Hormone-sensitive Lipase Levels Research Articles

Comparative evaluation on the effects of dietary docosahexaenoic acid on growth performance, fatty acid profile and lipid metabolism in two sizes of abalone Haliotis discus hannai Ino

A 93-day feeding trial was conducted to comparatively evaluate the effects of graded levels of dietary docosahexaenoic acid (DHA) (0.06%, 0.46%, 0.99%, 1.57%, 1.95% and 2.25%) on growth performance, fatty acid profile and lipid metabolism in two different sizes of abalone Haliotis discus hannai (initial body weight: 2.83 ± 0.03 g and 26.81 ± 0.17 g, respectively). Results showed that 0.99%–1.95% of dietary DHA contents significantly increased the weight gain rate (WGR) in large size of abalone (P < 0.05), while the WGR in small size of abalone was significantly increased by the 0.46%–0.99% of dietary DHA contents (P < 0.05). There was no significant difference in the survival of abalone among all the six groups (P > 0.05). Based on the WGR, the optimal dietary DHA content for large and small size abalone were estimated to be 1.32% and 0.88%, respectively. Dietary DHA supplementation significantly increased the activities of trypsin and lipase in the intestine, the content of ∑n-3 polyunsaturated fatty acids (PUFA) and ∑n-3/n-6 PUFA ratio in digestive gland in both sizes of abalone (P < 0.05). The total glyceride concentration in the cell-free hemolymph in two sizes of abalone was decreased. In large size of abalone, 1.95%–2.25% of dietary DHA reduced the lipid content in digestive gland. Lipogenic genes in the digestive gland, including sterol regulatory element binding protein-1 (srebp-1) and diacylglycerol acyltransferase, were up-regulated in the 0.46%–1.57% DHA group (P > 0.05). The mRNA expression levels of hormone sensitive lipase (hsl) and carnitine palmitoyl transferase-1 (cpt-1) of lipolytic genes in the digestive gland were significantly increased in the 0.46% and 0.99% DHA groups, respectively (P < 0.05). In the small size of abalone, the lipid content in digestive gland and soft body decreased significantly in the 1.95% and 2.25% DHA groups, respectively (P < 0.05). Lipogenic genes in the digestive gland, including srebp-1 and acetyl-CoA carboxylase, were significantly down-regulated in the 0.99%–1.95% DHA group (P < 0.05). Lipolytic genes in the digestive gland, including hsl and cpt-1, were not significantly different from the control group (P > 0.05). In conclusion, based on the WGR, the dietary DHA requirements for large and small size of abalone were estimated to be 1.32% and 0.88%, respectively. Dietary DHA reduced lipid deposition in both sizes of abalone by promoting lipid decomposition and inhibiting lipid synthesis.

Cloning and Molecular Characterization of HSL and Its Expression Pattern in HPG Axis and Testis during Different Stages in Bactrian Camel.

Hormone-sensitive lipase (HSL) is a key enzyme in animal fat metabolism and is involved in the rate-limiting step of catalyzing the decomposition of fat and cholesterol. It also plays an important regulatory role in maintaining seminiferous epithelial structure, androgen synthesis and primordial germ cell differentiation. We previously reported that HSL is involved the synthesis of steroids in Bactrian camels, although it is unclear what role it plays in testicular development. The present study was conducted to characterize the biological function and expression pattern of the HSL gene in the hypothalamic pituitary gonadal (HPG) axis and the development of testis in Bactrian camels. We analyzed cloning of the cDNA sequence of the HSL gene of Bactrian camels by RT-PCR, as well as the structural features of HSL proteins, using bioinformatics software, such as ProtParam, TMHMM, Signal P 4.1, SOPMA and MEGA 7.0. We used qRT-PCR, Western blotting and immunofluorescence staining to clarify the expression pattern of HSL in the HPG axis and testis of two-week-old (2W), two-year-old (2Y), four-year-old (4Y) and six-year-old (6Y) Bactrian camels. According to sequence analysis, the coding sequence (CDS) region of the HSL gene is 648 bp in length and encodes 204 amino acids. According to bioinformatics analysis, the nucleotide and amino acid sequence of Bactrian camel HSL are most similar to those of Camelus pacos and Camelus dromedarius, with the lowest sequence similarity with Mus musculus. In adult Bactrian camel HPG axis tissues, both HSL mRNA and protein expression were significantly higher in the testis than in other tissues (hypothalamus, pituitary and pineal tissues) (p < 0.05). The expression of mRNA in the testis increased with age and was the highest in six-year-old testis (p < 0.01). The protein expression levels of HSL in 2Y and 6Y testis were clearly higher than in 2W and 4Y testis tissues (p < 0.01). Immunofluorescence results indicate that the HSL protein was mainly localized in the germ cells, Sertoli cells and Leydig cells from Bactrian camel testis, and strong positive signals were detected in epididymal epithelial cells, basal cells, spermatocytes and smooth muscle cells, with partially expression in hypothalamic glial cells, pituitary suspensory cells and pineal cells. According to the results of gene ontology (GO) analysis enrichment, HSL indirectly regulates the anabolism of steroid hormones through interactions with various targets. Therefore, we conclude that the HSL gene may be associated with the development and reproduction of Bactrian camels in different stages of maturity, and these results will contribute to further understanding of the regulatory mechanisms of HSL in Bactrian camel reproduction.

