Location: Healthy Body Weight ResearchTitle: Gene and protein profiling of the effects of tart cherry anthocyanins in on preadipocytes Author
|Jayarathne, Shasika - Texas Tech University|
|Scoggin, Shane - Texas Tech University|
|Zabet, Masoud - Texas Tech University|
|Bhawel, Ruchika - Texas Tech University|
|Moustaid-moussa, Naima - Texas Tech University|
Submitted to: Journal of Federation of American Societies for Experimental Biology
Publication Type: Abstract Only
Publication Acceptance Date: 12/1/2016
Publication Date: 4/1/2017
Citation: Jayarathne, S., Scoggin, S., Zabet, M., Bhawel, R., Larson, K.J., Moustaid-Moussa, N. 2017. Gene and protein profiling of the effects of tart cherry anthocyanins in on preadipocytes [abstract]. Journal of Federation of American Societies for Experimental Biology. 31:625.4.
Technical Abstract: Several dietary bioactive compounds possess anti-inflammatory and anti-obesity properties and could potentially reduce obesity-associated cardiovascular diseases, diabetes and other metabolic inflammatory diseases. We are specifically interested in tart cherry (TC) anthocyanins (ACY) and in understanding mechanisms mediating their beneficial effects in metabolic diseases. Hence, we hypothesized that tart cherry ACY reduce obesity-associated inflammation through direct anti-inflammatory and lipid metabolizing effects in adipocytes. We first analyzed major ACY in TC by High Performance Liquid Chromatography (HPLC) and identified cyanidin-3-O-glucoside (C3G), cyanidin-3-O-rutinoside (C3R), cyanidin as well as the polyphenol p-Coumaric acid with C3G being the most abundant ACY in TC. To further gain insight into global effects of tart cherry ACY on adipocyte metabolism, we conducted RNA Seq (using Illumina Hi-Seq platform) and Proteomics (using Liquid chromatography mass spectrometer, LC-MS). We used 3T3-L1 preadipocytes treated without or with 200ng/ml lipopolysaccharide (LPS) for 4hrs to stimulate inflammation, then treated them for 18hrs with TC extracts containing ~18ug/ml ACY. Media were used to determine changes in secreted adipokines by ELISA. TC significantly reduced LPS-induced secretion of the inflammatory cytokine (Interleukin-6, IL-6). Whole genome mRNA sequencing identified several genes differentially regulated by tart cherry ACY. Among 36,919 expressed mus-musculus genes, 2359 genes were identified as differentially expressed at the fold-change of 2 and 95% confidence level using RNA sequencing. Of these, 14 genes were up regulated and 2345 genes were down regulated by tart cherry ACY. Using Ingenuity Pathway Analysis (IPA®), we identified predicted inhibition of inflammatory, immune and oxidative stress-related pathways and genes by TC (e.g. Protein Kinase,TNFR2 and B cell activating factor signaling, genes: casp14, 14-3-3, c-Fos, c-Jun). Using proteomics, we identified 62 differentially regulated proteins. Among these, inflammatory, oxidative stress and apoptotic related proteins were down-regulated at p<0.05 (e.g. NRF2, GSTP1, ASNS, HSP90a etc.). In conclusion, our results demonstrate that tart cherry ACY reduce expression of genes, proteins and pathways associated with inflammation and oxidative stress. Gene and protein profiling approaches help identify novel pathways that may potentially serve as new targets for obesity and inflammation.