UBE2S activates NF-κB signaling by binding with IκBα and promotes metastasis of lung adenocarcinoma cells.

PURPOSE: Nuclear factor (NF)-κB signaling in cancer cells has been reported to be involved in tumorigenesis. Phosphorylation and degradation of inhibitor of NF-κBα (IκBα) is a canonical pathway of NF-κB signaling. Here, we aimed to identify and characterize noncanonical activation of NF-κB signaling by ubiquitin-conjugating enzyme E2S (UBE2S) in lung adenocarcinoma cells. METHODS: TCGA and the Human Atlas Protein Database were used to analyze the survival rate of lung adenocarcinoma patients in conjunction with UBE2S expression. In addition, PC9, H460, H441 and A549 lung adenocarcinoma cells were used in this study. PC9 and H460 cells were selected for further analysis because they expressed different UBE2S protein levels. Specific IKK inhibitors, PS1145 and SC514, were used to assess IκBα phosphorylation. Western blot analysis was used to assess protein levels in PC9 and H460 cells. A scratch wound-healing assay was used to analyze the migrative abilities of PC9 and H460 cells. Overexpression and knockdown of UBE2S in H460 and PC9 cells were used to analyze their effects on downstream protein levels. Immunoprecipitation, immunofluorescent staining, glutathione S transferase (GST) pull-down and in vitro binding assays were used to analyze the interaction between UBE2S and IκBα. A luciferase assay was used to analyze activation of NF-κB signaling regulated by UBE2S. An in vivo zebrafish xenograft model was used to assess metastasis of PC9 cells regulated by UBE2S. RESULTS: We found that UBE2S expression in lung adenocarcinoma patients was negatively related to survival rate. The protein level of UBE2S was higher in PC9 cells than in H460 cells, which was opposite to that observed for IκBα. PC9 cells showed a higher UBE2S expression and migrative ability than H460 cells. Phosphorylation of IκBα was not changed by treatment with the IKK-specific inhibitors PS1145 and SC514 in PC9 and H460 cells. Overexpression and knockdown of UBE2S in H460 and PC9 cells revealed that the protein levels of IκBα were inversely regulated. Immunoprecipitation, immunofluorescent staining, GST pull-down and in vitro binding assays revealed direct binding of UBE2S with IκBα. Nuclear P65 protein levels and luciferase assays showed that NF-κB signaling was regulated by UBE2S. The expression of epithelial-to-mesenchymal (EMT) markers and the migrative ability of lung adenocarcinoma cells were also regulated by UBE2S. A zebrafish xenograft tumor model showed a reduction in the metastasis of PC9 cells that was induced by UBE2S knockdown. CONCLUSIONS: Higher UBE2S expression in lung adenocarcinomas may lead to increased binding with IκBα to activate NF-κB signaling and promote adenocarcinoma cell metastasis. UBE2S may serve as a potential therapeutic target for lung adenocarcinomas.

Effects of rice bran oil on the blood lipids profiles and insulin resistance in type 2 diabetes patients.

The aim of this study was to investigate the influence of rice bran oil consumption on plasma lipids and insulin resistance in patients with type 2 diabetes. Thirty-five patients with type 2 diabetes were randomly assigned to a placebo group or a rice bran oil group. The placebo group consumed 250 mL soybean oil-modified milk (18 g soybean oil) daily for 5 weeks, and the rice bran oil group consumed 250 mL rice bran oil modified milk (18 g rice bran oil) daily for 5 weeks. At week 0 and week 5, anthropometric measurements, hematology tests, and an oral-glucose-tolerance test were conducted. The results showed that the homeostasis model assessment index of insulin resistance, the area under the curve for postprandial serum insulin, and serum low-density-lipoprotein cholesterol concentrations increased significantly in the placebo group. In the rice bran oil group, fasting and 2-h postprandial blood glucose concentrations and the area under the curve for postprandial plasma glucose increased significantly; however, total serum cholesterol and low-density-lipoprotein cholesterol concentrations decreased significantly. However, the homeostasis model assessment index of insulin resistance was not significantly different. Consumption of 18 g rice bran oil modified milk daily for 5 weeks significantly decreased total serum cholesterol concentrations and tended to decrease low-density-lipoprotein cholesterol concentrations in patients with type 2 diabetes. However, no significant influence on insulin resistance was observed.