The involvement of hormone-sensitive lipase in all-trans retinoic acid induced cleft palate.

Abnormally high concentrations of all-trans retinoic acid (atRA) induce cleft palate, which is accompanied by abnormal migration and proliferation of mouse embryonic palatal mesenchyme (MEPM) cells. Hormone-sensitive lipase (HSL) is involved in many embryonic development processes. The current study was designed to elucidate the mechanism of HSL in cleft palate induced by atRA. To establish a cleft palate model in Kunming mice, pregnant mice were administered atRA (70 mg/kg) by gavage at embryonic Day 10.5 (E10.5). Embryonic palates were obtained through the dissection of pregnant mice at E15.5. Hematoxylin and eosin (H&E) staining was used to evaluate growth changes in the palatal shelves. The levels of HSL in MEPM cells were detected by immunohistochemistry, quantitative real-time reverse transcription-polymerase chain reaction (qRT‒PCR) and western blotting. RNAi was applied to construct vectors expressing HSL small interference RNAs (siRNAs). The vectors were transfected into MEPM cells. Cell proliferation and migration were evaluated by the cell counting kit-8 (CCK-8) assay and wound healing assay, respectively. The palatal shelves in the atRA group had separated at E15.5 without fusing. In MEPM cells, the expression of HSL was reversed after atRA treatment, which caused cleft palate in vivo. In the atRA group, the proliferation of HSL siRNA-transfected cells was remarkably promoted, and the migration rate significantly increased in the HSL siRNA-transfected MEPM cells. These results suggested that HSL may be involved in cleft palate induced by atRA and that atRA enhances HSL levels to inhibit embryonic palate growth.

ВЛИЯНИЕ МИОКИНОВ НА СОДЕРЖАНИЕ ГОРМОН-ЧУВСТВИТЕЛЬНОЙ ЛИПАЗЫ В МСК И КЛЕТКАХ АДИПОГЕННОЙ ДИФФЕРЕНЦИРОВКИ

The basic metabolic process associated with white and beige/brown adipose tissues is lipolysis – the sequential enzymatic process of the hydrolysis of triglycerides in the adipose tissue. It has been repeatedly shown that physical activity activates lipolysis. It has recently been shown that skeletal muscles have an endocrine role; producing a host of myogenic hormones – myokines. Current literature has an incomplete understanding of the interdependent relationship between skeletal muscles and adipose tissue. We researched the influence of myocyte secreted cytokines (myokines) – meteorin-like protein (METRNL) and β-aminoisobutyric acid (BAIBA), and the adrenergic agonist isoproterenol on the levels of total and phosphorylated (Ser552) hormone sensitive lipase (HSL) in adipose tissue derived mesenchymal stromal cells (MSCs) and the cellular products of their adipogenic differentiation. The MSCs were obtained from 5 healthy donors. The adipogenic differentiation protocol was carried out for a span of 21 days. After procuring the adipocyte cultures, the following stimulators were added – 5 μM METRNL, 5 μM BAIBA, and 5 μM isoproterenol. With the help of western blot, the change in the amount of total and activated levels of HSL were monitored in cells of three different adipogenic differentiation protocols in MSCs. We observed that HSL and its activated form are produced in cell cultures induced with factors for white, beige, and brown adipogenic differentiation.

Preventive Effects of Different Aerobic Exercise Intensities on the Decline of Cognitive Function in High-Fat Diet-Induced Obese Growing Mice.

Background and Objectives: The purpose of this study was to elucidate the effects of different exercise intensities in preventing the decline of cognitive function and lipolysis associated with a high-fat diet-induced obesity in growing mice. Material and Methods: Forty male C57BL/6 mice, aged 4 weeks, were divided into the normal diet (CO, n = 10) and high-fat diet (HF, n = 30) groups to induce obesity for 8 weeks. Subsequently, the HF group was subdivided equally into the HF, HF + low-intensity training (HFLT), and HF + high-intensity training (HFHT) groups, and mice were subjected to treadmill training for 8 weeks. Results: Following the 8-week training intervention, body weight and fat mass were significantly lower in the training groups than in the HF group (p < 0.05). Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase levels were significantly higher in the training groups than in the HF group (p < 0.05), and the ATGL and HSL levels were significantly higher in the HFHT group than in the HFLT group (p < 0.05). The Y-maze test showed that the training groups had a higher number of total entries and percent alternation than the HF group (p < 0.05). Hippocampal nerve growth factor, brain-derived neurotrophic factor, and neurotrophin-3 levels were significantly higher in the training group than in the HF group (p < 0.05). However, there was no significant difference according to the exercise intensity among the groups. Conclusions: The results of this study suggested that low-intensity exercise is as effective as a high-intensity exercise in preventing the decline of cognitive function and lipolysis, and far more effective in terms of an expected efficiency of workload and prevention of side effects.

