIRT1 pathway in utero. This enhanced female fetal hepatic triglyceride synthesis and continued throughout postnatal and adult life, increasing the susceptibility to adult NAFLD.

Chungkookjang with High Contents of Poly-γ-Glutamic Acid Improves Insulin Sensitizing Activity in Adipocytes and Neuronal Cells.

We hypothesized that soybeans fermented with Bacillus spp. for 48 h (chungkookjang) would be rich in poly-γ-glutamate (γ-PGA) and would have greater efficacy for improving insulin sensitivity and insulin secretion in 3T3-L1 adipocytes, min6 cells, and PC12 neuronal cells. We screened 20 different strains of B. subtillus and B. amyloliquefaciens spp. for γ-polyglutamate (PGA) production and their anti-diabetic and anti-dementia activities in cell-based studies. Chungkookjang made with two B. amyloliquefaciens spp. (BA730 and BA731) were selected to increase the isoflavonoid and γ-PGA. Insulin-stimulated glucose uptake was higher in 3T3-L1 adipocytes given both chungkookjang extracts than in the cells given vehicle (control). The ethanol extract of BA731 (BA731-E) increased the uptake the most. Triglyceride accumulation decreased in BA731-E and BA731-W and the accumulation increased in BA730-W and BA730-E. The mRNA expression of fatty acid synthetase and acetyl CoA carboxylase was much lower in BA731-E and BA731-W and it was higher in BA730-W than the control. BA730-E and BA730-W also increased peroxisome proliferator-activated receptor (PPAR)-γ activity. Glucose-stimulated insulin secretion increased with the high dosage of BA730-W and BA730-E in insulinoma cells, compared to the control. Insulin contents and cell survival in high glucose media were higher in cells with both BA731-E and BA730-E. Triglyceride deposition and TNF-α mRNA expression were lower in BA731 than the control. The high-dosage treatment of BA730-E and BA731-E increased differentiated neuronal cell survival after treating amyloid-β(25-35) compared to the control. Brain-derived neurotrophic factor and ciliary neurotrophic factor, indices of neuronal cell proliferation, were higher in BA730 and BA731 than in the control. Tau expression was also reduced in BA731 more than the control and it was a similar level of the normal-control. In conclusion, BA730 increased PPAR-γ activity and BA731 enhanced insulin sensitivity in the brain and periphery. BA730 and BA731 prevented and alleviated the symptoms of type 2 diabetes and Alzheimer’s disease with different pathways.

Mitochondrial damage in hippocampal neurons of rats with epileptic protein expression of Fas and caspase-3.

Epilepsy model in rats was established to observe the behavior and pathological changes, and to detect mitochondrial dysfunction, exploring its possible molecular mechanisms. The epileptic status of Sprague-Dawley (SD) rats was induced by intraperitoneal injection of lithium chloride, and the change of behavior was recorded. Electroencephalogram (EEG) was used to measure the abnormal discharge of neurons in rats. The brain tissue was fixed with polyformaldehyde and the paraffin sections were prepared, and the damage of the hippocampal neurons was observed with Nissl staining. Mitochondrial ATP and mitochondrial DNA were examined to assess mitochondrial dysfunction. Finally, qPCR and western blot analysis were used to detect mRNA and protein expression of fatty acid synthetase (Fas), Fas ligand (FasL) and caspase-3 in rat hippocampal neurons. The correlation between the mitochondrial dysfunction of rat hippocampal neurons and Fas and caspase-3 was analyzed. Compared with the normal group rats, the model group showed typical seizures, which were determined by the Racine attack score. EEG of the hippocampus of the model group was recorded in cluster in model group rats. Nissl staining showed a different degree of damage to the hippocampal neurons in the model group compared with normal rats. The mitochondrial ATP content and DNA content of rat hippocampal neurons in the model group were significantly lower than that of normal rats (P<0.01). The qPCR and western blot results showed that the mRNA and protein expression levels of Fas, FasL and caspase-3 were significantly increased in the hippocampus of rat model group (P<0.01). The expression level of Fas and caspase-3 in hippocampal tissues of rats was negatively correlated with mitochondrial DNA content. In conclusion, seizures cause damage of neuron mitochondria in rat hippocampus leading to death of hippocampal neurons, the mitochondrial damage of hippocampal neurons in epileptic rats was closely related to the expression of Fas and caspase-3.

Transcriptional regulation of milk lipid synthesis by exogenous C16:0 and C18 fatty acids in bovine mammary epithelial cells

The objective of this study was to examine the effects of removing one fatty acid from a combination of long-chain fatty acids (LCFA) on milk lipogenesis in bovine mammary epithelial cells. The incubation concentration of LCFA was determined, and 100 μmol L−1 of C16:0, 5 μmol L−1 of C18:0, 100 μmol L−1 of cis-9 C18:1, 25 μmol L−1 of n-6 C18:2, and 1.2 μmol L−1 of n-3 C18:3 were used in the study. Treatments were C16:0, C18:0, C18:1, C18:2, and C18:3 combinations as control; control absent of C16:0 as A-C16:0; control absent of C18:0 as A-C18:0; control absent of C18:1 as A-C18:1; control absent of C18:2 as A-C18:2; control absent of C18:3 as A-C18:3. Results showed that compared with control, fatty acid synthetase expression was reduced by A-C18:0 and A-C18:1. Palmitic acid decreased expression of lipoprotein lipase. Compared with control, the expression of stearoyl-coenzyme A desaturase-1 and cluster of differentiation 36 was reduced by all treatments. Peroxisome proliferator-activated receptor-α expression was down-regulated by A-C16:0, A-C18:0, A-C18:1, and A-C18:2. Sterol regulatory element binding factor-1 was decreased when treated with A-C18:0, A-C18:1, and A-C18:2. Cells lack of 18-carbon fatty acid synthesized lower amount of intracellular triglyceride compared with control.