Resistant starch fraction prepared from kintoki bean affects gene expression of genes associated with cholesterol metabolism in rats

Feeding rats beans with resistant starch reduces the serum cholesterol concentration; however, the mechanism is not fully understood. We examined the effects of resistant starch of kintoki (Phaseolus vulgaris, variety) bean on serum cholesterol and hepatic mRNAs in rats. Male F344/Du Crj rats were fed a cholesterol-free diet either with 5 g of cellulose powder (control)/100 g or 5 g of pancreatin-resistant fraction prepared from kintoki bean (kintoki)/100 g diet for 4 weeks. There were no differences in the body weight gain, food intake, liver weight, and mass of cecum contents between the groups. Serum total cholesterol, very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) + low density lipoprotein (LDL)-cholesterol, and high density lipoprotein (HDL)-cholesterol levels in the kintoki group were significantly (at least P < 0.05) lower than in the control group throughout the feeding period. There was no difference in the serum triglyceride concentration between two groups throughout the feeding period. Total hepatic cholesterol in the control group was significantly (P < 0.01) lower than in the kintoki groups. Fecal bile acid, cecal acetate, propionate and n-butyrate concentrations in the kintoki group all were significantly (P < 0.05) higher than in the control group. Likewise, hepatic cholesterol 7alpha-hydroxylase, LDL receptor, and SR-B1 mRNA levels in the kintoki group were significantly (P < 0.05) higher than in the control group. The results suggest that resistant starch of kintoki bean reduces serum cholesterol level by increasing hepatic LDL receptor, SR-B1, and cholesterol 7alpha-hydroxylase mRNAs.     

A mixture of organisms affects cholesterol metabolism together with rat cecal flora.

The effects of a mixture of organisms on cecal fermentation and cholesterol metabolism in sham-operated and cecectomized rats were investigated. Male F344 rats, allocated into four groups: cecectomized rats fed a mixture of organisms (CEMO), cecectomized rats fed rice bran (CERB), sham-operated rats fed a mixture of organisms (SHMO), and sham-operated rats fed rice bran (SHRB) for 4 weeks. The diets had 0.5% cholesterol and 0.125% sodium cholate added. There were no significant differences in the body weight gain and food intake among the groups. The cecal pH in the SHMO group was significantly lower than that in the other groups. The total cholesterol and (VLDL + IDL + LDL)-cholesterol concentrations in serum were significantly lower in the SHMO group than that in the SHRB group, and the triacylglycerol concentration in the sham-operated rats tended to decrease compared to the cecectomized rats. The fecal cholesterol excretion in the CERB group was higher than that in the other groups, and that in the SHMO group was significantly higher than in the SHRB group. The acetic acid, propionic acid, n-butyric acid, and total short-chain fatty acid concentrations in the cecum contents were significantly higher in the SHMO group than those in the other groups. Streptococcus, Bifidobacterium, and Lactobacillus in the SHMO group tended to be higher than the other groups and Bacteroidaceae in the CEMO and CERB groups were significantly higher than that in the SHMO group. The results demonstrate that the mixture of organisms was fermented with the cecal contents and that the metabolites such as short-chain fatty acid lowered the serum total cholesterol and liver cholesterol concentrations in the rats fed a cholesterol-containing diet.     

Hypocholesterolemic effect of protein prepared from Phaseolus aconitifolius (Jacq.)

Hypocholesterolemic effect of protein concentrate (PC) prepared from moth bean (Phaseolus aconitifolius Jacq.) seeds relative to that of pigeon pea PC and casein were investigated in rats fed on hypercholesterolemic diet containing two per cent cholesterol. The test diets containing casein and legume PCs at 10% protein level were fed to albino rats (Wistar strain) for 45 days. Compared to casein, the two legume PCs produced significantly lower levels of liver total lipid and cholesterol levels except that pigeon pea PC produced only non-significant decrease of total cholesterols in the heart. In addition, the legume PCs produced significantly lower levels of serum triglycerides and total lipids as well as lower levels of total and low-density lipoprotein cholesterol. Moreover, only moth bean PC produced a significantly higher level of serum high-density lipoprotein cholesterol. Generally, PC of moth bean seemed to be more potent at lowering the elevated hepatic and serum lipids and cholesterol levels, which were attributed to the amino acid profile of this lesser-known legume as these indices well correlated with serum cholesterol levels.     

