Carboxylesterase3 (Ces3) Interacts with Bone Morphogenetic Protein 11 and Promotes Differentiation of Osteoblasts via Smad1/5/9 Pathway

술라그나 무케르지,박종필,윤종원 한국생물공학회 2022 Biotechnology and Bioprocess Engineering Vol.27 No.1

Ces3 is a lipolytic enzyme predominantly present in liver and adipocytes, with recent reports of its presence in skeletal muscles, as well. A cross-linking study to understand the various interacting proteins involved in bone-adipose axis could provide novel targets for drug development. We explored the functional role of Ces3 in osteoblasts and mesenchymal stem cells differentiating into osteoblast lineage using in vitro models. We also investigated the physiological functions of Ces3 by stable gene knockdown of Ces3 and exogenous Ces3 induction, and examined the interacting proteins by Co-IP and insilico analysis. Data from our study suggests that Ces3 is highly expressed in osteoblasts and promotes proliferation of the cells by increasing the expressions of osteogenic marker proteins and genes. For the first time, our mechanistic studies revealed that Ces3 interacts with BMP11 protein for regulation of osteoblast differentiation and activates the ALK2 and BMP type II receptors via Smad 1/5/9 signaling pathways. In addition, we identified the various partner proteins linked to Ces3 and BMP11 which are also involved in the metabolic network of osteoblasts. In silico analysis revealed a direct and strong interaction between Ces3 and BMP11 which influences the growth and regulation of osteoblasts. Current data unveiled a hitherto unknown mechanism of Ces3 and BMP11 in the bone-adipose axis, shedding light on Ces3 as a pharmacotherapeutic target to treat metabolic disorders.

Trans-anethole Induces Thermogenesis via Activating SERCA/SLN Axis in C2C12 Muscle Cells

술라그나 무케르지,최민지,윤종원 한국생물공학회 2022 Biotechnology and Bioprocess Engineering Vol.27 No.6

Recently, adaptive non-shivering thermogenesis has attracted considerable attention because it can elevate energy expenditure and help treat obesity. Despite the numerous reports related to UCP1-driven thermogenesis, little is known regarding UCP1-independent thermogenesis in adipose tissues and muscle. Therefore, it is essential to identify the molecular targets for UCP1-independent thermogenesis and their mechanisms to increase the energy expenditure pharmacologically in both adipocytes and muscle. This study examined whether trans-anethole (TA), a major component of the essential oils of fennel, induces UCP1-independent SERCA/SLN-based thermogenesis and promotes the lipid metabolism in muscle cells. TA enhanced myogenesis, lipolysis, and the oxidative metabolism in C2C12 muscle cells. More importantly, TA activated the SERCA/SLN/RYR axis, thereby inducing thermogenesis in muscle cells. Molecular docking analysis revealed a good interaction between SERCA with TA with a strong bind activity. In conclusion, the current data unveiled a previously unknown mechanism of TA in myoblasts and suggests a possible therapeutic agent in muscles by enhancing energy expenditure.

Curcumin Stimulates UCP1-independent Thermogenesis in 3T3-L1 White Adipocytes but Suppresses in C2C12 Muscle Cells

최민지,술라그나 무케르지,윤종원 한국생물공학회 2022 Biotechnology and Bioprocess Engineering Vol.27 No.6

Non-shivering thermogenesis may be an effective way to alter the energy balance in adipocytes and skeletal muscle. Curcumin stimulates adipocyte browning and improves the mitochondrial function in adipocytes via uncoupling protein 1 (UCP1)-dependent thermogenic activity. On the other hand, the UCP1-independent thermogenic effect of curcumin on adipose tissues and muscle remains unexplored. This study examined whether curcumin can also induce UCP1-independent thermogenesis in 3T3-L1 white adipocytes and C2C12 muscle cells. Curcumin stimulated the expression of α1-adrenergic receptor (α1-AR), UCP1-independent thermogenic markers, sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) 2b, and ryanodine receptor 2 in adipocytes, whereas it suppressed SERCA/sarcolipinbased thermogenesis in muscle cells. Curcumin stimulated non-shivering thermogenesis by activating mitochondrial uncoupling and the SERCA/SLN axis in white adipocytes but not muscle cells. In addition, curcumin stimulated thermogenesis by activating the creatine metabolism-related thermogenic pathway in white adipocytes. Taken together, curcumin induces UCP1-independent creatine-mediated and α1-AR/SERCA-based thermogenesis through ATP-consuming futile processes. Together with previous results, the antiobesity effect of curcumin involves mainly two interacting parts, one mediated via β3-AR and cAMP (UCP1-dependent) and the other via α1-AR and increase in cytosolic Ca2+ levels (UCP1-independent) in beige fat but not in muscle cells.