Metabolic Effects of Testosterone Hormone Therapy in Normal and Orchiectomized Male Rats: From Indirect Calorimetry to Lipolytic Enzymes.

Background and Aim Changes in total energy expenditure (TEE) and substrate metabolism may help explain the metabolic actions of testosterone (T). This study measured respiratory quotient (RQ), TEE, ghrelin, insulin, and key lipolysis enzyme concentrations in relation to body weight (wt) and food intake (FI) in both normal and bilaterally orchiectomized rats with/without T treatment. Methods In total, thirty-two male Wistar rats (300–400 g) were divided into four groups (n = 8/group), including (a) sham-operated and vehicle-injected group (Sham), (b) T-treated sham group (T-Sham) for which sham-operated rats were injected with IM testosterone undecanoate (100 mg/kg, for one week), (c) orchiectomy and vehicle-injected group (Orch), and (d) T-replaced orchiectomy group (T-Orch). After one week, FI and wt were automatically recorded, indirect calorimetry parameters were measured, and blood samples were collected to measure T, ghrelin, insulin, growth hormone (GH), glucose, hormone-sensitive lipase (HSL), adipocyte triglyceride lipase (ATGL), free fatty acids (FFA), and lipid profiles. Results Orchiectomy decreased ghrelin, GH, and insulin levels, increased TEE and RQ, and lowered FI and wt. The T-Orch group exhibited increased levels of ghrelin (3-fold), insulin, GH, blood levels of lipolysis products, TEE, and FI in addition to reduced glucose levels (P < 0.05). This group demonstrated no significant changes in wt. In the T-Sham group, T increased ghrelin and insulin levels (P < 0.05) with strong positive correlations (r = 0.663 and 0.644, respectively, P < 0.05), increased ATGL levels, RQ toward carbohydrate utilization ranges, and TEE, and reduced HSL levels (P < 0.05) with insignificant changes in FI or wt. Conclusions T administration in orchiectomized rats significantly increased orexigenic mediators such as ghrelin and insulin without inducing any significant changes in wt. The mechanism for this finding might be the increased TEE and the stimulation of lipolysis through the ATGL enzyme. The associated rise of GH might help in interference with accumulation of lipid in adipose tissue. Apart from the effect on GH, T-Sham showed similar effects of T supplementation.

Update on the synergistic effect of HSL and insulin in the treatment of metabolic disorders.

Hormone-sensitive lipase (HSL) is one of the three lipases in adipose tissue present during periods of energy demand. HSL is tightly controlled by insulin regulation via the central and peripheral systems. The suppressive effects of insulin on HSL are also associated with complex crosstalk with other pathways in the metabolic network. Because impaired insulin action is the driving force behind the pathogenesis of diabetes and other metabolic complications, elucidation of the intricate relationships between HSL and insulin may provide an in-depth understanding of these pandemic diseases and potentially identify strategies to inhibit disease development. Insulin not only differentially regulates HSL isoform transcription but also post-transcriptionally affects HSL phosphorylation by stimulating PKA and endothelin (ET-1), and controls its expression indirectly via regulating the activity of growth hormone (GH). In addition, a rapid elevation of HSL levels was detected after insulin injection in patients, which suggests that the inhibitory effects of insulin on HSL can be overridden by insulin-induced hypoglycemia. Conversely, individuals with hereditary HSL deficiency, and animals with experimental HSL deletion, showed major disruptions in mRNA/protein expression in insulin signaling pathways, ultimately leading to insulin resistance, diabetes, and fatty liver. Notably, HSL inactivation could cause insulin-independent fatty liver, while insulin resistance induced by HSL deficiency may further aggravate disease progression. The common beliefs that HSL is the overall rate-limiting enzyme in lipolysis and that insulin is an inhibitor of HSL have been challenged by recent discoveries; therefore, a renewed examination of their relationships is required. In this review, by analyzing current data related to the role of, and mutual regulation between, HSL and insulin and discussing unanswered questions and disparities in different lines of studies, the authors intend to shed light on our understanding of lipid metabolism and provide a rational basis for future research in drug development.

Effects of Japanese diet in combination with exercise on visceral fat accumulation.

