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Effect of Blueberry Polyphenols on 3T3-F442A Preadipocyte Differentiation
Shiwani S. Moghe,Shanil Juma,Victorine Imrhan,Parakat Vijayagopal 한국식품영양과학회 2012 Journal of medicinal food Vol.15 No.5
Today obesity is an epidemic, and its prevalence has increased significantly over the last few decades. To avoid excessive accumulation of fat, optimum energy intake along with regular exercise is mandatory. Polyphenols present in green tea, grape seeds, orange, and grapefruit combat adipogenesis at the molecular level and also induce lipolysis. However,very little is known regarding the role of blueberry polyphenols on adipocyte differentiation. Hence we tested the dosedependent effects of blueberry polyphenols on mouse 3T3-F442A preadipocyte differentiation and lipolysis. 3T3-F442A preadipocytes were incubated with three doses of blueberry polyphenols (150, 200, and 250 lg/mL [BB-150, BB-200, and BB-250, respectively]), and intracellular lipid content, cell proliferation, and lipolysis were assayed. Blueberry polyphenols suppressed adipocyte differentiation determined by Oil Red-O staining and AdipoRed assay. Intracellular lipid content in control (11,385.51 – 1,169.6 relative fluorescence units) was significantly higher (P < .05) than with the three doses of blueberry polyphenols (8336.86 – 503.57, 4235.67 – 323.17, and 3027.97 – 346.61, respectively). This corresponds to a reduction of 27%, 63%, and 74%, respectively. Cell proliferation was observed to be significantly higher in the control (0.744 – 0.035 optical density units) than with BB-150 (0.517 – 0.031), BB-200 (0.491 – 0.023), and BB-250 (0.455 – 0.012). However, when tested for lipolysis, there was no significant difference observed among the groups. We conclude that blueberry polyphenols may play an effective role in inhibiting adipogenesis and cell proliferation.
Supriya Shiwani,Naresh Kumar Singh,Myeong Hyeon Wang 한국영양학회 2012 Nutrition Research and Practice Vol.6 No.5
The study elucidated carbohydrase inhibition, anti-cancerous, free radical scavenging properties and also investigated the DNA and protein protection abilities of methanolic root extract of Rumex crispus (RERC). For this purpose, pulverized roots of Rumex crispus was extracted in methanol (80% and absolute conc.) for 3 hrs for 60℃ and filtered and evaporated with vacuum rotary evaporator. RERC showed high phenolic content (211 ㎍/GAE equivalent) and strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50 = 42.86 (absolute methanol) and 36.91 ㎍/mL (80% methanolic extract)) and reduced power ability. Furthermore, RERC exhibited significant protective ability in H₂O₂/Fe<SUP>3+</SUP>/ascorbic acid-induced protein or DNA damage and percentage inhibition of the HT-29 cell growth rate following 80% methanolic RERC exposure at 400 ㎍/mL was observed to be highest (10.2%± 1.03). Moreover, methanolic RERC inhibited α-glucosidase and amylase effectively and significantly (P < 0.05). Conclusively, RERC could be considered as potent carbohydrase inhibitor, anti-cancerous and anti-oxidant.
Reza, A. M.,Shiwani, S.,Singh, N. K.,Lohakare, J. D.,Lee, S. J.,Jeong, D. K.,Han, J. Y.,Rengaraj, D.,Lee, B. W. Springer Science + Business Media 2014 In vitro cellular & developmental biology Animal Vol.50 No.3
The study was conducted to know and investigate the mechanism involved during mesenchymal to epithelial transition to unravel questions related to mammary gland development in prepubertal Korean black goat. We, therefore, biopsied mammary fat pad and isolated adipose cells and characterized with stemness factors (CD34, CD13, CD44, CD106, and vimentin) immunologically and through their genetic expression. Furthermore, characterized cells were differentiated to adipogenic (thiazolidinediones and alpha-linolenic acid) and epithelial (keratinocyte growth factor) lineages. Thiazolidinediones/or in combination with alpha-linolenic acid demonstrated significant upregulation of adipo-Q, PPAR-gamma, CEBP-alpha, LPL, and resistin. Adipose stem cells in induction mixture (5 mu g/ml insulin, 1 mu g/ml hydrocortisone, and 10 ng/ml epidermal growth factor) and subsequent treatment with 10 ng/ml keratinocyte growth factor revealed their trans-differentiating ability to epithelial lineage. From 2 d onwards, the cells under keratinocyte growth factor influenced cells to assume rectangular (2-4 d) to cuboidal (8-10 d) shapes. Ayoub-Shklar stain developed brownish-red pigment in the transformed cells. Though, expressions of K8 and K18 were noted to be highly significant (p < 0.01) but expressions of epithelial membrane antigens and epithelial specific antigens were also significant (p < 0.05) compared to 0 d. Conclusively, epithelial transformations of mammary adipose stem cells would add up knowledge to develop therapeutic regimen to deal with mammary tissue injury and diseases.