Comparative hypocholesterolemic effects of five animal oils in cholesterol-fed rats

The hypocholesterolemic efficacy of various animal oils was compared in rats given a cholesterol-enriched diet. After acclimatization for one week, male F344 DuCrj rats (8 weeks of age) that had been fed with a conventional diet were assigned to diets containing 5% of oil from emu (Dromaius), Japanese Sika deer (Cervus nippon yesoensis, Heude), sardine, beef tallow, or lard with 0.5% cholesterol for 6 weeks. After this feeding period, the concentrations of serum total cholesterol and of very-low-density lipoprotein + intermediate-density lipoprotein + low-density lipoprotein-cholesterol in the sardine oil group were significantly lower than those in the other groups. The serum high-density lipoprotein-cholesterol concentration in the Japanese Sika deer oil group was significantly higher than that in the other groups. The atherosclerotic index and liver cholesterol concentration in the sardine oil and Japanese Sika deer oil groups were significantly lower than those in the other groups. The fecal cholesterol excretion by the Japanese Sika deer oil group was significantly higher than that of the other groups, except for the sardine oil group, and the fecal bile acid excretion by the sardine oil group was significantly higher than that of the other groups, except for the lard group. These results suggest that Japanese Sika deer oil reduced the atherosclerotic index and liver cholesterol concentration in the presence of excess cholesterol in the diet as well as sardine oil did by increasing the excretion of cholesterol from the intestines of rats.     

The effect of diabetes mellitus on aortic prostanoid synthesis and serum cholesterol levels in the rat fed a high cholesterol diet

The effect of diabetes mellitus on serum cholesterol and aortic microsomal prostanoid synthesis was studied in cholesterol fed male Lewis rats. Normal, diabetic and diabetic rats treated with pancreatic islets were divided into three diet subgroups, control diet, control +2% cholesterol for 8 weeks and control +2% cholesterol diet for 16 weeks. Serum glucose levels were elevated three-fold in the diabetic group compared to normal. Treatment with islets restored serum glucose to normal levels in diabetic rats. The 2% cholesterol diet did not significantly alter serum glucose levels in any of the groups. Body weights in the diabetic group were significantly lower than normal or diabetic rats treated with islets. Feeding 2% cholesterol for 16 weeks significantly increased weight in normal and islet treated diabetic rats but not in the diabetic group. Aortic microsomal prostanoid synthesis was similar in all experimental groups with 6-keto-PGF1 alpha (PGI2 metabolite) being the major product synthesized in all groups. Aortic microsomal prostanoid levels were not altered by the 2% cholesterol diet. Serum cholesterol levels increased 14-fold in the diabetic group which returned to the normal level in the diabetic animals treated with islets. These data show that diabetes does not alter aortic microsomal prostanoid levels in the rat. However, diabetes significantly increased serum cholesterol levels which were reversed by islet transplantation.     

参麦注射液对高脂血症大鼠血脂的调节作用

目的：通过观察参麦注射液对高脂血症模型大鼠的血脂水平和一系列相关生化指标的影响,探讨其对高脂血症模型大鼠血脂的调节作用。方法：30只SPF级雄性SD大鼠用基础饲料适应性喂养1周,随机分为3组（n=10）：对照组,模型对照组,参麦注射液组。对照组用基础饲料喂养,模型对照组和参麦注射液组用高脂饲料喂养,每周测定动物体重一次。参麦注射液组每天给予2次参麦注射液10 ml/kg,连续灌胃45 d。之后测定大鼠血清总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白（HDL-C）、低密度脂蛋白（LDL-C）、超氧化物歧化酶（SOD）、谷胱甘肽过氧物酶（GSH-Px）、丙二醛（MDA）、脂蛋白酯酶（LPL）和肝酯酶（HL）活性。结果：模型对照组大鼠体重、血清中TC、TG、LDL-C和MDA水平较对照组明显升高（P<0.01）,而HDL-C、SOD、GSH-Px、LPL和HL水平明显降低（P<0.01）。参麦注射液组大鼠体重、血清中TC、TG、LDL-C和MDA水平较模型对照组明显降低（P<0.01）,而HDL-C、SOD、GSH-Px、LPL和HL水平明显升高（P<0.05,P<0.01）。结论：参麦注射液对高脂模型大鼠的血脂具有较明显的调节作用,并具有抗脂质过氧化的作用。     

