Expression of Putative Stem Cell Markers Related to Developmental Stage of Sheep Mammary Glands

It is thought that the regenerative capacity of the mammary gland following post-lactation involution resides in multipotent stem cells within the luminal tissue. Adult stem cells make up a small percentage of the cells found in mature organ systems, however to define useful markers has long been a challenge. c-Kit (KIT) and its ligand stem cell factor (KITLG), ATP-binding cassette sub-family G member 2 (ABCG2) and Musashi 1 (MSI1) are good candidate to identify progenitor cells in their niche. Using real-time PCR we showed that KIT, KITLG and MSI1 expressions were up regulated before lambing and at involution relatively to prepubertal stage. The in situ hybridization analysis for KIT gene confirmed and localized the expression in luminal epithelial cells. The changes in the expression profile of putative stem cell markers in mammary glands of sheep suggest that they modify with the progression of lactation cycle, being up regulated during differentiation and down regulated during lactation

Hydroxytyrosol, an ingredient of olive oil, reduces triglyceride accumulation and promotes lipolysis in human primary visceral adipocytes during differentiation

Hydroxytyrosol has various pharmacological properties, including anti-oxidative stress and anti-inflammatory activities, preventing hyperglycemia, insulin resistance, and the metabolic syndrome. The present study is focused on the anti-adipogenic and lipolytic activity of hydroxytyrosol on primary human visceral adipocytes. Pre-adipocytes were analyzed after 10 (P10) and 20 (P20) days of treatment during differentiation and after 7 (A7) days of treatment when they reached mature shape. The treatment with hydroxytyrosol extract significantly (P\u2009<\u20090.001) increased apoptosis in P10 and P20 cells in comparison to control and A7 cells; significantly (P\u2009<\u20090.001) reduced triglyceride accumulation in P20 cells compared to P10 and control cells; and significantly (P\u2009<\u20090.001) increased lipolysis in P20 cells in comparison to control cells and A7 mature adipocytes. Hydroxytyrosol-treated P20 cells significantly (P\u2009<\u20090.05) increased expression of genes involved in inhibition of adipogenesis, such as GATA2, GATA3, WNT3A, SFRP5, HES1, and SIRT1. In contrast, genes involved in promoting adipogenesis such as LEP, FGF1, CCND1, and SREBF1 were significantly down-regulated by hydroxytyrosol treatment. These data suggest that hydroxytyrosol promotes lipolysis and apoptotic activity in primary human visceral pre-adipocytes during differentiation and does not affect already mature adipocytes. \ua9 2016 by the Society for Experimental Biology and Medicine

Different anti-adipogenic effects of bio-compounds on primary visceral pre-adipocytes and adipocytes.

Several natural compounds exhibit strong capacity for decreasing triglyceride accumulation, enhancing lipolysis and inducing apoptosis. The present study reports the anti-adipogenic effects of Silybum marianum (SL), Citrus aurantium (CA), Taraxacum officinale (TO), resveratrol (RE), Curcuma longa (CU), caffeine (CF), oleuropein (OL) and docosahexaenoic acid (DHA) in reducing differentiation and increasing lipolysis and apoptosis. Analyses were performed on human primary visceral pre-adipocytes after 10 (P10) and 20 (P20) days of treatment during differentiation and on mature adipocytes after 7 days of treatment (A7). The percentage of apoptosis induced by TO extract in P10 and P20 cells was significantly higher than that induced by all other compounds and in CTRL cells. Triglyceride accumulation was significantly lower in cells treated with DHA, CF, RE in comparison to cells treated with OL and in CTRL cells. Treatments with CF, DHA and OL significantly incremented lipolysis in P20 cells in comparison to other compounds and in CTRL cells. On the contrary, the treatment of A7 cells with OL, CA and TO compounds significantly increased cell lipolysis. The addition of CF in differentiating P20 pre-adipocytes significantly increased the expression of genes involved in inhibition of adipogenesis, such as GATA2, GATA3, WNT1, WNT3A, SFRP5, and DLK1. Genes involved in promoting adipogenesis such as CCND1, CEBPB and SREBF1 were significantly down-regulated by the treatment. The screening of bioactive compounds for anti-adipogenic effects showed that in differentiating cells TO extract was the most effective in inducing apoptosis and CF and DHA extracts were more efficient in inhibition of differentiation and in induction of cell lipolysis

Effects of Two Different Rhodiola rosea Extracts on Primary Human Visceral Adipocytes

Rhodiola rosea (Rro) has been reported to have various pharmacological properties, including anti-fatigue, anti-stress and anti-inflammatory activity. It is also known to improve glucose and lipid metabolism, but the effects of Rhodiola rosea on adipocyte differentiation and metabolism are not still elucidated. In this study the anti-adipogenic and lipolytic activity of two extracts of Rhodiola rosea, containing 3% salidroside (RS) or 1% salidroside and 3% rosavines (RR) on primary human visceral adipocytes was investigated. Pre-adipocytes were analyzed after 10 and 20 days of treatment during differentiation and after 7 days of treatment when they reached mature shape. The RS extract significantly induced higher apoptosis and lipolysis in comparison to control cells and to RR extract. In contrast, RR extract significantly reduced triglyceride incorporation during maturation. Differentiation of pre-adipocytes in the presence of RS and RR extracts showed a significant decrease in expression of genes involved in adipocyte function such as SLC2A4 and the adipogenic factor FGF2 and significant increase in expression of genes involved in inhibition of adipogenesis, such as GATA3, WNT3A, WNT10B. Furthermore RR extract, in contrast to RS, significantly down-regulates PPARG, the master regulator of adipogenesis and FABP4. These data support the lipolytic and anti-adipogenetic activity of two different commercial extracts of Rhodiola rosea in primary human visceral pre-adipocytes during differentiation

