The Expression of TBC1 Domain Family, Member 4 (TBC1D4) in Skeletal Muscles of Insulin-Resistant Mice in Response to Sulforaphane

The Expression of TBC1 Domain Family, member 4 (TBC1D4) in Skeletal Muscles of Insulin-Resistant Mice in Response to Sulforaphane. Background: Obesity is commonly accompanied by impaired glucose homeostasis. Decreased glucose transport to the peripheral tissues, mainly skeletal muscle, leads to reduced total glucose disposal and hyperglycemia. TBC1D4 gene is involved in the trafficking of GLUT4 to the outer cell membrane in skeletal muscle. Sulforaphane (SFN) has been suggested as a new potential anti-diabetic compound acting by reducing blood glucose levels through mechanisms not fully understood (1). The aim of this study is to investigate the effects SFN on TBC1D4 and GLUT4 gene expression in skeletal muscles of DIO mice, in order to elucidate the mechanism(s) through which SFN improves glucose homeostasis. Methodology: C57BL/6 mice (n=20) were fed with a high fat diet (60%) for 16 weeks to generate diet induced obese (DIO) mice with body weights between 45–50 gm. Thereafter, DIO mice received either SFN (5mg/kg BW) (n=10) or vehicle (n=10) as controls daily by intraperitoneal injections for four weeks. Glucose tolerance test (1g/kg BW, IP) and insulin sensitivity test (ITT) were conducted (1 IU insulin/ g BW, IP route) at the beginning and end of the third week of the injection. At the end of 4 weeks of the injection, samples of blood and skeletal muscles of both hindlimbs were collected. The expression levels of GLUT4 and TBC1D4 genes were analyzed by qRT-PCR. Blood was also used for glucose, adiponectin and insulin measurements. Results: SFN-treated DIO mice had significantly lower non-fasting blood glucose levels than vehicle-treated mice (194.16 ± 14.12 vs. 147.44 ± 20.31 mg/dL, vehicle vs. SFN, p value=0.0003). Furthermore, GTT results indicate that the blood glucose levels at 120 minutes after glucose infusion in was (199.83±34.53 mg/dl vs. 138.55±221.78 mg/dl) for vehicle vs. SFN with p=0.0011 respectively. ITT showed that SFN treatment did not enhance insulin sensitivity in DIO mice. Additionally, SFN treatment did not significantly change the expression of TBC1D4, and GLUT4 genes in skeletal muscles compared to vehicle treatment (p values >0.05). Furthermore, SFN treatment did not significantly affect the systemic insulin (1.84±0.74 vs 1.54±0.55 ng/ml, p=0.436), or adiponectin (11.96 ±2.29 vs 14.4±3.33 ug/ml, p=0.551) levels in SFN vs. vehicle-treated DIO mice, respectively. Conclusion: SFN treatment improves glucose disposal in DIO mice, which is not linked to the gene expression of GLUT4 and TBC1D4 and its mechanism of glucose disposal in skeletal muscles. Furthermore, SFN treatment did not improve insulin level, and the insulin sensitizer hormone adiponectin as potential players for enhancing insulin sensitivity.QNRF-NPR

PROFILE OF OXIDATIVE STRESS GENES IN RESPONSE TO OBESITY TREATMENT

Background: Oxidative stress (OS) is an imbalance between free radical production and the antioxidants defense in the body. Previous studies demonstrated the correlation of OS to the increased risk of developing metabolic disorders such as obesity. Sulforaphane (SFN), a bioactive compound, can protect against inflammation and OS, thus an effective anti-obesity supplement. Aim: This study explores the impact of SNF on OS in diet induced obese (DIO) mice via profiling of OS genes and pathways in skeletal muscles related to the anti-obesity effect. Methods: Wild-type CD1 male mice and the knockout of nuclear factor (erythroid-derived 2) like 2 (NrF2) mice were fed a high-fat diet (HFD) for 16 weeks; to induce obesity. Subsequently, each group was subdivided into two subgroups and received either Vehicle (25?l) or SFN (5 mg/kg BW) for four weeks. Body weight was measured daily, and a glucose tolerance test (GTT) was performed after 21 days of treatment. Afterward, mice were decapitated, blood and tissue samples were collected and snap-frozen immediately. Total RNA was extracted from Skeletal muscle and epididymal white adipose tissue (eWAT), leptin expression was measured in (eWAT), and 84 OS genes in skeletal muscle were examined using RT-PCR. Results: Significant reduction in body weight in SFN treated WT mice, while no change in KO mice. Plasma glucose, leptin, and leptin gene expression (eWAT) were significantly reduced in the WT-DIO SFN treated group, while no changes were detected in KO mice. SFN decreases OS damage in skeletal muscles, such as lipid peroxidation and production of reactive oxygen species (ROS). Conclusion: This study demonstrated that SFN had lowered body weight in WT-DIO mice by decreasing OS damage in skeletal muscles through the NrF2 pathway and can be a potential anti-obesity drug