Effect of an adzuki bean extract on hepatic anti-oxidant enzyme mRNAs in D-galactosamine-treated rats

In acute hepatic injury tests, an adzuki bean extract decreased D-galactosamine (GalN)-induced alterations in the serum alanine aminotransferase and aspartate aminotranferase activities to about 37% and 25%, respectively, although there were no significant differences in these activities between the GalN-treated group with the adzuki bean extract and the GalN-treated group without the adzuki bean extract. Furthermore, the hepatic glutathione peroxidase, glutathione reductase, and Mn- and Cu,Zn-superoxide dismutase mRNA levels in the GalN-treated group with the adzuki bean extract were higher than those in the control group and GalN-treated group without the adzuki bean extract.     

Potato powders prepared by successive cooking-process depending on resistant starch content affect the intestinal fermentation in rats

The effects of resistant starch (RS) in dry potato powders prepared by various processes on intestinal fermentation in rats were assessed. Rats were fed raw potato powder (RP), blanched potato powder (BP), steamed potato powder (SP), or drum-dried potato powder (DP) for 4 weeks. The cecal RS content was significantly higher in the RP group than in the control diet (CN) group and other dry potato powder groups. Cecum pH was significantly lower in the RP group compared to the CN group, and was also significantly lower than that in the SP, BP, and DP groups. Lactic acid bacteria levels in the RP group were significantly higher than those in the CN group, and levels in the SP group also increased relative to the control group. Lactobacillus levels in the RP group were higher than in the CN and other dry potato powder groups. Cecal short-chain fatty acid (SCFA) concentrations in the RP group followed by the SP group exhibited significantly higher levels relative to the control levels. Dry potato powders containing RS produced during the cooking process may represent a useful food material that increases intestinal concentrations of SCFA and enhances the growth of certain lactic acid bacteria.     

Safety and efficacy of adzuki bean extract in subjects with moderate to high LDL-C: a randomized trial

The safety and efficacy of polyphenol-containing adzuki bean extract on lipid metabolism were evaluated in human subjects in an 8-week, randomized, double-blind, placebo-controlled, parallel intervention study. No adverse effects were observed in the participants receiving adzuki bean extract. The adzuki bean group showed a significant increase in the ΔHDL-C concentration compared with the placebo group after 4 weeks of intervention (3.76 ± 7.79 mg/dL vs. ?0.08 ± 6.03 mg/dL), respectively, and both groups showed reduced ?HDL-C concentrations, with the adzuki bean extract group showing a return to the baseline levels (0.36 ± 5.36 mg/dL) and the placebo group showing a decrease to below the baseline levels (?3.17 ± 7.79 mg/dL) at week 8. This short-term study represents the first step in establishing the practicality, safety, and plausibility of HDL-C maintaining effects of adzuki bean extract in human subjects.     

Effect of red mold rice supplements on serum and meat cholesterol levels of broilers chicken

Monacolin K is a secondary metabolite produced by Monascus species. It was found that it is able to decrease cholesterol levels. In this study, red mold rice (RMR) was added to the diet of Arbor Acres broiler chickens, and the cholesterol level in meat, as well as the concentration of triglyceride, the high-density lipoprotein cholesterol (HDL-C), and the low-density lipoprotein cholesterol (LDL-C) in the serum were evaluated. Four-week-old broilers are studied and divided into four groups in that each group contains 15 subjects. A 3-week experimental feeding trial was conducted in which three groups of broilers were fed 2.0, 5.0, and 8.0% of RMR (RMR groups) within their diet, respectively, and the result was compared to the control group. The results indicated that for each RMR group, the cholesterol content was significantly lower than that of the control group; in addition, their meat products contain higher level of unsaturated fatty acids. Triglyceride and cholesterol concentration in serum was also found to be considerably lower in RMR groups when compared to control group. Finally, in RMR groups, HDL-C/LDL-C and HDL-C/cholesterol ratios were all higher than those of the control group. In short, the results demonstrated that the cholesterol levels could be lowered by adding RMR to the diet of chickens.     