In our previous study, we showed that among Japanese diets from different time periods, the 1975 Japanese diet has the greatest health benefits and is the most effective to prevent obesity. In addition, exercise is also effective to reduce obesity. Therefore, we conducted a human clinical trial combining the 1975 Japanese diet and exercise and, as a result, found a reduction in body weight, visceral fat, and serum lipids. However, the mechanism of this phenomenon was not determined. Therefore, in this study, we examined this mechanism in mice using a diet that was similar to that used in the human trial. The modern and 1975 Japanese diets were cooked, lyophilized, powdered, and fed freely to 5 wk old male C57 BL/6 J mice for 8 wk. In addition, the mice exercised on a treadmill. Total white adipose tissue weight decreased significantly due to the interaction between the 1975 Japanese diet and exercise. A histologic examination revealed that the hypertrophy of adipocytes was suppressed. To clarify this mechanism, the mRNA levels for lipid metabolism-related genes in epididymal adipose tissue were measured, and the mRNA level of hormone sensitive lipase (Hsl), which is related to lipolysis, was found to be significantly increased after intake of the 1975 Japanese diet combined with exercise. In the gut microbiota analysis, the Bacteroidetes to Firmicutes ratio, which is decreased in obese people, was increased by the 1975 Japanese diet and exercise. At the genus level, there was an increase in butyrate-producing bacteria as a result of the 1975 Japanese diet intake and exercise. A combination of the 1975 Japanese diet and exercise increased lipolysis in white adipose tissue and increased butyrate-producing bacteria in gut microbiota, and thereby suppressed fat accumulation.

Protective effects of N-acetyl-L-cysteine against binge drinking-induced fatty liver in mice

Objective: To investigate the roles of N-acetyl-L-cysteine (NAC) against binge drinking-induced fatty liver in mice. Methods: SPF male C57BL/6 mice were randomly divided into 3 groups, i.e. control group, model group, and NAC/ethanol group (n=10). Mice in model and NAC/ethanol groups were exposed to 3 doses of ethanol (6 g/kg bw) to induced fatty liver, while mice in control group received equal volume and equal energy of maltodextrin solution. NAC was administered to mice at 1 h before ethanol exposure (100 mg/kg bw, i.p.). The mice were sacrificed at 6 h after the last ethanol exposure. The liver and epididymal adipose tissues were collected. Histopathological examination and biochemical assay kit were used to evaluate the fat accumulation, while Western-blot was performed to detect the protein levels of some key factors involved in fat metabolism in liver and adipose tissues. Results: Compored with control group mice, the liver index and liver weight were significantly increased compared with model group, the liver index and TG level in NAC/ethanol group mice were all significantly decreased (P<0.05). Histological examination showed NAC effectively suppressed binge drinking-induced fat accumulation in mice liver. In addition, NAC had no significant effects on the protein levels of peroxisome proliferator-activated receptor-α (PPAR-α), Acy-CoA oxidase (ACOX), sterol regulatory element binding protein 1 c (SREBP-1c) and fatty acid synthase (FAS). Furthermore, the protein levels of hormone sensitive lipase (HSL) did not significantly differ among 3 groups, whereas NAC prevented binge drinking-induced increase of HSL phosphorylation at ser563 and ser660. Conclusion: NAC could effectively attenuate binge drinking-induced fatty liver, which might be associated with the inhibition of lipid mobilization by suppressing the phosphorylation of HSL.

Anti-hypertriglyceridemic Activity of Cornus Mas in Diabetic Rats

Introduction: Hypertriglyceridemia is among the multiple metabolic derangements seen in diabetes mellitus. Cornelian cherry (Cornus mas L.), belongs to the family Cornacea and has been shown to be helpful in treating hyperlipidemia. Aim: The study investigates mechanisms of action of dietary Cornelian Cherry fruit Dried Powder (CCDP) for ameliorating hypertriglyceridemia in diabetic rats. Materials and Methods: An experimental study with parallel controls was performed. Fifty six male adult rats were randomly assigned into 8 equal groups and treated as follows for 4 weeks. Negative control: Normal rats, basic diet; positive control: streptozotocin-induced diabetic rats, basic diet; T1 to T4 groups: diabetic rats fed with diets containing 0.25, 0.5, 1 and 2 g/ 100 g BW CCDP, respectively; T5: diabetic rats fed with basic diet plus 100 mg/kg BW of fenofibrate in drinking water and T6: normal rats fed with basic diet containing 1 g/100 g BW CCDP. Selected serum biochemical parameters, Lipoprotein Lipase (LPL) level as well as Adipose Tissue Hormone Sensitive Lipase (HSL) and Hepatic Peroxisome Proliferator-Activated Receptor Alpha (PPARα) levels were assayed. Analysis of data was performed by one-way ANOVA followed by Tukey’s test with p<0.05 as the significant level. Results: Treatment with CCDP at all dosages as well as fenofibrate decreased serum triglycerides and VLDL levels as compared to positive control. Serum AST significantly decreased in T1, T3 and T4 groups as compared to positive control rats. Serum LPL levels in the diabetic positive control group decreased significantly as compared to negative control. Rats in T1, T2, T4 and T5 groups showed increased serum LPL levels as compared to positive control. No significant difference was observed in hepatic PPARα levels among CCDP-treated and negative or positive controls. Positive control rats showed a significant decrease in adipose tissue HSL levels. Administration of CCDP in T3 group significantly increased HSL values as compared to positive control. CCDP in healthy rats did not change any of these parameters. Conclusion: Findings of the present study confirms positive effects of C. mas fruit on some liver function enzymes and hypertriglyceridemia and clarifies that increased serum LPL levels is at least partly responsible for hypotriglyceridemic effect of the fruit in rats with STZ-induced diabetes mellitus.

