HDAC inhibitor Trichostatin A suppresses adipogenesis in 3T3-L1 preadipocytes

Abstract

Background and purpose: Obesity is becoming a major global health issue and is mainly induced by the accumulation of adipose tissues mediated by adipogenesis, which is reported to be regulated by peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT enhancer-binding protein α (C/EBPα). Trichostatin A (TSA) is a novel histone deacetylase inhibitor (HDACI) that was recently reported to exert multiple pharmacological functions. The present study will investigate the inhibitory effect of TSA on adipogenesis, as well as the underlying mechanism. Methods: The adipogenesis of 3T3-L1 cells was induced by stimulation with a differentiation cocktail (DMI) medium for 8 days. MTT assay was used to measure the cell viability and Oil Red O staining was used to evaluate the adipogenesis of 3T3-L1 cells. The total level of triglyceride and released glycerol were detected to evaluate the lipolysis during 3T3-L1 adipogenesis. The expression levels of Leptin , fatty acid-binding protein 4 (FABP4) , and sterol regulatory element-binding protein ( SREBP1C) were determined by qRT-PCR. qRT-PCR assay was utilized to detect the expression levels of PPARγ and C/EBPα in 3T3-L1 cells. A high-fat diet (HFD) was used to construct an obese mice model, followed by the treatment with TSA. HE staining was conducted to evaluate the pathological state of adipose tissues. Body weights and the weights of adipose tissues were recorded to evaluate the anti-obesity property of TSA. Results: Firstly, the promoted lipid accumulation induced by DMI incubation was significantly reversed by the treatment with TSA in a dose-dependent manner. The elevated expression levels of Leptin , FABP4 , SREBP1C, PPARγ, and C/EBPα induced by the stimulation with DMI incubation were dramatically inhibited by the introduction of TSA, accompanied by the upregulation of phosphorylated AMP-activated protein kinase (p-AMPK). Secondly, the inhibitory effect of TSA against the expression level of PPARγ and lipid accumulation was greatly abolished by an AMPK inhibitor. Lastly, the increased body weights and visceral adipocyte tissue weight, as well as the enlarged size of adipocytes induced by HFD were pronouncedly reversed by the administration of TSA. Conclusion: TSA inhibited adipogenesis in 3T3-L1 preadipocytes by activating the AMPK pathway.

Trichostatin D as a Novel KLF2 Activator Attenuates TNFα-Induced Endothelial Inflammation

“…Histone deacetylases catalyze DNA deacetylation and play a crucial role in the modulation of atherosclerosis [ 31 ]. TSD is an analog of trichostatin A (TSA), which is a pan-HDAC inhibitor [ 32 , 33 , 34 ]. Studies have found that inhibition of HDAC activity induced KLF2 expression, which then contributed to reducing inflammation of endothelial cells [ 35 , 36 ].…”

“…TSD is a structural analog of TSA. TSA is a known pan-inhibitor of HDACs that thereby regulating the expression of related genes [ 32 , 33 , 34 ]. It was reported that HDAC4, 5, and 7 inhibited KLF2 expression, which in turn had an important role in anti-inflammation, sepsis, and angiogenesis [ 16 , 35 , 42 , 43 , 44 ].…”

Reducing White Adipose Tissue Browning Using p38α MAPK Inhibitors Ameliorates Cancer-Associated Cachexia as Assessed by Magnetic Resonance Imaging

“…iWAT was divided into smaller pieces and digested in phosphate-buffered saline (PBS, Hyclone, Logan, UT, USA) with 0.5 mg/mL collagenase I (Worthington Biochemical Corporation, Lakewood, NJ, USA) at 37 °C for 1.5 h. After digestion, samples were filtered sequentially through nylon filters with a pore size of 600 and 50 µm, and centrifuged at 800 rpm for 5 min. According to the typical induction cocktail method [ 20 , 21 , 22 ], the obtained SVF cell pellet was then resuspended in the complete medium, which containing 89% Dulbecco’s modified Eagle’s medium (DMEM/F12, Gibco, Grand Island, NY, USA), 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA) and 1% penicillin and streptomycin (PS, Hyclone, Logan, UT, USA). Resuspended SVF cells were incubated in 6-well plates at a concentration of 8 × 10 4 cells/well maintained at 37 °C in the presence of 5% CO 2 .…”

Epigenetic regulation in metabolic diseases: mechanisms and advances in clinical study

“…400 Trichostatin A (TSA) is a novel HDACi and reportedly inhibits adipogenesis in 3T3-L1 preadipocytes through the AMPK signalling pathway. 401 By inhibiting HDAC and increasing insulin sensitivity, sodium acetate alleviates hepatic lipotoxicity related to diabetes. 402 Puerarin may attenuate diabetic osteoporosis by preventing inflammation and apoptosis through HDAC1 and HDAC3 inhibition.…”