The role of AMPK/mTOR/S6K1 signaling axis in mediating the physiological process of exercise-induced insulin sensitization in skeletal muscle of C57BL/6 mice

AbstractThe crosstalk between mTORC1/S6K1 signaling and AMPK is emerging as a powerful and highly regulated way to gauge cellular energy and nutrient content. The aim of the current study was to determine the mechanism by which exercise training reverses lipid-induced insulin resistance and the role of AMPK/mTOR/S6K1 signaling axis in mediating this response in skeletal muscle. Our results showed that high-fat feeding resulted in decreased glucose tolerance, which was associated with decreased Akt expression and increased intramuscular triglyceride deposition in the skeletal muscle of C57BL/6 mice. Impairments in lipid metabolism were accompanied by increased total protein and phosphorylation of S6K1, SREBP-1c cleavage, and decreased AMPK phosphorylation. Exercise training reversed these impairments, resulting in improved serum lipid profiles and glucose tolerance. C2C12 myotubes were exposed to palmitate, resulting in an increased insulin-dependent Akt Ser473 phosphorylation, associated with a significant increase in the level of phosphorylation of S6K1 on T389. All these changes were reversed by activation of AMPK. Consistent with this, inhibition of AMPK by compound C induced an enhanced phosphorylation of both S6K1 and Akt, and silencing of S6K1 with siRNA showed no effect on Akt phosphorylation in both the absence and presence of palmitate cultured myotubes. In addition, compound C led to an elevated SREBP-1c cleavage but was blocked by S6K1 siRNA. In summary, exercise training inhibits SREBP-1c cleavage through AMPK/mTOR/S6K1 signaling, resulting in decreased intramyocellular lipid accumulation. Our results provide new insights into the mechanism by which AMPK/mTOR/S6K1 signaling axis mediates the physiological process of exercise-induced insulin sensitization

PO-193 Exercise training decreased lipid accumulation in murine skeletal muscle through Sestrin2-mediated SHIP2-JNK signaling pathway

Objective Obesity is becoming increasingly prevalent and is an important contributor to the worldwide burden of diseases. It is widely accepted that exercise training is beneficial for the prevention and treatment of obesity. However, the underlying mechanism by which exercise training improving skeletal muscle lipid metabolism is still not fully described. Sestrins (Sestrin1-3) are highly conserved stress-inducible protein. Concomitant ablation of Sestrin2 and Sestrin3 has been reported to provoke hepatic mTORC1/S6K1 activation and insulin resistance even without nutritional overload and obesity, implicating that Sestrin2 and Sestrin3 have an important homeostatic function in the control of mammalian glucose and lipid metabolism. Our previous results demonstrated that physical exercise increased Sestrin2 expression in murine skeletal muscle, while the role of Sestrin2 in regulating lipid metabolism remains unknown.  SH2 domain containing inositol 5-phosphatase (SHIP2) acts as a negative regulator of the insulin signaling both in vitro and in vivo. An increased expression of SHIP2 inhibits the insulin-induced Akt activation, glucose uptake, and glycogen synthesis in 3T3-L1 adipocytes, L6 myotubes and tissues of animal models. Alterations of SHIP2 expression and/or enzymatic function appear to have a profound impact on the development of insulin resistance. However, the regulatory function of SHIP2 in lipid metabolism after exercise remains unclear. It has been reported that SHIP2 modulated lipid metabolism through regulating the activity of c-Jun N-terminal kinase (JNK) and Sterol regulatory element-binding protein-1 (SREBP-1). JNK is a subclass of mitogen-activated protein kinase (MAPK) signaling pathway in mammalian cells and plays a crucial role in metabolic changes and inflammation associated with a high-fat diet. Inhibition of JNK reduces lipid deposition and proteins level of fatty acid de novo synthesis in liver cells. It has been reported that Sestrin2 regulated the phosphorylation of JNK, however the underlying mechanism remains unclear. SREBP-1 is important in regulating cholesterol biosynthesis and uptake and fatty acid biosynthesis, and SREBP-1 expression produces two different isoforms, SREBP-1a and SREBP-1c. SREBP-1c is responsible for regulating the genes required for de novo lipogenesis and its expression is regulated by insulin. SREBP-1a regulates genes related to lipid and cholesterol production and its activity is regulated by sterol levels in the cell. Altogether, the purpose of this study was to explore the effect and underlying mechanism of Sestrin2 on lipid accumulation after exercise training. Methods Male wild type and SESN2−/− mice were divided into normal chow (NC) and high-fat diet (HFD) groups to create insulin resistance mice model. After 8 weeks the IR model group was then divided into HFD sedentary control and HFD exercise groups (HE). Mice in HE group underwent 6-week treadmill exercise to reveal the effect of exercise training on lipid metabolism in insulin resistance model induced by HFD. We explored the mechanism through which Sestrin2 regulated lipid metabolism in vitro by supplying palmitate, overexpressing or inhibiting SESNs, SHIP2 and JNK in myotubes. Results We found that 6-week exercise training decreased body weight, BMI and fat mass in wild type and SESN2-/- mice after high-fat diet (HFD) feeding. And exercise training decreased the level of plasma glucose, serum insulin, triglycerides and free fatty acids in wild type but not in Sestrin2-/- mice. Lipid droplet in skeletal muscle was also decreased in wild type but did not in Sestrin2-/- mice. Moreover, exercise training increased the proteins expression involved in fatty acid oxidation and decreased the proteins which related to fatty acid de novo synthesis. The results of oil red staining and the change of proteins related to fatty acid de novo synthesis and beta oxidation in myotubes treated with palmitate, Ad-SESN2 and siRNA-Sestrin2 were consisted with the results in vivo, which suggested that Sestrin2 was a key regulator in lipid metabolism. Exercise training increased Sestrin2 expression and reversed up-regulation of SHIP2 and pJNK induced by HFD in wild type mice but not in Sestrin2-/- mice. In parallel, overexpression of Sestrin2 decreased the level of SHIP2 and pJNK induced by palmitate while Sestrin2 knock down by siRNA-Sestrin2 treatment did not change the expression of SHIP2 and pJNK, which suggested that Sestrin2 modulated SHIP2 and JNK in the state of abnormal lipid metabolism. Inhibition of SHIP2 reduced the activity of JNK, increased lipid accumulation and the proteins of fatty acid synthesis after palmitate treatment and over expression of Sestrin2, which suggest that Sestrin2 modulated lipid metabolism through SHIP2/JNK pathway. Conclusions Sestrin2 plays an important role in improving lipid metabolism after exercise training, and Sestrin2 regulates lipid metabolism by SHIP2-JNK pathway in skeletal muscle