Gamma-oryzanol ameliorates insulin resistance and hyperlipidemia in rats with streptozotocin/nicotinamide-induced type 2 diabetes.

Gamma-oryzanol is a component of rice bran oil (RBO) with purported health benefits. This study evaluated the effects of gamma-oryzanol on insulin resistance and lipid metabolism in Wistar rats with type 2 diabetes (T2DM). The rats were divided into three groups and consumed one of the following diets for 5 weeks: 15 % soybean oil (control group); 15 % palm oil (PO); and 15 % PO with the addition of 5.25 g gamma-oryzanol (POO). The results showed that PO markedly increased plasma low-density-lipoprotein cholesterol, plasma triglycerides, and hepatic triglyceride levels, but did not reduce the area under the curve for glucose and insulin significantly, compared with the control group. Adding gamma-oryzanol to PO improved the negative influence of PO on lipid metabolism in T2DM rats. In addition, gamma-oryzanol tended to increase insulin sensitivity in T2DM rats compared to control and PO groups. Longer-term studies are needed to evaluate these effects further.

Ameliorative effects of stabilized rice bran on type 2 diabetes patients.

BACKGROUND/AIM: While it has been demonstrated that rice bran might lower the cholesterol level in hypercholesterolemic individuals, its effects on the levels of adiponectin and glycated hemoglobin (HbA1c) in patients with type 2 diabetes mellitus remain unknown. METHODS: Twenty-eight volunteers with type 2 diabetes were randomly divided into 2 groups, one of which received a dietary supplement of 20 g of stabilized rice bran and the other placebo once daily for 12 weeks. Parameters such as the level of HbA1c, glucose, insulin, homeostasis model assessment for estimation of relative insulin resistance, high-density and low-density lipoprotein (LDL) cholesterol and adiponectin were evaluated. RESULTS: At the end of the study period, postprandial glucose and the area under the glucose curve of the rice bran group were significantly lower than baseline levels by 14.4 and 15.7%, respectively. Compared to baseline, the HbA1c values in the rice bran group were also significantly lower. Serum total cholesterol and LDL cholesterol concentrations in the rice bran group were 9.2 and 13.7% lower, respectively, than in the placebo group. The plasma free fatty acid and adiponectin concentrations were 20% lower and 40% higher in the rice bran group compared to the placebo group. CONCLUSIONS: This study demonstrated that stabilized rice bran can lower the level of HbA1c and blood lipids and increase blood adiponectin concentrations in type 2 diabetic subjects. In light of this, we conclude that stabilized rice bran may represent an important functional nutrient to ameliorate lipid and glycemic anomalies in type 2 diabetic subjects.

A rice bran oil diet improves lipid abnormalities and suppress hyperinsulinemic responses in rats with streptozotocin/nicotinamide-induced type 2 diabetes.

The aim of this study was to determine the effects of rice bran oil (RBO) on lipid metabolism and insulin resistance in rats with streptozotocin/nicotinamide-induced type 2 diabetes mellitus (T2DM). Rats were divided into two groups: the control group (15% soybean oil, contains 0 g gamma-oryzanol and 0 g gamma-tocotrienol/150 g oil for 5 weeks) and the RBO group (15% RBO, contains 5.25 g gamma-oryzanol and 0.9 g gamma-tocotrienol/150 g oil for 5 weeks). Compared with the control group, the RBO group had a lower plasma nonesterified fatty acid concentration, ratio of total to high-density-lipoprotein cholesterol, hepatic cholesterol concentration, and area under the curve for insulin. The RBO group had a higher high-density-lipoprotein cholesterol concentration and greater excretion of fecal neutral sterols and bile acid than did the control group. RBO may improve lipid abnormalities, reduce the atherogenic index, and suppress the hyperinsulinemic response in rats with streptozotocin/nicotinamide-induced T2DM. In addition, RBO can lead to increased fecal neutral sterol and bile acid excretion.