Ablation of systemic SIRT1 activity promotes nonalcoholic fatty liver disease by affecting liver-mesenteric adipose tissue fatty acid mobilization

Sirtuin 1 (SIRT1) has been reported to protect against nonalcoholic fatty liver disease (NAFLD) development. The mechanism of how SIRT1 deacetylase activity affects NAFLD has not been well investigated. The current investigation addressed the causal effect of systemic SIRT1 activity on NAFLD development and the underlying mechanism involved in both liver and mesenteric adipose tissue (MAT). Both SIRT1 homozygous mice ablated the catalytic activity (sirt1Y/Y) and their corresponding wild type littermates (WT) were fed a high fat diet (HFD, 60% calories from fat) for 34weeks. Sirt1Y/Y mice showed significantly higher level of hepatic triglyceride which was accompanied with higher levels of SREBP-1 and SCD1and decreased phosphorylation of LKB1 and AMPK in the liver. Compared with WT mice, mRNA expression of lipogenic genes (lxrα, srebp-1c, scd1 and fas) in the MAT increased significantly in sirt1Y/Y mice. Fatty acid oxidation biomarkers (acox1, acox3, cpt, ucp1, sirt3) in both liver and MAT were comparable between groups. Interestingly, we observed that in sirt1Y/Y mice, the mRNA level of hormone sensitive lipase (hsl), adipose triglyceride lipase (atgl) and perilipin-2 (plin-2), all involved in lipolysis, significantly increased in MAT, but not in epididymal adipose tissue. These changes positively correlated with circulating free fatty acid (FFA) concentrations and higher hepatic mRNA expression of cd36 for FFA uptake. The present study has provided novel evidence to suggest that under HFD-induced metabolic surplus, the lack of SIRT1 catalytic activity promotes release of FFA from MAT and escalate NAFLD by interfering with lipid homeostasis in both liver and MAT.

Ablation of systemic SIRT1 activity promotes nonalcoholic fatty liver disease by affecting liver‐mesenteric adipose tissue fatty acid mobilization

The incidence of nonalcoholic fatty liver disease (NAFLD) is escalating paralleled with obesity rates in both adults and children. Mammalian sirtuin 1 (SIRT1), a highly conserved NAD+‐dependent protein deacetylase, has been identified as a metabolic regulator of lipid homeostasis and a potential target for NAFLD prevention and treatment. However, the mechanism of how SIRT1, in particularly its deacetylase activity, affects NAFLD has not been well investigated. The current investigation addressed the causal effect of systemic SIRT1 activity on NAFLD development and the potential mechanism involved in both liver and mesenteric adipose tissue. At 6 weeks of age, both SIRT1 homozygous mice carrying a point mutation (H355Y, SIRT1Y/Y, n=10) that ablates the catalytic activity and their corresponding wild type littermates (WT, n=10) were fed with high fat diet (HFD, 60% calories from fat) for 34 weeks. Results showed no changes of body weight, liver weight and body composition (anlyzed by EchoMRI‐700) between WT and SIRT1Y/Y mice, but SIRT1Y/Y mice showed significantly higher level of hepatic triglyceride, and a trend of more severe hepatic steatosis by pathological analysis, which was accompanied with higher protein levels of LXRα, SREBP‐1, SCD1, involved in lipogenesis, and decreased phosphorylation of LKB1 and AMPK in the liver. In the mesenteric adipose tissue, mRNA expression of lipogenic genes (lxrα, srebp‐1c, scd1 and fas) increased significantly in SIRT1Y/Y mice, as compared with those of wild‐type mice. Fatty acid oxidation biomarkers (acox1, acox3, cpt, ucp1, sirt3) in both liver and mesenteric adipose tissue were comparable between groups. Interestingly, we observed that in SIRT1Y/Y mice, the mRNA level of hormone sensitive lipase (hsl), adipose triglyceride lipase (atgl) and perilipin‐2, involved in lipolysis, significantly increased in mesenteric adipose tissue (not in epididymal adipose tissue), as compared with those of WT. These changes were correlated with higher circulating free fatty acid (FFA) concentrations and higher hepatic mRNA expression of cd36 for FFA uptake. Taken together, the present study provided novel experimental evidence that under HFD‐induced metabolic surplus, the lack of SIRT1 catalytic activity promoted release of FFA from mesenteric fat into systemic circulation and escalated NAFLD by interfering with lipid homeostasis in both liver and mesenteric adipose tissue.Support or Funding InformationSupported by USDA/NIFA (2015‐67017) and USDA/ARS (58‐1950‐0014)