Mapping of nighttime light trends and refugee population changes in Ukraine during the Russian–Ukrainian War

The nighttime lights accurately and coherently depict how humans live. This study uses nighttime light measurements to quantify changes in nighttime lighting and refugee population in Ukraine before and after the war. We combined the Theil–Sen estimator with the M-K test to explore the trends of nighttime light. In addition, we constructed a linear model using nighttime light data and a portion of the UNHCR refugee data. Our results reveal that 1 week after the start of the Russo-Ukrainian War, the nighttime light area and the average nighttime light DN value in Ukraine exhibited a steep decline of about 50 percent. Our findings showed taht refugee population changes calculated through models and nighttime light data were mostly consistent with UNHCR data. We thought that the nighttime light data might be used directly to dynamically estimate changes in the refugee movement throughout the war. Nighttime light changes has significant implications for international humanitarian assistance and post-war reconstruction

Exercise improves lipid metabolism disorders induced by high-fat diet in a SESN2/JNK- independent manner

SESN2 and JNK are emerging powerful stress-inducible proteins in regulating lipid metabolism. The aim of this study was to determine the underlying mechanism of SESN2/JNK signaling in exercise improving lipid disorder induced by high-fat diet (HFD). Our data showed that HFD and SESN2 knockout resulted in abnormalities including elevated body weight, increased fat mass, serum total cholesterol (TC), lipid biosynthesis related proteins, and a concomitant increase of pJNK-Thr183/Tyr185. The above changes were reversed by exercise training. SESN2 silencing or JNK inhibition in palmitate-treated C2C12 further confirmed that SESN2 and JNK play a vital role in lipid biosynthesis. Rescue experiment further demonstrated that SESN2 reduced lipid biosynthesis through inhibition of JNK. SESN2/JNK signaling axis regulates lipid biosynthesis in both animal and cell models with abnormalities of lipid metabolism induced by HFD or palmitate treatment. This study provided evidence that exercise ameliorated lipid metabolic disorder induced by HFD feeding or by SESN2 knockout. SESN2 may improve lipid metabolism through inhibition JNK expression in skeletal muscle cells, providing a molecular mechanism that may represent an attractive target for the treatment of lipid disorder. Novelty: ● Exercise improved lipid disorder induced by HFD feeding and SESN2 knockout. ● SESN2 and JNK play a vital role in lipid biosynthesis in vivo and in vitro. ● SESN2 suppressed JNK to improve lipid metabolism in skeletal muscle cells.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

SESN2 ablation weakens exercise benefits on resilience of gut microbiota following high-fat diet consumption in mice