The AlignMend Band

Proper long-distance running technique not only minimizes chance of injury, but it also allows runners to be more efficient. When proper form is practiced and unnecessary movement is eliminated, more of the runner’s energy is transferred to propulsion, moving the runner forward, thus maximizing efficiency. There is plenty of research on lower body form, for prevention of joint and knee injuries; however, since there is little research and barely any products that look at the remainder of the body, we will focus on correcting the long-distance runner’s improper upper body form. The hands control the tension in the upper body, while the swing of the arms works in conjunction with leg stride to move the runners forward as well as maintain cadence and rhythm. When the runner’s arms are flailing up and down or side-to-side, it is more likely for him/her to slouch and not breathe as efficiently. If the runner holds his/her hands too high up to his/her chest, s/he will feel tightness and tension in his/her shoulders and neck. Analysis of runners’ gait (pattern of movement when walking), specifically of the upper body, can help avoid injury and improve efficiency. This project aims to design a portable device, called “The AlignMend Band”, that specifically monitors the movement of a runner’s arms and provides gait information. When the runner’s arms move outside of the angle specifications for proper long-distance running form, the device will alert him/ her with a vibration to adjust his/her upper body form and continue doing so until proper form is achieved. The AlignMend Band acts as a standalone device and is expected to greatly improve the efficiency of long-distance runners

The AlignMend Band

Proper long-distance running technique not only minimizes chance of injury, but it also allows runners to be more efficient. When proper form is practiced and unnecessary movement is eliminated, more of the runner’s energy is transferred to propulsion, moving the runner forward, thus maximizing efficiency. There is plenty of research on lower body form, for prevention of joint and knee injuries; however, since there is little research and barely any products that look at the remainder of the body, we will focus on correcting the long-distance runner’s improper upper body form. The hands control the tension in the upper body, while the swing of the arms works in conjunction with leg stride to move the runners forward as well as maintain cadence and rhythm. When the runner’s arms are flailing up and down or side-to-side, it is more likely for him/her to slouch and not breathe as efficiently. If the runner holds his/her hands too high up to his/her chest, s/he will feel tightness and tension in his/her shoulders and neck. Analysis of runners’ gait (pattern of movement when walking), specifically of the upper body, can help avoid injury and improve efficiency. This project aims to design a portable device, called “The AlignMend Band”, that specifically monitors the movement of a runner’s arms and provides gait information. When the runner’s arms move outside of the angle specifications for proper long-distance running form, the device will alert him/ her with a vibration to adjust his/her upper body form and continue doing so until proper form is achieved. The AlignMend Band acts as a standalone device and is expected to greatly improve the efficiency of long-distance runners

Early Hepatic Gene Expression Profile of Lipid Metabolism of Mice on High Fat Diet after Treatment with Anti‐Obesity Drugs

Obesity is a multifactorial disorder of global scale. The liver plays a vital role in fat metabolism. Disorder of hepatic fat metabolism is associated with obesity and fatty liver disease. This study aimed to detect the effects of anti‐obesity drugs (sulforaphane; SFN and leptin) on hepatic gene expression of fat metabolism in mice that were fed HFD during an early time of DIO. Thirty‐two wild type (WT) ten‐week‐old CD1 male mice were fed high fat diet for four weeks in order to induce diet‐induced obesity (DIO). The mice were treated with a vehicle, or SFN for one week and then each group is treated with leptin or saline for 24 hours. Four groups of treatment were obtained; control group (vehicle + saline), group 2 (vehicle + leptin), group 3 (SFN + saline), and group 4 (SFN + leptin). Body weight and food intake were monitored during the treatment period. Following the treatments, liver tissue was collected, and total RNA was extracted to assess the expression of 84 genes involved in hepatic fat metabolism using RT‐PCR profiler array technique. Leptin treatment upregulated the genes involved in fatty acid beta‐oxidation (Acsbg2, Acsm4) and fatty acyl‐CoA biosynthesis (Acot6, Acsl6), and down‐regulated the fatty acid transport gene (Slc27a2). SFN upregulated acyl‐CoA hydrolase (Acot3) and long chain fatty acid activation for lipids synthesis and beta‐oxidation (Acsl1). Leptin + SFN upregulated fatty acid beta‐oxidation‐related genes (Acad11, Acam) and acyl‐CoA hydrolases (Acot3, Acot7), and downregulated fatty acid elongation gene, Acot2. As a result, treatment with both SFN and leptin has a more profound effect in ameliorating the pathways involved in hepatic lipogenesis and TG accumulation and body weight gain than other types of intervention. We conclude that early intervention of obesity pathogenesis could ameliorate the metabolic changes of fat metabolism in the liver, as observed in mice on HFD in response to SFN anti‐obesity treatment.NPRP Grant No. 9‐351‐3‐07