Effect of Soymilk Fermented with Lactobacillus plantarum P-8 on Lipid Metabolism and Fecal Microbiota in Experimental Hyperlipidemic Rats

We recently identified a novel probiotic strain Lactobacillus plantarum P-8 (L. plantarum P-8), which has been characterized in detail with regard to its probiotic potential. In the present study, soymilk fermented with L. plantarum P-8 was examined for its effects on diet-induced hyperlipidemia in Wistar rats. The experimental animals were divided into four groups: control group (C group), model group (M group), soymilk group (SM group) and fermented soymilk group (FSM group). The serum lipid levels, hepatic fat deposition, serum oxidative stress parameters, hepatic marker enzymes levels, organ indices, gut bacteria and fecal fat contents were analyzed. Fermented soymilk reduced the concentration of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum, with a significant elevation in high-density lipoprotein cholesterol (HDL) concentration. Our results also suggested the beneficial effects of fermented soymilk on the liver function, hyperlipidemia-induced oxidative stress and intestinal bacteria. Moreover, fermented soymilk could enhance the fecal excretion of TC, TG and bile acids. These findings demonstrated that soymilk fermented with L. plantarum P-8 was effective in improving the lipid metabolism in hyperlipidemic rats. The hypolipidemic effect of fermented soymilk was partly due to the inhibition of dietary fats absorption and regulation of fecal fats excretion mediated by gut bacteria.     

Relationship between exercise-induced changes in serum and hepatic cholesterol metabolism in rats

The purpose of the present study was to determine the effects of exercise training on serum cholesterol level and hepatic cholesterol metabolism in rats. Twenty-four male Wistar strain rats, aged 6 weeks, were assigned to one of three experimental groups; control (n = 8), exercised 20 minutes a day (E20, n = 8), and exercised 60 minutes a day (E60, n = 8). Rats were sacrificed after ten weeks exercise. The levels of serum total cholesterol and HDL-cholesterol in the group E20 and group E60 were lower than that of control, 8% and 24% (p less than 0.01) for the total cholesterol and 10% and 24% (p less than 0.05) for the HDL-cholesterol, respectively. The activity of HMG-CoA reductase in liver microsome was significantly higher in the group E60 than that of control. The hepatic microsomal HMG-CoA reductase activity was negatively related to serum total (r = -0.62, p less than 0.01) and HDL (r = -0.58, p less than 0.01) cholesterol levels. From these results, we concluded that the enhancement of the cholesterol metabolism in the liver by exercise is a major course of the exercise-induced change in serum cholesterol of Wistar rats.     

不同脂肪高脂日粮诱发胰岛素抵抗综合征大鼠模型的比较

目的分析猪油、豆油、氢化椰子油、乳脂四种不同脂肪的高脂日粮分别诱发胰岛素抵抗综合征（IRS）大鼠的血液生化指标差异,为此类模型的建立及实验研究提供参考。方法雄性SD大鼠随机分为5组,对照组给予普通日粮,高脂组给予脂肪热量比相同的高脂日粮。喂养6周,每两周测定空腹血糖、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-c）、总胆固醇（TC）、胰岛素,根据胰岛素敏感性指数（ISI）=ln1/（FPG×FINS）评定大鼠的胰岛素敏感性。结果6周后,猪油组、乳脂组、豆油组血清胰岛素均显著高于对照组（P〈0.05）;乳脂组血清TG显著高于其它高脂组（P〈0.05）;高脂组血清HDL-c均显著低于对照组（P〈0.05）并以豆油组下降幅度最大;猪油组、乳脂组ISI显著低于对照组（P〈0.05）;而各组间血清总胆固醇、血糖及体重无明显差异（P〉0.05）。结论4种高脂日粮诱发IRS大鼠模型的综合效果依次为乳脂、猪油、豆油、氢化椰子油。     

Suppression of Plasma Cholesterol Elevation by Okara Tempe in Rats

Okara Tempe (OT), an Indonesian fermented traditional food, which is the fermented okara (insoluble residues of homogenized soybean, OC) by Rhizopus oligosporus, and which is interested in as a new high fiber and low energy soybean foodstuff. In this study, the effects of protein and dietary fiber (DF) of OT on sterol metabolism in rats were investigated. When rats were fed by OT or casein as protein source, the cholesterol and bile acid levels in plasma was significantly lower in OT fed group than in casein fed group. When rats were fed by OT or OC as DF source, in OT fed group, the cholesterol and bile acid levels in plasma was similar to them in OC fed group, but the cholesterol level in liver was lower than it in OC fed group, and the excretion of cholesterol and bile acid in feces was larger than in OC fed group. These results suggest that the cholesterol lowering action of OT in plasma would arise from the complementary action of protein and water soluble DF in OT.     

