Trapa japonica Flerov Extract Prevents Obesity by Regulating Adipogenesis and Lipolysis in Differentiated 3T3-L1 Cells

Our study investigated that the anti-obesity effect of the Trapa japonica Flerov extract (TJ) in differentiated 3T3-L1 adipocytes. To this end, 3T3-L1 cells were treated with TJ during their differentiation period. On the last day of the cell culture, we tested intracellular cAMP, FA, glycerol release, TG, and performed Oil Red O staining and Western blot assays. On the part of adipogenesis, lipogenesis, and lipolysis mechanism, TJ increased the cAMP (maximum 125.4%) levels and glycerol release (maximum four times) and decreased FA (maximum 35.1%) and TG (maximum 35.7%) levels. Furthermore, the protein expression levels of each mechanism-related factor were regulated in a dose-dependent manner. These results indicate that TJ reduced lipid accumulation by max 53.6% and 47.9%, respectively, in adipogenesis and lipolysis mechanisms. We expect this effect of TJ to be due to its component, ellagic acid. In conclusion, we found that TJ inhibits TG synthesis during adipogenesis and lipogenesis, promotes lipolysis, and thus, indicating its potential as a functional food for obesity prevention.

Degradation of ribosome and chaperone proteins is attenuated during the Differentiation of Replicatively Aged C2C12 Myoblasts

Age-related impairments in myoblast differentiation may contribute to reductions in muscle function in older adults, however, the underlying proteostasis processes are not well understood. Young (P6-10) and replicatively aged (P48-50) C2C12 myoblast cultures were investigated during early (0h-24h) and late (72h-96h) stages of differentiation using deuterium oxide (D2O) labelling and mass spectrometry. The absolute dynamic profiling technique for proteomics (Proteo-ADPT) was applied to quantify the absolute rates of abundance change, synthesis and degradation of individual proteins. Proteo-ADPT encompassed 116 proteins and 74 proteins exhibited significantly (P<0.05, FDR <5 %) different changes in abundance between young and aged cells at early and later periods of differentiation. Young cells exhibited a steady pattern of growth, protein accretion and fusion, whereas aged cells failed to gain protein mass or undergo fusion during later differentiation. Maturation of the proteome was retarded in aged myoblasts at the onset of differentiation, but their proteome appeared to "catch up" with the young cells during the early phase of the differentiation period. However, this "catch up" process in aged cells was not accomplished by higher levels of protein synthesis. Instead, a lower level of protein degradation in aged cells was responsible for the elevated gains in protein abundance. Our novel data point to a loss of proteome quality as a precursor to the lack of fusion of aged myoblasts and highlights dysregulation of protein degradation, particularly of ribosomal and chaperone proteins, as a key mechanism that may contribute to age-related declines in the capacity of myoblasts to undergo differentiation.

Irisin Stimulates the Release of CXCL1 From Differentiating Human Subcutaneous and Deep-Neck Derived Adipocytes via Upregulation of NFκB Pathway

Thermogenic brown and beige adipocytes might open up new strategies in combating obesity. Recent studies in rodents and humans have indicated that these adipocytes release cytokines, termed “batokines”. Irisin was discovered as a polypeptide regulator of beige adipocytes released by myocytes, primarily during exercise. We performed global RNA sequencing on adipocytes derived from human subcutaneous and deep-neck precursors, which were differentiated in the presence or absence of irisin. Irisin did not exert an effect on the expression of characteristic thermogenic genes, while upregulated genes belonging to various cytokine signaling pathways. Out of the several upregulated cytokines, CXCL1, the highest upregulated, was released throughout the entire differentiation period, and predominantly by differentiated adipocytes. Deep-neck area tissue biopsies also showed a significant release of CXCL1 during 24 h irisin treatment. Gene expression data indicated upregulation of the NFκB pathway upon irisin treatment, which was validated by an increase of p50 and decrease of IκBα protein level, respectively. Continuous blocking of the NFκB pathway, using a cell permeable inhibitor of NFκB nuclear translocation, significantly reduced CXCL1 release. The released CXCL1 exerted a positive effect on the adhesion of endothelial cells. Together, our findings demonstrate that irisin stimulates the release of a novel adipokine, CXCL1, via upregulation of NFκB pathway in neck area derived adipocytes, which might play an important role in improving tissue vascularization.

Effect of Sake Lees on the Inhibition of Lipid Accumulation in Adipocytes

Recent lifestyle changes, such as the Westernization of diets and the rise in the prevalence of obesity, with an associated increase in the number of patients with lifestyle-related diseases, have become a social issue. Fermented food has attracted attention as a solution to problems caused by obesity. Sake lees, a byproduct of sake brewing, represent one such food that is expected to have health benefits. In this study, we investigated the effects of sake lees components on preadipocytes (3T3-L1). We cultured preadipocytes in a medium with indigestible sake lees components (ISLCs) to investigate lipid accumulation, analyzed the glycerol 3-phosphate dehydrogenase (GPDH) and LPL activities of those cells, and performed a real-time PCR analysis of the IL6 expression in the cells. The results show that lipid accumulation and GPDH activity were significantly decreased in adipocytes treated with 1.0 mg/mL ISLCs compared to untreated cells. Furthermore, the expression of IL6 in adipocytes treated with 1.0 mg/mL ISLCs was significantly decreased and the lipase activity was significantly increased in adipocytes treated with ISLCs after differentiation. IL6 is known to have multiple functions in adipose tissue. In conclusion, ISLCs were associated with reduced lipid accumulation in adipocytes, with effects on IL6 expression and LPL activity observed throughout the differentiation period.

