Mulberry 1-Deoxynojirimycin Inhibits Adipogenesis by Repression of the ERK/PPARγ Signaling Pathway in Porcine Intramuscular Adipocytes

Wang, Guoqiang; Zhu, Li; Ma, Ming; Chen, Xiaochang; Gao, Yun; Yu, Taiyong; Yang, Gongshe; Pang, Wei

doi:10.1021/acs.jafc.5b01680

Cited by 52 publications

(31 citation statements)

References 43 publications

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“…Cells were resuspended in DMEM/F12 and plated at a density 6 × 10 5 per 60-mm culture dish, and cultured in a 5% CO 2 incubator at 37 °C. Because preadipocytes attach much earlier than myoblasts, the cultures cells were rinsed with PBS three times 1 h after plating to remove insoluble myofibrillar proteins and other insoluble debris [ 2 , 22 , 37 ]. Cells were cultured in growth medium until they reached 80% confluence and digested with 0.05% trypsin, which contained 0.5 mmol/L EDTA, collected by centrifugation 1000× g for 5 min, then resuspended in growth medium and plated at a density of 5 × 10 4 cells/cm in 6-well plate and used to induce differentiation for research.…”

Section: Methodsmentioning

confidence: 99%

miR-425-5p Inhibits Differentiation and Proliferation in Porcine Intramuscular Preadipocytes

Chen

Xiong

Peng

et al. 2017

IJMS

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Intramuscular fat (IMF) content affects the tenderness, juiciness, and flavor of pork. An increasing number of studies are focusing on the functions of microRNAs (miRs) during porcine intramuscular preadipocyte development. Previous studies have proved that miR-425-5p was enriched in porcine skeletal muscles and played important roles in multiple physiological processes; however, its functions during intramuscular adipogenesis remain unclear. To explore the role of miR-425-5p in porcine intramuscular adipogenesis, miR-425-5p agomir and inhibitor were used to perform miR-425-5p overexpression and knockdown in intramuscular preadipocytes, respectively. Our results showed that the agomir of miR-425-5p dramatically inhibited intramuscular adipogenic differentiation and downregulated the expression levels of adipogenic marker genes PPARγ, FABP4, and FASN, whereas its inhibitor promoted adipogenesis. Interestingly, the agomir repressed proliferation of porcine intramuscular preadipocytes by downregulation of cyclin B and cyclin E. Furthermore, we demonstrated that miR-425-5p inhibited adipogenesis via targeting and repressing the translation of KLF13. Taken together, our findings identified that miR-425-5p is a novel inhibitor of porcine intramuscular adipogenesis possibly through targeting KLF13 and subsequently downregulating PPARγ.

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Section: Methodsmentioning

confidence: 99%

miR-425-5p Inhibits Differentiation and Proliferation in Porcine Intramuscular Preadipocytes

Chen

Xiong

Peng

et al. 2017

IJMS

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“…For Frozen section and HE staining of mouse Quad, Sol, Gast, Ing sections were collected on day 180 for Sirt1 AS lncRNA overexpression. Frozen section and HE staining were performed according to the recently published method 59 . The sections were observed and taken through pictures with microscope (Olympus, New York, NY).…”

Section: Methodsmentioning

confidence: 99%

Sirt1 AS lncRNA interacts with its mRNA to inhibit muscle formation by attenuating function of miR-34a

Wang

Xiong

et al. 2016

Sci Rep

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107

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Recent studies demonstrate the functions of long non-coding RNAs (lncRNAs) in mediating gene expression at the transcriptional or translational level. Our previous study identified a Sirt1 antisense (AS) lncRNA transcribed from the Sirt1 AS strand. However, its role and regulatory mechanism is still unknown in myogenesis. Here, functional analyses showed that Sirt1 AS lncRNA overexpression promoted myoblast proliferation, but inhibited differentiation. Mechanistically, Sirt1 AS lncRNA was found to activate its sense gene, Sirt1. The luciferase assay provided evidences that Sirt1 AS lncRNA interacted with Sirt1 3′ UTR and rescued Sirt1 transcriptional suppression by competing with miR-34a. In addition, RNA stability assay showed that Sirt1 AS lncRNA prolonged Sirt1 mRNA half-life from 2 to 10 h. Ribonuclease protection assay further indicated that it fully bound to Sirt1 mRNA in the myoblast cytoplasm. Moreover, Sirt1 AS overexpression led to less mouse weight than the control because of less lean mass and greater levels of Sirt1, whereas the fat mass and levels of miR-34a were not altered. Based on the findings, a novel regulatory mechanism was found that Sirt1 AS lncRNA preferably interacted with Sirt1 mRNA forming RNA duplex to promote Sirt1 translation by competing with miR-34a, inhibiting muscle formation.

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“…Porcine intramuscular adipocyte were isolated from the LD muscle of HN pigs at 2 days of age following methods as previously described [23,24]. Intramuscular adipocytes were cultured with the normal medium: DMEM/F12 supplemented with FBS (10%) and antibiotics (penicillin, 100 IU/mL; streptomycin, 100 µg/mL) at 37 °C and 5% CO 2 in a normal-atmosphere incubator.…”

Section: Isolation Differentiation and Culture Of Porcine Intramuscmentioning

confidence: 99%

LncRNA IMFlnc1 promotes porcine intramuscular adipocyte differentiation by sponging miR-199a-5p to up-regulate CAV-1

Wang

Chen

et al. 2020

Preprint

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Background Local Chinese local pig breeds have better meat quality, such as thinner muscle fiber and higher intramuscular-fat (IMF) content. However, its molecular regulation mechanism has not been discussed in-depth. Studies indicated that long non coding RNAs (lncRNAs) participate in the regulation of muscle and fat development. The expressional differences of lncRNAs in the longissimus dorsi (LD) muscle were identified between Huainan pigs (local Chinese pigs, fat-type, HN) and Large White pigs (lean-type, LW) at 38, 58, and 78 days post conception (dpc).Results In total, 2131 novel lncRNAs were identified in 18 samples, and 291, 305, and 683 differentially expressed lncRNAs (DELs) were found between these two breeds at three stages, respectively. GO and KEGG analysis of the DEL co expressed mRNAs showed that muscle development and energy metabolism were more active at 58 dpc in HN, but at 78 dpc in LW pigs. Muscle cell differentiation and myofibril assembly might contribute to earlier myogenesis and primary-muscle-fiber assembly in HN, and cell proliferation, insulin, and the mitogen-activated protein kinase (MAPK) pathway might contribute to longer proliferation in LW pigs. The PI3K/Akt and cAMP pathways were associated with higher IMF deposition in HN. IMFlnc1 was selected for functional verification, and results indicated that it regulated the expressional level of caveolin-1 (CAV-1) as a competing endogenous RNA (ceRNA) for miR-199a-5p.Conclusion Our data contributed to understanding lncRNAs in porcine-muscle development and IMF deposition, and provided valuable information for improving pig-meat quality.

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Mulberry 1-Deoxynojirimycin Inhibits Adipogenesis by Repression of the ERK/PPARγ Signaling Pathway in Porcine Intramuscular Adipocytes

Cited by 52 publications

References 43 publications

miR-425-5p Inhibits Differentiation and Proliferation in Porcine Intramuscular Preadipocytes

miR-425-5p Inhibits Differentiation and Proliferation in Porcine Intramuscular Preadipocytes

Sirt1 AS lncRNA interacts with its mRNA to inhibit muscle formation by attenuating function of miR-34a

LncRNA IMFlnc1 promotes porcine intramuscular adipocyte differentiation by sponging miR-199a-5p to up-regulate CAV-1

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