Effect of pectin and amidated pectin on cholesterol homeostasis and cecal metabolism in rats fed a high-cholesterol diet

Two experiments were performed to compare the effect of pectin and its hydrophobic derivatives on homeostasis of cholesterol and cecal metabolism in male young rats. Control rats were fed a diet supplemented with palm fat and cholesterol (50 and 10 g/kg, respectively). Rats of other groups were fed the same diet containing citrus pectin or octadecylpectinamide (60 g/kg). Diets were fed for 4 weeks. In experiment I, pectinamide of lower degree of amidation (30 %) increased serum HDL cholesterol from 1.20 to 1.43 micromol/ml (p>0.05) at the expense of other cholesterol fractions. In experiment II, pectinamide of a higher degree of amidation (53 %) significantly decreased total serum cholesterol from 2.08 to 1.67 micromol/ml. Amidated pectins at both levels of substitution significantly decreased hepatic concentrations of cholesterol and fat. In both experiments the relative weight of cecum in the pectinamide group was significantly lower than in pectin group. The highest cecal concentrations of short-chain fatty acids (SCFA) were found in rats fed a diet with pectin (133.2 and 129.3 micromol/g in experiment I and II, respectively). In other groups, cecal SCFA was significantly (pectinamide groups) or non-significantly (controls) lower. In wet feces, SCFA concentrations were higher and butyrate molar proportions lower than in corresponding cecal contents. Pectinamide of a lower or higher degree of substitution significantly increased fecal content of cholesterol from 18.5 and 17.3 micromol/g in controls to 31.8 and 28.0 micromol/g, respectively. Corresponding concentrations of coprostanol were decreased. Effects of pectin on cholesterol homeostasis were absent or marginal. Histological examination revealed that hepatic tissue of control and pectin-fed rats was infiltrated with lipids. The Sudan black-positive material was absent in the liver of rats fed pectinamides. No pathological changes of liver tissue were apparent. In summary, hydrophobic amidated pectins significantly altered cholesterol homeostasis in rats and might be considered as a clinically effective hypocholesterolemic agent. Low cecal SCFA concentrations in rats fed pectinamides suggest that amidation of pectin had decreased its fermentability.     

Potato and soy peptide diets modulate lipid metabolism in rats

Dietary plant and animal peptides have been shown to reduce serum lipids. However, the potential of food-derived peptides has yet to be fully elucidated. We investigated the physiological importance of potato peptides in rats fed on a cholesterol-free diet containing 20% potato peptides (PP), when compared with two diets containing either 20% casein (CN) or 20% soy peptides (SP). The high-density lipoprotein (HDL)-cholesterol (+13.8%) and serum triglyceride (-38%) concentrations in the PP-fed group, non-HDL-cholesterol level in the PP- (-22.5%) and SP- (-15.7%) fed groups, and serum total cholesterol concentration (-12%) in the SP-fed group, were significantly different from the control group at the end of the experiment. The fecal excretion of neutral and acidic sterols was higher in the PP- and SP-fed groups, respectively, relative to the control group. These results indicate that the observed changes in the serum cholesterol levels in rats fed on soy and potato peptide appear to have been due to different mechanisms.     

Effect of γ-Aminobutyric Acid-enriched Tempeh-like Fermented Soybean (GABA-Tempeh) on the Blood Pressure of Spontaneously Hypertensive Rats

GABA-enriched tempeh-like fermented soybean (GABA-tempeh) was supplemented to the AIN-76 diet and fed for 2 months to spontaneously hypertensive rats (SHRs), an animal model of spontaneously developed hypertension, to compare the antihypertensive activity with that of authentic GABA. The elevation of systolic blood pressure in SHRs was significantly retarded in the GABA-tempeh group as well as that with authentic GABA when compared with the controls, and the effect lasted for two months of the feeding period. The blood urea nitrogen level tended to be higher in the control group than in the GABA-supplemented groups. On the other hand, no effect was apparent on the plasma levels of cholesterol, triacylglycerol and glucose, or on the urinary excretion of Na and K.     