Genome-wide characterization and expression profiling of B3 superfamily during ethylene-induced flowering in pineapple (Ananas comosus L.)

Background The B3 superfamily (B3s) represents a class of large plant-specific transcription factors, which play diverse roles in plant growth and development process including flowering induction. However, identification and functional surveys of B3 superfamily have not been reported in ethylene-induced pineapple flowering (Ananas comosus). Results 57 B3 genes containing B3 domain were identified and phylogenetically classified into five subfamilies. Chromosomal localization analysis revealed that 54 of 57 AcB3s were located on 21 Linkage Groups (LG). Collinearity analysis demonstrated that the segmental duplication was the main event in the evolution of B3 gene superfamily, and most of them were under purifying selection. The analysis of cis-element composition suggested that most of these genes may have function in response to abscisic acid, ethylene, MeJA, light, and abiotic stress. qRT-PCR analysis of 40 AcB3s containing ethylene responsive elements exhibited that the expression levels of 35 genes were up-regulated within 1 d after ethephon treatment and some were highly expressed in flower bud differentiation period in stem apex, such as Aco012003, Aco019552 and Aco014401. Conclusion This study provides a basic information of AcB3s and clues for involvement of some AcB3s in ethylene-induced flowering in pineapple.

Irisin stimulates the release of CXCL1 from differentiating human subcutaneous and deep-neck derived adipocytes via upregulation of NFκB pathway

Thermogenic brown and beige adipocytes play an important role in combating obesity. Recent studies in rodents and humans have indicated that these adipocytes release cytokines, termed "batokines". Irisin was discovered as a polypeptide regulator of beige adipocytes released by myocytes, primarily during exercise. We performed global RNA sequencing on adipocytes derived from human subcutaneous and deep-neck precursors, which were differentiated in the presence or absence of irisin. Irisin did not exert an effect on the expression of characteristic thermogenic genes, while upregulated genes belonging to various cytokine signaling pathways. Out of the several upregulated cytokines, CXCL1, the highest upregulated, was released throughout the entire differentiation period, and predominantly by differentiated adipocytes. Deep-neck area tissue biopsies also showed a significant release of CXCL1 during 24 hours irisin treatment. Gene expression data indicated upregulation of the NFκB pathway upon irisin treatment, which was validated by an increase of p50 and decrease of IκBα protein level, respectively. Continuous blocking of the NFκB pathway, using a cell permeable inhibitor of NFκB nuclear translocation, significantly reduced CXCL1 release. The released CXCL1 exerted a positive effect on the adhesion of endothelial cells. Together, our findings demonstrate that irisin stimulates the release of a novel "batokine", CXCL1, via upregulation of NFκB pathway in neck area derived adipocytes, which might play an important role in improving tissue vascularization.

Indian Gooseberry and Barley Sprout Mixture Inhibits Adipogenesis and Lipogenesis Activity in 3T3-L1 Adipocytes

This study aimed to confirm the synergistic effect of an Indian gooseberry (IG) and barley sprout (BP) mixture in differentiated adipocytes. To this end, 3T3-L1 adipocytes were treated with IG, BP, and IGBP mixtures during the differentiation period. On the last day of differentiation, we measured intracellular cAMP, triglyceride (TG), and fatty acid (FA) levels, as well as performed Oil Red O staining, glycerol release, and Western blot assays. During adipogenesis, IGBP (200 μg/mL) increased the cAMP levels by more than 2-fold and decreased the protein expressions levels of p-CREB (66.3%), C/EBPα (79.4%), C/EBPβ (85.9%), and PPARγ (74.1%) compared to those in the C group. Furthermore, the expression levels of the adipogenesis-related genes and GLUT4 (more than 3-fold) were regulated. During lipogenesis, the IGBP (200 μg/mL) activated AMPK and ACC levels and reduced the protein expression levels of SREBP1c, FAS, and LPL. This reduced the FA and TG contents in the cells by 47.6% and 76.3%, respectively, compared to those in the differentiated control (C) group, resulting in a more than 5-fold increase in glycerol release. In conclusion, we found that IGBP inhibited TG synthesis during adipogenesis and lipogenesis, and thus, displayed potential as a functional health food for preventing obesity.

Genome-wide characterization and expression profiling of B3 superfamily during flower induction stage in pineapple (Ananas comosus L.)

The B3 superfamily is a plant-specific family, which involves in growth and development process. By now, the identification of B3 superfamily in pineapple (Ananas comosus) has not reported. In this study, 57 B3 genes were identified and further phylogenetically classified into five subfamilies, all genes contained B3 domain. Chromosomal localization analysis revealed that 54 of 57 AcB3 genes were located on 21 chromosomes.Collinearity analysis indicated that the segmental duplication was the main event in the evolution of B3 gene superfamily and most of them were under purifying selection. Moreover, there were 7 and 39 pairs of orthologous B3s were identified between pineapple and Arabidopsis or rice, respectively, which indicated the closer genetic relationship between pineapple and rice. Most genes had cis-element of abscisic acid, ethylene, MeJA, light, and abiotic stress. qRT-PCR showed that the expression level of most AcB3 genes were up-regulated within 1 d after ethephon treatment and expressed high level in flower bud differentiation period in stem apex. This study provide a comprehensive understanding of AcB3s and a basis for future molecular studies of ethephon induced pineapple flowering.