Sulforaphane Downregulates Hepatic Fibroblast Growth Factor 21 (FGF21) of Diet Induced Obese Mice

Background: Fibroblast growth factor 21 is a hormone-like protein that plays a critical role as an energy regulator. Sulforaphane (SFN) is expected to have potential therapeutic effects in treating obesity. This study aims to investigate the effect of SFN treatment on hepatic gene expression of FGF-21 of diet induced obese mice. Methods: CD1 male mice and two groups of lean and diet induced obesity (DIO) model after feeding a high fat diet were used. Afterward, both lean and DIO mice were treated for four weeks with either SFN (5mg/kg BW) (n=10) or Vehicle (n=10). After that, blood and liver samples were collected and analyzed. Hepatic FGF-21 gene expression was measured using qRT-PCR. Results: Treatment of DIO mice with SFN causes a significant reduction in body weight gain (15.42%) compared to DIO-vehicle group, which showed a weight gain by (3.86%), p-value<0.0001. In addition, SFN treatment to lean group did not affect body weight. DIO-SFN treated mice showed a significant reduction in fasting glucose, leptin, and insulin levels compared to DIO-vehicle treated group, p-value<0.05. Hepatic FGF-21 gene expression was significantly upregulated in DIO-vehicle compared to lean-vehicle mice with ˜ 3 folds, p-value<0.05. Treatment of DIO with SFN causes a significant downregulation of FGF-21 gene expression by ˜9 folds compared with DIO-vehicle treated group, p-value<0.05. Conclusions: Treatment of DIO mice with SFN causes downregulation of hepatic FGF21 expression in obese mice. The effects of SFN on FGF21 gene expression could be a direct effect or secondary to weight loss, which warrants further studies.QNRF, NPRP 9 -351-3-07

Sulforaphane reduces obesity by reversing leptin resistance.

The ascending prevalence of obesity in recent decades is commonly associated with soaring morbidity and mortality rates, resulting in increased health-care costs and decreased quality of life. A systemic state of stress characterized by low-grade inflammation and pathological formation of reactive oxygen species (ROS) usually manifests in obesity. The transcription factor nuclear factor erythroid-derived 2-like 2 (NRF2) is the master regulator of the redox homeostasis and plays a critical role in the resolution of inflammation. Here, we show that the natural isothiocyanate and potent NRF2 activator sulforaphane reverses diet-induced obesity through a predominantly, but not exclusively, NRF2-dependent mechanism that requires a functional leptin receptor signaling and hyperleptinemia. Sulforaphane does not reduce the body weight or food intake of lean mice but induces an anorectic response when coadministered with exogenous leptin. Leptin-deficient mice and leptin receptor mutant mice display resistance to the weight-reducing effect of sulforaphane, supporting the conclusion that the antiobesity effect of sulforaphane requires functional leptin receptor signaling. Furthermore, our results suggest the skeletal muscle as the most notable site of action of sulforaphane whose peripheral NRF2 action signals to alleviate leptin resistance. Transcriptional profiling of six major metabolically relevant tissues highlights that sulforaphane suppresses fatty acid synthesis while promoting ribosome biogenesis, reducing ROS accumulation, and resolving inflammation, therefore representing a unique transcriptional program that leads to protection from obesity. Our findings argue for clinical evaluation of sulforaphane for weight loss and obesity-associated metabolic disorders.Qatar National Research Fund (NPRP9-351-3-075