Abstract 254: A Novel Mechanism for Aberrant Aldosterone Production in Heart Failure

Objective: Aldosterone production by the adrenal gland is mainly regulated by the renin-angiotensin system (RAS), which is normally suppressed by a high-salt diet. However, plasma aldosterone levels are elevated in proportion to the severity of heart failure even if caused by high-salt intake. The inappropriate increase in plasma aldosterone level could not be diminished by pharmacological blockade of the RAS. We investigated the cellular mechanism underlying the aberrant aldosterone production in heart failure beyond the activation of RAS. Methods and Results: Dahl-salt sensitive rats fed high salt diet developed malignant hypertension, resulting in heart failure with increased plasma aldosterone level. The gene expression pattern of the adrenal gland from heart failure rats showed strikingly upregulated β3-adrenergic receptor (β3-AR) expression. Immunohistochemical staining confirmed that heart failure increased β3-AR expression in zona glomerulosa cells in adrenal gland of Dahl-salt sensitive rats. And the staining of pimonidazole, a hypoxia probe, revealed that the outer layer of adrenal cortex in the heart failure rat was under hypoxic condition. Hypoxia and DMOG, a chemical stabilizer of hypoxia-inducible factor, strongly induced β3-AR mRNA expression in H295R human adrenocortical carcinoma cells. Hypoxia-reoxygenation stimuli induced aldosterone production from H295R cells, which was blocked by SR59230A, a selective β3-AR antagonist, but not by losartan, an angiotensin II receptor blocker (ARB). Hypoxia endowed catecholamine responsiveness in H295R cells by increased β3-AR expression, wherein β3-AR agonist increased the phosphorylation levels of hormone-sensitive lipase (HSL) through activation of ERK pathway. Conclusions: In the present study, we demonstrated that the β3-AR induction in adrenal gland by hypoxia played an important role in aberrant aldosterone production in heart failure. The catecholamine stimuli through β3-AR in adrenocortical cells caused the ERK-mediated phosphorylation of HSL, suggesting that the HSL promotes lipolysis to supply substrates for the aldosterone biosynthesis.

Different effects of growth hormone and fasting on the induction patterns of two hormone-sensitive lipase genes in red seabream Pagrus major

Growth hormone (GH) increases phosphorylation and mRNA levels of hormone-sensitive lipase (HSL) in the livers of some marine teleosts. The hepatic GH-HSL axis appears to play important roles in fasting-induced lipolysis. However, it is not known whether GH exerts similar effects on HSL in fish adipose tissues. Functional differentiation of two fish-specific HSL isoforms (HSL1 and HSL2) also remains unclear. The present study seeks to address two unanswered questions about fish lipolysis using red seabream (Pagrus major): (1) Does GH increase phosphorylation and mRNA levels of HSL in adipose tissue? (2) How do GH and fasting affect mRNA levels of two HSL isoform genes in the liver and adipose tissue? To this end, we first cloned HSL1 and HSL2 cDNAs and investigated their tissue distribution. Transcripts of both HSLs and HSL1 proteins were abundant in the visceral adipose tissue, gonads, and liver, suggesting the important role of HSL in adipose tissue lipolysis. HSL2 transcript levels were 20–65% those of HSL1 except in the skin, and HSL2 proteins were not detected by our in-house antisera. Ex vivo administration of GH increased HSL1 phosphorylation, non-esterified fatty acid (NEFA) release, and levels of HSL1 and HSL2 mRNA in both the liver and visceral adipose tissue. Hepatic HSL2 mRNA was particularly sensitive to GH administration and sometimes exceeded HSL1 mRNA levels with up to 13-fold induction. In contrast, fasting for 4 and 7d increased HSL1 mRNA levels, but had only marginal effects on HSL2 mRNA levels in both adipose tissue or liver. We concluded that GH would increase HSL mRNAs during adipose tissue lipolysis in red seabream; however, GH and fasting result in different induction ratio of two HSL isoform genes, suggesting that other hormone(s) also contributes to fasting-induced lipolysis.