Loss of ADAMTS15 Promotes Browning in 3T3-L1 White Adipocytes via Activation of β3-adrenergic Receptor

최민지,술라그나 무케르지,윤종원 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.2

ADAMTSs belong to the superfamily of secreted metalloendopeptidases, some of which are reported to be closely associated with obesity. However, the role of ADAMTS15 is not well characterized in adipocytes. This study investigates the effect of Adamts15 deficiency on lipid metabolism in 3T3-L1 and HIB1B adipocytes, with a focus on the role of browning of white adipocytes. Quantitative real-time PCR, immunoblot analysis, immunofluorescence, and staining methods were applied to evaluate the effects of ADAMTS15 on other target proteins and genes involved in lipid metabolism, after silencing Adamts15 by applying the siRNA technique. Our results demonstrate that ADAMTS15 is expressed in both white and brown adipocytes, and the deficiency of Adamts15 promotes browning of white adipocytes by enhancing the expression of brown adipocyte-specific genes and proteins. In addition, silencing of Adamts15 activates brown adipocytes and also upregulates lipid metabolic activity in both white and brown adipocytes, by increasing mitochondrial biogenesis as well as the levels of lipolytic and fat oxidative marker proteins, and reducing adipogenic factors. Moreover, mechanistic studies revealed that depletion of Adamts15 induces browning via activation of the β3AR-PKA-CREB signaling pathway. Taken together, our data unveiled a previously unknown mechanism of ADAMTS15 in the regulation of lipids, and the significance of this protein as a pharmacotherapeutic target to treat obesity-related metabolic disorders.

Theobromine, a Methylxanthine in Cocoa Bean, Stimulates Thermogenesis by Inducing White Fat Browning and Activating Brown Adipocytes

장명환,강남현,술라그나 무케르지,윤종원 한국생물공학회 2018 Biotechnology and Bioprocess Engineering Vol.23 No.6

Natural medicinal compounds to treat obesity have recently attracted a great deal of attention because of the serious side effects of synthetic anti-obesity drugs. Recent advances have been made to identify natural products showing thermogenic activity, which is responsible for energy expenditure in brown or brown-like (beige) adipocytes. Here, we explored the thermogenic effects of theobromine, one of the most abundant methylxanthines in cocoa, on 3T3-L1 white adipocytes and HIB1B brown adipocytes. Theobromine markedly increased the expression levels of brown-fat signature proteins (PGC-1α, PRDM16, and UCP1) and beige-specific genes (Cd137, Cidea, Cited1, Tbx1, and Tmen26) in 3T3-L1 white adipocytes and remarkably elevated the expression levels of brown fatspecific genes (Cidea, Lhx8, Ppargc1, Prdm16, Ucp1, and Zic1) in HIB1B brown adipocytes. Theobromine also reduced the expression of the key adipogenic transcription factors, C/EBPα and PPARγ, in white adipocytes, while enhancing their expression in HIB1B cells. In addition, theobromine regulated lipolytic events and fat oxidation by upregulating the expression of pACC, ATGL, pHSL, ACOX, and CPT1. Additional mechanistic study revealed that theobromine activates β3-AR and AMPK. In summary, our results provide evidence for the first time indicating that theobromine has a potential beneficial effect on browning of white adipocytes and improves lipid catabolic metabolism in both cultured white and brown adipocytes via β-adrenergic signaling and AMPK activation. Consumption of theobromine may be a feasible way to activate thermogenesis and improve systematic lipid metabolism to protect against obesity and other metabolic disorders.