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Inhibitory Regulation of Chicken Mx against Avian Viruses in Chicken Spermatogonial Stem Cells
Ju-Young Ji,Kuppannan Gobianand,Jong-Ju Park,Jin-Gu No,Ju Sung Yang,Man Sung Park,Dong Kee Jeong,Dong-Hoon Kim,Jin Ki Park,Sung-June Byun,Chang Sun Song,Jae Gyu Yoo 한국동물생명공학회(구 한국동물번식학회) 2013 Reproductive & Developmental Biology(Supplement) Vol.37 No.2s
Mx is regulated by type I interferons and contains a typical GTP-binding motif like other members of the GTPase dynamin family. However, the functions and working mechanisms of the Mx protein in chicken spermatogonial stem cells (SSCs) are not well documented. In the present study, Mx-overexpressing chicken SSCs (chMx-SSCs) were established and the antiviral activity of chMx-SSCs against Newcastle disease virus, avian influenza viruses was investigated in vitro. For chicken SSCs isolation, day 20 fetal males derived testes were initially subjected to digestion by collagenase IV followed by 0.25% trypsin–EDTA. After discarding the supernatant, the cells were cultured in SSC medium. SSC colonies expressed pluripotent markers such as stagespecific embryonic antigen-1, Oct-4, Nanog, and Sox-2. Chicken Mx gene was constructed in plasmid DNA vector (pcDNA3.1/V5-His A-chMX) and ChMx-SSCs lines were established with chMX constructs. The antiviral activity of ChMx-SSCs was determined by real-time RT-PCR, flow cytometry, and western blot analyses after infection with Newcastle disease virus-green fluorescent protein (GFP) and avian influenza viruses (H9N2 and H1N1). ChMx-SSCs inhibited recombinant Newcastle disease virus (rNDV)- GFP replication as determined by the calculation of the proportion GFP signal- positive cells by FACS analysis. When SSCs showed 100% GFP expression, chMx- SSCs had only 3.6% GFP expression. At 24 h after avian influenza virus infection, chMx-SSCs had a lower hemagglutinin protein level and a higher level of Mx protein. When the number of released virion particles was estimated by plaque-formation assay, chMx-SSCs had significantly fewer stained visible plaques in the MDCK layer than SSCs. Our results suggest that overexpression of chicken Mx directly stimulates antiviral activity resulting in downregulation of viral progeny release. Chicken Mx overexpression in chicken SSCs can be applied for the production of virus resistant transgenic chicken via direct transplantation of chMx-SSCs into the testis.
Ibrahim, Muhammed,Jang, Mi,Park, Mina,Gobianand, Kuppannan,You, Seungkwon,Yeon, Sung-Heom,Park, Sungkwon,Kim, Min Ji,Lee, Hyun-Jeong The Royal Society of Chemistry 2015 Food & function Vol.6 No.7
<P>Obesity is a global health problem that requires the utmost attention. Apart from other factors the trans-differentiation of mesenchymal stem cells (MSCs) into adipocytes is an added detrimental factor causing the intensification of obesity. The main objective of this present study is to analyse whether capsaicin is capable of inhibiting the differentiation of BMSCs to adipocytes. Bone marrow mesenchymal stem cells (BMSCs) were obtained and exposed to different concentrations of capsaicin for a period of 6 days following 2 days of adipogenic induction. The capsaicin exposed cells were collected at three different time points (2, 4 and 6 days) and subjected to various analyses. BMSCs after exposure to capsaicin showed dose and time dependent reduction in cell viability and proliferation. Interestingly, capsaicin induced cell cycle arrest at G0-G1and increased apoptosis by increasing reactive oxygen species (ROS) and reactive nitrogen species (RNS) production. Capsaicin significantly inhibited the early adipogenic differentiation, lipogenesis and maturation of adipocytes with concomitant repression of PPARγ, C/EBPα, FABP4 and SCD-1. Taken together, the results of the present study have clearly emphasized that capsaicin potentially inhibits the adipogenic differentiation of mesenchymal stem cells<I>via</I>many different pathways (anti-proliferative, apoptotic and cell cycle arrest) through the stimulation of ROS and RNS production. Thus, capsaicin not only suppresses the maturation of pre-adipocytes into adipocytes but also inhibits the differentiation of mesenchymal stem cells into adipocytes.</P>
Effects of Capsaicin on Adipogenic Differentiation in Bovine Bone Marrow Mesenchymal Stem Cell
Jeong, Jin Young,Suresh, Sekar,Park, Mi Na,Jang, Mi,Park, Sungkwon,Gobianand, Kuppannan,You, Seungkwon,Yeon, Sung-Heom,Lee, Hyun-Jeong Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.12
Capsaicin is a major constituent of hot chili peppers that influences lipid metabolism in animals. In this study, we explored the effects of capsaicin on adipogenic differentiation of bovine bone marrow mesenchymal stem cells (BMSCs) in a dose- and time-dependent manner. The BMSCs were treated with various concentrations of capsaicin (0, 0.1, 1, 5, and $10{\mu}M$) for 2, 4, and 6 days. Capsaicin suppressed fat deposition significantly during adipogenic differentiation. Peroxisome proliferator-activated receptor gamma, cytosine-cytosine-adenosine-adenosine-thymidine/enhancer binding protein alpha, fatty acid binding protein 4, and stearoyl-CoA desaturase expression decreased after capsaicin treatment. We showed that the number of apoptotic cells increased in dose- and time-dependent manners. Furthermore, we found that capsaicin increased the expression levels of apoptotic genes, such as B-cell lymphoma 2-associated X protein and caspase 3. Overall, capsaicin inhibits fat deposition by triggering apoptosis.