Reduction by phytate-reduced soybean beta-conglycinin of plasma triglyceride level of young and adult rats.

This study examined the effects of soybean beta-conglycinin, from which phytate was mostly removed, on the plasma lipids in young and adult rats. Male Wistar young (6 week-old) and adult (21 week-old) rats were fed high cholesterol diets containing 20% casein, soy protein isolate (SPI), or soybean beta-conglycinin for 10 days. In young rats, although the food intake of the beta-conglycinin group was higher than those of the casein and SPI groups, the weight gain was significantly lower than those of the other groups. However, in adult rats, the weight gain was not different among the groups. In young and adult rats, relative liver weights of SPI and beta-conglycinin groups were significantly lower than that of the casein group, and the degree of the reduction was more marked in the beta-conglycinin group than in the SPI group. In young rats, the plasma triglyceride level was significantly lower in the SPI and beta-conglycinin groups than that in the casein group. In addition, the plasma triglyceride level of the beta-conglycinin group was significantly lower than that of the SPI group. Plasma total cholesterol levels of the SPI and beta-conglycinin groups were significantly lower than that of the casein group. However, there was little difference in the lowering effect between SPI and beta-conglycinin. These results indicate that soybean beta-conglycinin may have lowering functions not only on plasma total cholesterol level, but also on plasma triglyceride level.     

Cholesterol-lowering effects of maitake (Grifola frondosa) fiber, shiitake (Lentinus edodes) fiber, and enokitake (Flammulina velutipes) fiber in rats

The effects of mushroom fibers on serum cholesterol and hepatic low-density lipoprotein (LDL) receptor mRNA in rats were investigated. Rats were fed a cholesterol-free diet with 50 g/kg cellulose powder (CP), 50 g/kg maitake (Grifola frondosa) fiber (MAF), 50 g/kg shiitake (Lentinus edodes) fiber (SF), or 50 g/kg enokitake (Flammulina velutipes) fiber (EF) for 4 weeks. There were no significant differences in the body weight, food intake, liver weight, cecum weight, and cecum pH among the groups. Cecal acetic acid, butyric acid, and total short-chain fatty acid (SCFA) concentrations in the SF and EF groups were significantly higher than those in the other groups. The serum total cholesterol concentration in the CP group was significantly higher than that in the MAF and EF groups. The very LDL (VLDL) + intermediate-density lipoprotein (IDL) + LDL-cholesterol concentration in the CP group was significantly higher than that in the MAF, SF, and EF groups, whereas the high-density lipoprotein (HDL)-cholesterol concentration in the EF group was significantly lower than that in the other groups at the end of the 4-week feeding period. The hepatic LDL receptor mRNA level in the EF group was significantly higher than that in the CP group. The fecal cholesterol excretion in the MAF, SF, and EF groups was significantly higher than that in the CP group. The results of this study demonstrate that MAF and EF lowered the serum total cholesterol level by enhancement of fecal cholesterol excretion, and in particular, by enhancement of hepatic LDL receptor mRNA in EF group.     

Anti-hyperlipidemic effect of soybean extract fermented by Bacillus subtilis MORI in db/db mice

The purpose of this study was to investigate the anti-hyperlipidemic effect of soy bean extract solution fermented by Bacillus subtilis MORI (BTD-1E) in obese db/db mice. Eight-week-old male db/db mice were administered 33.3 mg/kg BTD-1E solution orally once a day for four weeks. The BTD-1E group showed significantly lower body weight compared with the db control group (P<0.05). The BTD-1E group showed significantly lower serum total cholesterol and LDL cholesterol levels compared with the db control group, respectively (P<0.05, P<0.01). The BTD-1E group showed significantly decreased liver weight relative to final body weight compared with the db control group (P<0.01). After four weeks of BTD-1E administration, lipid droplets in the liver were apparently decreased in the BTD-1E group compared to the db control group. In summary, our results suggest that BTD-1E has an anti-hyperlipidemic effect in the obese mouse model.