Enhanced activity of hormone sensitive lipase (HSL) in mesenteric but not epididymal fat correlates with higher production of epinephrine in mesenteric adipocytes in rat model of cachectic rheumatoid arthritis

Cachectic rheumatoid arthritis, the less frequent form of the disease, is associated with loss of fat mass and often more severe course of the disease. Its experimental model represents rat adjuvant arthritis (AA) characterized by edema, lack of appetite, sharp body weight and fat loss. As individual fat depots display functional differences, here we studied lipolytic activity and sensitivity to lipolytic stimuli of nodeless epididymal fat (eWAT) and perinodal mesenteric fat (mWAT) depots at the peak of AA. We also examined changes in catecholamine and cytokine levels involved in lipolysis in plasma and/or isolated adipocytes from both WATs to identify the contribution of local, adipocyte-based processes and/or systemic events to adiposity loss in cachectic rheumatoid arthritis. AA was induced to male Lewis rats by complete Freund’s adjuvant. Groups of ad libitum-fed and pair-fed controls were used to distinguish the effects of food restriction from inflammation-induced cachexia. Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and its phosphorylated form (pHSL) were analyzed by western blot. CRP and catecholamine levels in plasma or adipocyte lysates were determined using ELISA kits. Cytokine-induced neutrophil chemoattractant-1 (CINC-1/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), IL-1β, IL-6, IL-10 and leptin in adipocyte lysate were analyzed by quantitative protein microarray. Plasma glycerol and FFA were measured spectrophotometrically. AA rats developed severe cachexia, with lower adiposity in mWAT compared to normal and pair-fed controls, whereas in eWAT the adiposity was similarly reduced in AA and pair-fed groups. ATGL levels in both WATs were not affected by AA or pair feeding. AA upregulated levels of HSL, pHSL and pHSL/HSL ratio in mWAT, whereas none of these parameters has changed in eWAT of AA rats or in either WATs of pair-fed rats. In AA rats plasma glycerol was elevated, whereas FFA concentration was reduced. Plasma norepinephrine and epinephrine were increased in AA compared with both groups of controls. In eWAT adipocytes, AA but not pair feeding, upregulated norepinephrine levels. In mWAT adipocytes, AA rats showed higher epinephrine levels than pair-fed controls. Leptin levels in both WATs were depleted in AA animals in accordance with body weight loss. None of the measured cytokines in eWAT and mWAT was enhanced. Our results demonstrate augmented lipolytic activity in mWAT and not eWAT during cachectic arthritis. The adipocyte-derived cytokines do not seem to contribute to activated lipolysis. We first demonstrated enhanced presence of norepinephrine in perinodal adipocytes that may contribute to the regulation of local lipolytic activity by auto/paracrine fashion and thus provide independent fuel supply to activated lymph nodes.

A polyphenol-rich fraction obtained from table grapes decreases adiposity, insulin resistance and markers of inflammation and impacts gut microbiota in high-fat-fed mice

The objective of this study was to determine if consuming an extractable or nonextractable fraction of table grapes reduced the metabolic consequences of consuming a high-fat, American-type diet. Male C57BL/6J mice were fed a low fat (LF) diet, a high fat (HF) diet, or an HF diet containing whole table grape powder (5% w/w), an extractable, polyphenol-rich (HF-EP) fraction, a nonextractable, polyphenol-poor (HF-NEP) fraction or equal combinations of both fractions (HF-EP+NEP) from grape powder for 16weeks. Mice fed the HF-EP and HF-EP+NEP diets had lower percentages of body fat and amounts of white adipose tissue (WAT) and improved glucose tolerance compared to the HF-fed controls. Mice fed the HF-EP+NEP diet had lower liver weights and triglyceride (TG) levels compared to the HF-fed controls. Mice fed the HF-EP+NEP diets had higher hepatic mRNA levels of hormone sensitive lipase and adipose TG lipase, and decreased expression of c-reactive protein compared to the HF-fed controls. In epididymal (visceral) WAT, the expression levels of several inflammatory genes were lower in mice fed the HF-EP and HF-EP+NEP diets compared to the HF-fed controls. Mice fed the HF diets had increased myeloperoxidase activity and impaired localization of the tight junction protein zonula occludens-1 in ileal mucosa compared to the HF-EP and HF-NEP diets. Several of these treatment effects were associated with alterations in gut bacterial community structure. Collectively, these data demonstrate that the polyphenol-rich, EP fraction from table grapes attenuated many of the adverse health consequences associated with consuming an HF diet.

Proteomic identification of fat-browning markers in cultured white adipocytes treated with curcumin.

We previously reported that curcumin induces browning of primary white adipocytes via enhanced expression of brown adipocyte-specific genes. In this study, we attempted to identify target proteins responsible for this fat-browning effect by analyzing proteomic changes in cultured white adipocytes in response to curcumin treatment. To elucidate the role of curcumin in fat-browning, we conducted comparative proteomic analysis of primary adipocytes between control and curcumin-treated cells using two-dimensional electrophoresis combined with MALDI-TOF-MS. We also investigated fatty acid metabolic targets, mitochondrial biogenesis, and fat-browning-associated proteins using combined proteomic and network analyses. Proteomic analysis revealed that 58 protein spots from a total of 325 matched spots showed differential expression between control and curcumin-treated adipocytes. Using network analysis, most of the identified proteins were proven to be involved in various metabolic and cellular processes based on the PANTHER classification system. One of the most striking findings is that hormone-sensitive lipase (HSL) was highly correlated with main browning markers based on the STRING database. HSL and two browning markers (UCP1, PGC-1α) were co-immunoprecipitated with these markers, suggesting that HSL possibly plays a role in fat-browning of white adipocytes. Our results suggest that curcumin increased HSL levels and other browning-specific markers, suggesting its possible role in augmentation of lipolysis and suppression of lipogenesis by trans-differentiation from white adipocytes into brown adipocytes (beige).