Gut dysbiosis is associated with several pathological processes. Previous study showed that regular exercise can protect against dysmetabolism in high-fat diet (HFD) fed mice through butyrate-SESN2 pathway, and SESN2 ablation weakened the protective effects of exercise. Here, we investigated whether SESN2-deficiency suppresses the exercise response to microbiota composition and subsequently reduces the benefits of exercise on dysmetabolism induced by HFD. Wild type (WT) and SESN2−/− mice were assigned to five-groups, fed with either normal chow or HFD and with or without exercise training for 15-week. Fecal microbiota composition and function were assessed by 16S rRNA sequencing. The sequencing results showed that SESN2−/− mice displayed differed microbiome profile from WT mice. Exercise enriched the microflora diversity and increased the beneficial microbial species in WT mice, and SESN2 ablation weakened the beneficial effects of exercise on microbial resilience following HFD consumption. Moreover, network analysis revealed that exercise increased correlation density and clustering of operational taxonomic units in WT mice only. KEGG demonstrated that some dominant metabolism-related enzymes and modules increased in SESN2−/− mice. Our results indicated that the effects of exercise on metabolism are associated with the perturbations of gut microbiota composition and function, suggesting that SESN2 contributes to maintain metabolic homeostasis.

Field and laboratory screening of anthurium cultivars for resistance to foliar bacterial blight and the induced activities of defence-related enzymes

Bacterial blight (BB) caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad) is the most destructive disease of ornamental anthurium. In the present study, foliar resistance of 21 anthurium cultivars were assessed under shaded field and laboratory conditions by injection inoculation of 3 × 108 cfu/ml Xad; disease severity was evaluated using a pretransformed rating scale after symptoms survey. Then six selected cultivars with different resistance levels were evaluated for the induced activities of six defence-related enzymes. The obtained results indicated that the same cultivar shared identical resistance under both conditions, but there was a great variation among the cultivars. Anthurium cv. Pink Champion and Manaka showed the highest resistance, and five other cultivars were highly susceptible. The activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) in the resistant cultivars increased much faster and reached much higher peak levels than those in susceptible cultivars. Further analyses revealed that the relative resistance index (RRI) significantly positively correlated with the activities of SOD, APX, POD and PAL, but not with catalase (CAT) and polyphenol oxidase (PPO), suggesting that early rapid accumulation of SOD, APX, PAL and POD might be an important mechanism of defence against Xad and could serve as one of the valuable physiological indices for the prediction of BB resistance in anthurium germplasm. Consequently, the identified resistant cultivars and the induced defence enzymes will facilitate the phytopathological research and enhance blight resistance selection in future breeding

Proteomic Analysis of Skeletal Muscle in Insulin-Resistant Mice: Response to 6-Week Aerobic Exercise

<div><p>Aerobic exercise has beneficial effects on both weight control and skeletal muscle insulin sensitivity through a number of specific signaling proteins. To investigate the targets by which exercise exerts its effects on insulin resistance, an approach of proteomic screen was applied to detect the potential different protein expressions from skeletal muscle of insulin-resistant mice after prolonged aerobic exercise training and their sedentary controls. Eighteen C57BL/6 mice were divided into two groups: 6 mice were fed normal chow (NC) and 12 mice were fed high-fat diet (HFD) for 10 weeks to produce an IR model. The model group was then subdivided into HFD sedentary control (HC, n = 6) and HFD exercise groups (HE, n = 6). Mice in HE group underwent 6 weeks of treadmill running. After 6 weeks, mice were sacrificed and skeletal muscle was dissected. Total protein (n = 6, each group) was extracted and followed by citrate synthase, 2D proteome profile analysis and immunoblot. Fifteen protein spots were altered between the NC and HC groups and 23 protein spots were changed between the HC and HE groups significantly. The results provided an array of changes in protein abundance in exercise-trained skeletal muscle and also provided the basis for a new hypothesis regarding the mechanism of exercise ameliorating insulin resistance.</p> </div

Quadriceps femoris protein profiling by 2-DE.

<p>A typical 2-D-pattern gel image of 100-µg protein extract separated in a pH 3–10 IPG strip in the first dimension and 13% polyacrylamide gel in the second dimension. One 2-D gel was performed each sample, 6 samples per group. Twenty-five differentially expressed (p<0.05) spots were labeled with spot number as they appear in the MS list (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053887#pone-0053887-t002" target="_blank">Table 2</a>).</p

Selected proteins from 2-DE were confirmed by immunoblot analysis.

<p>Expression of Trim72, Myh4, Skeletal Muscle Actin (SM Actin), Hsp25 and Fabp4 were assessed by western blot analysis of skeletal muscle proteins from NC, HC, and HE mice; β-tubulin was used as an internal control for loading.</p

Body weight in the experimental procedure.

<p>Values are means ± SEM (n = 6, each group). *: HC (HFD control) vs. NC (normal chow control) p<0.05; ^#: HE (HFD exercise) vs. HC p<0.05.</p