Singh, N K,Shiwani, S,Hwang, I H Springer 2012 In vitro cellular & developmental biology Animal Vol.48 No.3
<P>Studies on skeletal muscle cell specification and development have demonstrated in the past that calpains interact with various transcriptional factors in regulating the cellular function. It has therefore, been assumed that transcriptional factors like myogenin, MyoD, Myf5, and MRF4 that are active during the myogenic differentiation might be affected and degraded by calpains. Therefore, to examine the biochemical adaptations of myoblasts during myocyte formation and muscle development comprehensively, the current study was designed to identify the effect of calpeptin (calpain inhibitors) on protein expression during differentiation of C2C12 mouse myoblast. Cells were proliferated to near 80% confluence under Dulbecco's modified eagle medium and differentiated further in 2% HS with 50?μM calpeptin. Incubated cells were collected at 0, 12, and 72?h and later the cell proteins were focused onto pH?4-7 IEF strip, followed by 12.5% SDS-PAGE. Obtained spots on the gels were compared and matched using commercial 2-DE analysis software and matched spots were identified by MALDI-ToF and/or Q-Tof systems. Conclusively, cell differentiation was observed to be active from 12 to 72?h however, calpeptin affected the differentiation process and cut down the rate of fusion by approximately 50%. Out of 41 proteins identified, 12 proteins were found to be upregulated where as 29 proteins were downregulated.</P>
Transdifferentiation of bovine epithelial cells towards adipocytes in the presence of myoepithelium
Sugathan, Subi,Lee, Sung-Jin,Shiwani, Supriya,Singh, Naresh Kumar Asian Australasian Association of Animal Productio 2020 Animal Bioscience Vol.33 No.2
Objective: Orchastric changes in the mammary glands are vital, especially during lactation. The secretary epithelial cells together with the supporting myoepithelial and stromal cells function cordially to secrete milk. Increase in the number of luminal epithelial cells and a decrease in adipocytes are visible during lactation, whereas the reverse happens in the involution. However, an early involution occurs if the epithelial cells transdifferentiate towards adipocytes during the lactation period. We aimed to inhibit the adipocyte transdifferentiation of luminal cells by restraining the peroxisomal proliferator-activated receptor γ (PPARγ) pathway. Methods: Linolenic acid (LA) and thiazolidinediones (TZDs) induced adipogenesis in mammary epithelial cells were conducted in monolayer, mixed culture as well as in transwell plate co-culture with mammary myoepithelial cells. Results: Co-culture with myoepithelial cells showed higher adipogenic gene expression in epithelial cells under LA+TZDs treatment. Increase in the expressions of PPARγ, CCAAT/enhancer-binding protein α and vimentin in both mRNA as well as protein levels were observed. Whereas, bisphenol A diglycidyl ether treatment blocked LA+TZDs induced adipogenesis, as it could not show a significant rise in adipose related markers. Although comparative results were found in both mixed culture and monolayer conditions, co-culture technic was found to work better than the others. Conclusion: Antagonizing PPARγ pathway in the presence of myoepithelial cells can significantly reduce the adipogenisis in epithelial cells, suggesting therapeutic inhibition of PPARγ can be considered to counter early involution or excessive adipogenesis in mammary epithelium in animals.