[논문]Myonectin inhibits adipogenesis in 3T3-L1 preadipocytes by regulating p38 MAPK pathway

Park, Tae-Jun; Park, Anna; Kim, Jaehoon; Kim, Jeong-Yoon; Han, Baek Soo; Oh, Kyoung-Jin; Lee, Eun Woo; Lee, Sang Chul; Bae, Kwang-Hee; Kim, Won Kon

doi:10.5483/bmbrep.2021.54.2.262

[국내논문] Myonectin inhibits adipogenesis in 3T3-L1 preadipocytes by regulating p38 MAPK pathway 원문보기

BMB reports, v.54 no.2, 2021년, pp.124 - 129

Park, Tae-Jun (Department of Microbiology and Molecular Biology, Chungnam National University) , Park, Anna (Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST)) , Kim, Jaehoon (Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST)) , Kim, Jeong-Yoon (Department of Microbiology and Molecular Biology, Chungnam National University) , Han, Baek Soo (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Oh, Kyoung-Jin (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Lee, Eun Woo (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Lee, Sang Chul (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Bae, Kwang-Hee (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Kim, Won Kon (Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))

Abstract ▼ AI-Helper

In current times, obesity is a major health problem closely associated with metabolic disease such as diabetes, dyslipidemia, and cardiovascular disease. The direct cause of obesity is known as an abnormal increase in fat cell size and the adipocyte pool. Hyperplasia, the increase in number of adipocytes, results from adipogenesis in which preadipocytes differentiate into mature adipocytes. Adipogenesis is regulated by local and systemic cues that alter transduction pathways and subsequent control of adipogenic transcription factors. Therefore, the regulation of adipogenesis is an important target for preventing obesity. Myonectin, a member of the CTRP family, is a type of myokine released by skeletal muscle cells. Although several studies have shown that myonectin is associated with lipid metabolism, the role of myonectin during adipogenesis is not known. Here, we demonstrate the role of myonectin during adipocyte differentiation of 3T3-L1 cells. We found that myonectin inhibits the adipogenesis of 3T3-L1 preadipocytes with a reduction in the expression of adipogenic transcription factors such as C/EBPα, β and PPARγ. Furthermore, we show that myonectin has an inhibitory effect on adipogenesis through the regulation of the p38 MAPK pathway and CHOP. These findings suggest that myonectin may be a novel therapeutic target for the prevention of obesity.

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표/그림 (4)

그림 Fig. 1. Suppression of adipogenic differentiation by treatment with myonectin. (A) Effects of myonectin on cell viability. The viability of untreated control cells (Non) was defined as 100%. (B) Effects of myonectin on the differentiation of 3T3-L1 cells. Differentiation of confluent 3T3-L1 cells was induced by the MDI cocktail (0.5 mM IBMX, 1 μM Dexamethasone, and 10 μg/ml insulin). The 3T3-L1 cells were treated with different concentrations (0.05-2 μg/ml) of myonectin during differentiation. On day 10 of differentiation, the fully differentiated adipocytes were fixed and stained with oil red O solution for visualization of the lipid accumulation. (C-F) mRNA levels of adipogenic genes in mature 3T3-L1 adipocytes. Each gene was normalized to Rpl32. All quantitative data are the means ± SD (n = 6). *P < 0.05, **P < 0.005, ***P < 0.0005 compared to untreated control cells on day 0 (Day 0, Non).
그림 Fig. 2. Inhibitory function of myonectin involved in the early stage of adipogenesis. (A) Scheme for the preadipocyte differentiation and myonectin treatment. (B) Oil red O staining of lipid droplets after differentiation of the 3T3-L1 adipocytes treated with and without myonectin. (C) The levels of lipid accumulation of the 3T3-L1 mature adipocytes were quantified at 490 nm. (D) Protein levels and (E-G) mRNA levels of the adipogenic genes in mature 3T3-L1 adipocytes treated with and without myonectin. Each gene was normalized to Rpl32. All quantitative data are the means ± SD (n = 3-6). *P < 0.05, **P < 0.005, ***P < 0.0005 compared to the untreated control cells (Non) on day 10.
그림 Fig. 3. Effect of myonectin on several pathways related to adipogenesis. (A) 3T3-L1 preadipocytes were induced to differentiate by the MDI cocktail with or without myonectin (1 μg/ml). Western blot analyses of p-AKT, p-AMPK and β-Catenin during the initial time of differentiation in the absence or presence of myonectin. Total AKT, total AMPK and HSP90 were presented as the controls for the normalization. (B-D) The ratios of p-AKT to total AKT, p-AMPK to total AMPK, and β-Catenin to HSP90. All quantitative data are the means ± SD (n = 3). *P < 0.05 compared to the myonectin-untreated control cells (MDI).
그림 Fig. 4. Inhibitory function of myonectin on adipogenesis through the p38 MAPK pathway. (A) 3T3-L1 preadipocytes were induced to differentiate by the MDI cocktail with or without myonectin (1 μg/ ml). Western blot analyses of p-p38, p-ERK, p-JNK and CHOP during the initial time of differentiation in the absence or presence of myonectin. Total p38, total ERK and total JNK and HSP90 were presented as controls for normalization. (B, C) The ratios of p-p38 to total p38 and CHOP to HSP90. (D, E) mRNA levels and (F) protein levels of C/EBPβ and PPAR𝛾. Each mRNA expression was normalized to Rpl32. (G, H) The ratios of C/EBPβ and PPAR𝛾 to HSP90. All quantitative data are the means ± SD (n = 3). *P < 0.05, **P < 0.005 compared to the myonectin-untreated control cells (MDI, 0 min).

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문제 정의

Another study proposed that the plasma myonectin concentration could be a major parameter associated with metabolic syndrome and insulin resistance (4). Based on the relationship between myonectin and lipid metabolism, this study aimed to investigate a potential role for myonectin during adipogenesis.

가설 설정

The viability of untreated control cells (Non) was defined as 100%. (B) Effects of myonectin on the differentiation of 3T3-L1 cells. Differentiation of confluent 3T3-L1 cells was induced by the MDI cocktail (0.

제안 방법

To investigate the effect of myonectin on 3T3-L1 cell viability, cytotoxicity was measured by quantitating the amount of ATP present in cells, which indicates the presence of metabolically active cells. When the proliferation rate of the control cells not treated with myonectin was considered to be 100%, the cells exhibited a cell viability of more than 95% at all different concentrations of myonectin (0.
Adiponectin is an adipokine secreted from mature adipocytes such as Leptin and Resistin and is a marker of adipocyte terminal differentiation (13). To further investigate the inhibitory effect of preadipocyte differentiation by myonectin, the expression of adipogenic transcription factors and related genes was measured at different time points (day 0, 2, 6, and 10) during dif- ferentiation. As a result, the mRNA expression levels of PPAR, C/EBP, FABP4, and Adiponectin in the untreated cells were increased as differentiation progressed, but the cells treated with myonectin showed a decrease at all time points (Fig.

데이터처리

The statistical significance of the comparisons was determined using the Student’s two-tailed t-test. A P-value less than 0.

후속연구

by p38. Therefore, further studies are required for deeper understanding of the mechanism. Nevertheless, our findings have open the possibility of myonectin being a potential novel therapeutic target to prevent obesity by showing a distinct inhibitory effect on the differentiation of 3T3-L1 cells.

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