CTRP1 protects against diet-induced hyperglycemia by enhancing glycolysis and fatty acid oxidation

Complement-C1q/tumor necrosis factor-α related protein 1 (CTRP1) is a 35-kDa glycoprotein that is secreted from various tissues. Although CTRP1 is highly increased in patients with type II diabetes and obesity, the metabolic roles of CTRP1 remain largely unknown. To unveil the physiological roles of CTRP1 in vivo, CTRP1 transgenic (TG) mice were challenged by a high-fat diet (HFD) and a high-sucrose drink (HS). Homeostatic model assessment-estimated insulin resistance values were decreased in HFD- or HS-fed CTRP1 TG mice compared with wild-type control mice. In this context, CTRP1 stimulated glucose uptake through the glucose transporter GLUT4 translocation to the plasma membrane and also increased glucose consumption by stimulating glycolysis. To analyze the roles of CTRP1 in lipid metabolism, acetyl-CoA carboxylase (ACC) and hormone-sensitive lipase levels were determined in CTRP1 TG mice, and the effect of CTRP1 on fatty acid oxidation was assessed in C2C12 myotubes. CTRP1 was found to inhibit ACC by phosphorylation and to stimulate fatty acid oxidation in C2C12 myotubes. Taken together, CTRP1 performs active catabolic roles in vivo. Therefore, CTRP1 seems to perform a defensive function against nutritional challenges.

Habitual exercise training acts as a physiological stimulator for constant activation of lipolytic enzymes in rat primary white adipocytes

It is widely accepted that lipolysis in adipocytes are regulated through the enzymatic activation of both hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) via their phosphorylation events. Accumulated evidence shows that habitual exercise training (HE) enhances the lipolytic response in primary white adipocytes with changes in the subcellular localization of lipolytic molecules. However, no study has focused on the effect that HE exerts on the phosphorylation of both HSL and ATGL in primary white adipocytes. It has been shown that the translocation of HSL from the cytosol to lipid droplet surfaces requires its phosphorylation at Ser-563. In primary white adipocytes obtained from HE rats, the level of HSL and ATGL proteins was higher than that in primary white adipocytes obtained from sedentary control (SC) rats. In HE rats, the level of phosphorylated ATGL and HSL was also significantly elevated compared with that in SC rats. These differences were confirmed by Phos-tag SDS-PAGE, a technique used to measure the amount of total phosphorylated proteins. Our results suggest that HE can consistently increase the activity of both lipases, thereby enhancing the lipolysis in white fat cells. Thus, HE helps in the prevention and treatment of obesity-related diseases by enhancing the lipolytic capacity.

Green tea catechins enhance norepinephrine-induced lipolysis via a protein kinase A-dependent pathway in adipocytes

Green tea catechins have been shown to attenuate obesity in animals and humans. The catechins activate adenosine monophosphate-activated protein kinase (AMPK), and thereby increase fatty acid oxidation in liver and skeletal muscles. Green tea catechins have also been shown to reduce body fat in humans. However, the effect of the catechins on lipolysis in adipose tissue has not been fully understood. The aim of this study was to clarify the effect of green tea catechins on lipolysis in adipocytes and to elucidate the underlying mechanism. Differentiated mouse adipocyte cell line (3T3-L1) was stimulated with green tea catechins in the presence or absence of norepinephrine. Glycerol and free fatty acids in the media were measured. Phosphorylation of hormone-sensitive lipase (HSL) was determined by Western blotting, and the mRNA expression levels of HSL, adipose triglyceride lipase (ATGL), and perilipin were determined by quantitative RT-PCR. The cells were treated with inhibitors of protein kinase A (PKA), protein kinase C (PKC), protein kinase G (PKG), or mitogen-activated protein kinase (MAPK) to determine the responsible pathway. Treatment of 3T3-L1 adipocytes with green tea catechins increased the level of glycerol and free fatty acids released into the media in the presence, but not absence, of norepinephrine, and increased the level of phosphorylated HSL in the cells. The catechins also increased mRNA and protein levels of HSL and ATGL. PKA inhibitor (H89) attenuated the catechin-induced increase in glycerol release and HSL phosphorylation. The results demonstrate that green tea catechins enhance lipolysis in the presence of norepinephrine via a PKA-dependent pathway in 3T3-L1 adipocytes, providing a potential mechanism by which green tea catechins could reduce body fat.