Linoleic acid promotes mitochondrial biogenesis and maintains mitochondrial structure for prevention of streptozotocin damage in RIN-m5F cells.

Linoleic acid (LA) improves insulin resistance and prevents diabetes. To investigate whether linoleic acid could protect against streptozotocin (STZ)-induced cell death, rat RIN-m5F cells were exposed to STZ. SL and SO groups consisted of cells treated with STZ and then LA or oleic acid (OA) respectively. STZ treatment decreased the mitochondrial membrane potential in the STZ, SO, and SL groups. Cells of the SL group had more intact mitochondria. Increased mRNA expression of mitochondrial DNA (mtDNA) and nuclear DNA (nDNA), as well as of the mitochondrial biogenesis regulators peroxisome proliferator activated receptor gamma coactivator-1alpha (PGC-1alpha), and mitochondrial transcription factor A (Tfam), were found in the LA group. The insulin content was significantly decreased in all three groups. These results suggest that the effects of LA on cell viability after STZ damage occur through maintenance of mitochondrial structure and increased mitochondrial biogenesis.

beta-Carotene and canthaxanthin alter the pro-oxidation and antioxidation balance in rats fed a high-cholesterol and high-fat diet.

This study investigated the effects of beta-carotene and canthaxanthin on lipid peroxidation and antioxidative enzyme activities in rats fed a high-cholesterol, high-fat diet. Wistar rats were divided into six groups. Negative control group (group NC) received a high-fat (150 g/kg) diet; cholesterol control group (group CC) received a high-cholesterol (10 g/kg), high-fat diet. The other four groups were fed a high-cholesterol, high-fat diet supplemented with crystal beta-carotene (group BC), beta-carotene beadlet (group BB), canthaxanthin beadlet (group CX) or alpha-tocopherol (group AT). Blood and livers were collected for analysis after 6 weeks of feeding. Group BB had significantly lower hepatic thiobarbituric acid reactive substance (TBARS) and conjugated diene concentrations, whereas group CX had a significantly lower plasma TBARS concentration than did group CC. In erythrocytes, glutathione peroxidase activities were significantly greater in groups BC, BB and CX than in group CC. Moreover, compared with group CC, catalase activities were significantly greater in groups BB and CX, and superoxide dismutase (SOD) activity was significantly greater in group BB. In livers, SOD activities were significantly greater in groups BC, BB and CX, and glutathione reductase activities were significantly greater in groups BB and CX than in group CC. Compared with group CC, hepatic retinol and alpha-tocopherol concentrations were significantly greater in groups BC, BB and CX, whereas plasma and hepatic cholesterol concentrations were significantly lower in group BC. These findings suggest that beta-carotene and canthaxanthin altered the pro-oxidation and antioxidation balance and suppressed cholesterol-induced oxidative stress via modulation of antioxidant system and cholesterol metabolism.

Fruits and stir-fried vegetables increase plasma carotenoids in young adults.

We examined the plasma levels of carotenoids in young adults after a dietary intervention composed of increased intakes of fruits and stir-fried vegetables from a Taiwanese mixed diet. Thirty-four apparently healthy, non-smoking subjects who normally ingested less than two and a half servings of fruits and vegetables daily were selected for the study. Meals changed were lunch and dinner on weekdays for a period of 4 weeks. The test meal consisted of three servings of stir-fried vegetables and two servings of fresh fruits. Plasma carotenoid levels in subjects significantly increased from 19%-32% for beta-carotene, 15%-47% for lycopene and 59%-88% for beta-crytoxanthin (p < 0.05) from week 1 to 4. However, these concentrations of beta-carotene and lycopene significantly decreased after stopping the consumption of the test meals. This study reveals the importance of a continuous consumption of carotenoid rich foods in order to maintain high levels of plasma carotenoids for the potential prevention of chronic diseases in individuals.

A rice bran oil diet increases LDL-receptor and HMG-CoA reductase mRNA expressions and insulin sensitivity in rats with streptozotocin/nicotinamide-induced type 2 diabetes.

A rice bran oil (RBO) diet can reduce plasma lipids; this was attributed to the specific components, gamma-oryzanol and gamma-tocotrienol, which individually were shown to be hypocholesterolemic; however, the mechanism of their effects on diabetic hyperlipidemia and the development of diabetes is not known. Rats with streptozotocin/nicotinamide-induced type 2 diabetes were divided into control, RO10, and RO15 groups, and fed cholesterol-free diets containing 0, 10, and 15 g RBO with 0, 352, and 528 g gamma-oryzanol and 0, 6.0 and 9.0 mg gamma-tocotrienol/100 g diet for 4 wk. Diabetic rats fed the RBO diet had greater insulin sensitivity (P = 0.02) than rats fed the control diet. Diabetic rats fed the RBO diet also had lower plasma triglyceride (P = 0.003), LDL cholesterol (P = 0.028), and hepatic triglyceride concentrations (P = 0.04), as well as greater fecal neutral sterol and bile acid excretion than those fed the control diet. After 4 wk, there was an approximately 100% (P < 0.001) increase in the abundance of hepatic cholesterol 7alpha-hydroxylase, an 89% (P < 0.001) increase in the hepatic LDL-receptor, and a 50% (P < 0.001) increase in hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA in rats fed the RBO diet compared with those fed the control diet. These findings support the conclusion that a rice bran oil-containing diet can significantly suppress hyperlipidemic and hyperinsulinemic responses in diabetic rats. The high contents of gamma-oryzanol and gamma-tocotrienol in RBO can lead to increased fecal neutral sterol and bile acid excretion, via upregulation of cholesterol synthesis and catabolism.

Chromium yeast supplementation improves fasting plasma glucose and LDL-cholesterol in streptozotocin-induced diabetic rats.

Chromium yeast supplementation has been studied for its ability to improve carbohydrate and lipid abnormalities. There have been some earlier literature-reported studies involving chromium supplementation amongst patients suffering diabetes, but the results would appear to be somewhat varied. Forty male Wistar rats (ten weeks old, 300 g in average body mass) were divided into one of four groups, namely (i) controls; (ii) controls treated with chromium yeast; (iii) diabetic controls; and (iv) diabetic rats treated with chromium yeast. In the present investigation, the effect of a four-week oral administration of chromium yeast (600 microg of Cr/kg body mass/day, by gavage) upon the glucose and lipid metabolism in streptozotocin (STZ)-induced diabetic rats was assessed. Supplemental Cr yeast decreased the fasting blood glucose amongst the STZ-diabetic rats. No significant difference was observed in plasma fructosamine levels of rats treated with chromium yeast compared to control rats. Supplemental Cr yeast did decrease the plasma low-density lipoprotein (LDL)-cholesterol level for the STZ-diabetic rats as compared to controls. We noted no significant effect of chromium supplementation upon plasma high-density lipoprotein (HDL)-cholesterol or triglycerides compared to controls. Treatment with chromium yeast significantly increased the blood and urine chromium levels for both the diabetic and normal rats compared to respective control groups. The results of these studies suggest that Cr yeast decreased the fasting blood glucose and LDL-cholesterol levels in STZ-induced diabetic rats. This raises the possibility that Cr yeast supplementation can be considered to improve carbohydrate and lipid metabolism amongst human patients featuring type 2 diabetes mellitus.

Alleviation of oxidative damage in multiple tissues in rats with streptozotocin-induced diabetes by rice bran oil supplementation.

The possibility of 8-hydroxy-2'-deoxyguanosine (8-OHdG) serving as a sensitive biomarker of oxidative DNA damage and oxidative stress was investigated. Reactive oxygen species (ROS) have been reported to be a cause of diabetes induced by chemicals such as streptozotocin (STZ) in experimental animals. In this study, we examined oxidative DNA damage in multiple tissues in rats with STZ-induced diabetes by measuring the levels of 8-OHdG in the liver, kidney, pancreas, brain, and heart. Levels of 8-OHdG in mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) were also determined in multiple tissues of rats treated with rice bran oil. Levels were 0.19 +/- 0.07, 0.88 +/- 0.30, 1.97 +/- 0.05, and 9.79 +/- 3.09 (1/10(5) dG) in the liver of nDNA of normal rats, nDNA of STZ-induced diabetic rats, mtDNA of normal rats, and mtDNA of STZ-induced diabetic rats, respectively. Levels of mtDNA of 8-OHdG were 10 times higher than those of nDNA in multiple tissues. Significant reductions in mtDNA 8-OHdG levels were seen in the liver, kidney, and pancreas of diabetic rats treated with rice bran oil compared with diabetic rats without intervention. Our study demonstrated that oxidative mtDNA damage may occur in multiple tissues of STZ-induced diabetics rats. Intervention with rice bran oil treatment may reverse the increase in the frequency of 8-OHdG.

Antioxidant effects of chromium supplementation with type 2 diabetes mellitus and euglycemic subjects.

To determine the effects of chromium (Cr) supplementations on oxidative stress of type 2 diabetes and euglycemic (EU) subjects, adult having HbA(1C) values of <6.0% (EU), 6.8-8.5% (mildly hyperglycemic, MH), and >8.5% (severely hyperglycemic, SH) were supplemented for 6 months with 1000 microg/day of Cr (as Cr yeast) or with a placebo. In the beginning, the levels of the plasma Cr in the MH and SH groups were 25-30% lower than those of the EU subjects. The values of thiobarbituric acid reactive substances (TBARS) and total antioxidative status (TAS) of the MH and SH groups were significantly higher than those of the EU ones. Following supplementations, the levels of plasma TBARS in the Cr groups of MH and SH groups were significantly decreased (the inverse was found in the EU) and showed no significant changes in the placebo group. The levels of plasma TAS in the Cr groups of EU and MH were significantly decreased (the inverse was found in the SH) and showed no significant changes in the placebo group. No significant difference was found in the antioxidant enzyme (superoxide dismutase, glutathione peroxidase, catalase) activities during supplementations. These data suggest that Cr supplementation was an effective treatment strategy to minimize increased oxidative stress in type 2 diabetes mellitus patients whose HbA(1C) level was >8.5%, and the Cr in EU groups might act as a prooxidant.

Interactions of lipid metabolism and intestinal physiology with Tremella fuciformis Berk edible mushroom in rats fed a high-cholesterol diet with or without Nebacitin.

Male adult Wistar rats were randomly divided into six groups in a 2 x 3 factorial design and fed diets containing different levels of Tremella fuciformis Berk (TFB) dietary fiber (0, 5, or 10%) and 1 g of cholesterol/100 g of diet with or without 0.7% Nebacitin for 4 weeks. TFB contained 6.2% soluble dietary fiber and 57.3% insoluble dietary fiber. The results showed that the serum LDL-cholesterol, hepatic total cholesterol, and triglyceride levels were significantly decreased (P < 0.05) in the rats fed diets with TFB content with or without Nebacitin. However, the serum total cholesterol, VLDL-cholesterol, and triglyceride levels were significantly decreased (P < 0.05) by Nebacitin. In feces, the presence of TFB (T5, T10, AT5, and AT10) in the diet significantly increased the total neutral steroids and bile acid excretions and undigested fiber concentrations as compared to T0 or AT0. In the small intestine, the Nebacitin diets increased the weights of both cecum and colon-rectum contents and lowered short-chain fatty acid (SCFA) concentrations of serum and cecal contents more than no Nebacitin diets did. It was suggested that the hypocholesterolemic effect of TFB dietary fiber may be mediated by the increase in fecal neutral steroids and total bile acids excretion and the increase in SCFA productions. The TFB edible mushroom dietary supplement altered the intestinal physiology of the rats.