Effect of plant extracts on H2O2-induced inflammatory gene expression in macrophages

Background: Arctium lappa (AL), Camellia sinensis (CS), Echinacea angustifolia, Eleutherococcus senticosus, Panax ginseng (PG), and Vaccinium myrtillus (VM) are plants traditionally used in many herbal formulations for the treatment of various conditions. Although they are well known and already studied for their anti-inflammatory properties, their effects on H2O2-stimulated macrophages are a novel area of study. Materials and methods: Cell viability was tested after treatment with increasing doses of H2O2 and/or plant extracts at different times of incubation to identify the optimal experimental conditions. The messenger (m)RNA expression of TNF\u3b1, COX2, IL1\u3b2, NF\u3baB1, NF\u3baB2, NOS2, NFE2L2, and PPAR\u3b3 was analyzed in macrophages under H2O2 stimulation. The same genes were also quantified after plant extract treatment on cells pre-stimulated with H2O2. Results: A noncytotoxic dose (200 \u3bcM) of H2O2 induced active mRNA expression of COX2, IL1\u3b2, NFE2L2, NF\u3baB1, NF\u3baB2, NOS2, and TNF\u3b1, while PPAR\u3b3 was depressed. The expression of all genes tested was significantly (P,0.001) regulated by plant extracts after pre-stimulation with H2O2. COX2 was downregulated by AL, PG, and VM. All extracts depressed IL1\u3b2 expression, but upregulated NFE2L2. NF\u3baB1, NF\u3baB2, and TNF\u3b1 were downregulated by AL, CS, PG, and VM. NOS2 was inhibited by CS, PG, and VM. PPAR\u3b3 was decreased only after treatment with E. angustifolia and E. senticosus. Conclusion: The results of the present study indicate that the stimulation of H2O2 on RAW267.4 cells induced the transcription of proinflammatory mediators, showing that this could be an applicable system by which to activate macrophages. Plant extracts from AL, CS, PG, and VM possess in vitro anti-inflammatory activity on H2O2-stimulated macrophages by modulating key inflammation mediators. Further in vitro and in vivo investigation into molecular mechanisms modulated by herbal extracts should be undertaken to shed light on the development of novel modulating therapeutic strategies

Dynamic of lipid droplets and gene expression in response to beta-aminoisobutyric acid treatment on 3T3-L1 cells

Research on adipobiology has recognized the browning process of white adipocytes as a potential therapeutic strategy for the treatment of obesity and related morbidities. Physical exercise stimulates the secretion of myokines, such as \uf062-aminoisobutyric acid (BAIBA), which in turn promotes adaptive thermogenesis. White adipocyte conversion to brown cells involves dynamic changes in lipid droplet (LD) dimension and in the transcription of brown-specific marker genes. This study analyzes the effect of different doses of BAIBA and at different days of development on 3T3-L1 cells by evaluating morphological changes in LDs and the expression of browning gene markers. Results suggested that the highest concentration of BAIBA after 4 days of differentiation produced the most significant effects. The number of LDs per cell increased in comparison to control cells, whereas the surface area significantly decreased. Brown adipocyte markers were up-regulated, but the effect of treatment was lost at 10 days of differentiation. The thermogenic program induced by BAIBA may reflect a rapid adaptation of adipose tissue to physical exercise. This connection stresses the beneficial impact of physical exercise on metabolic health

Expression of NGF, BDNF and their receptors in subcutaneous adipose tissue of lactating cows

Currently, there are no reports of neurotrophins in adipose tissue of cows. The distribution of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and their high-affinity tyrosine kinase receptors TrkA and TrkB, was investigated by immunohistochemical method in the subcutaneous adipose tissue of cow at mid-lactation. Results revealed the localization of NGF and BDNF along the plasma membrane and cytoplasm of adipocytes. Neurotrophin receptors TrkA and TrkB showedmoderate and strong positive staining in adipocytes, respectively. The expression of NGF, BDNF, TRKB \u2014 but not of TRKA \u2014 was also confirmed at transcriptional level by RT-PCR analyses. Considering the involvement of BDNF on fat metabolism and of NGF on activation of the sympathetic response in human and rodents, these neurotrophins could be related to lipogenesis and lipolysis occurring during lactation in cows. The local production of these neurotrophins supports their potential paracrine function for the regulation of adipocyte activity and deserve further investigations

Expression profile of caseins, estrogen and prolactin receptors in mammary glands of dairy ewes

In this study, we analyzed the developmental expression of estrogen receptors (ESR1 and ESR2), prolactin receptors (PRLR) and casein genes (CSN1S1, CSN1S2, CSN2 and CSN3) in the ewe mammary glands from prepubertal stage to involution. Using Real-time PCR we showed that the activation of casein genes transcription was up regulated during lactation and significantly down regulated before lambing and at involution in comparison to the expression measured in the prepubertal group. The highest expression of ESR1 and ESR2 genes occurred in prepubertal group compared to adult group. The PRLR expression of the short and long forms was up regulated before lambing and down regulated during lactation and involution. Thus, the mRNA expression data for ESRs and PRLR show clear regulatory changes suggesting involvement of these receptors in sheep mammary glands during development to involution. Casein genes transcription could be primed through PRLR signal, but other factors may be necessary for milk protein longterm expression during lactation

Nutrigenomic activity of plant derived compounds in health and disease: Results of a dietary intervention study in dog

The study was conducted to investigate the effects of dietary administrations of four nutraceuticals in dogs. Seventy four dogswere enrolled in the trials, 24 healthy dogs were fedwith a control diet (CT) and the experimental groups received for 60 days the same diet supplemented with nutraceuticals, namely Echinacea angustifolia (EA, 0.10 mg/kg live weight as echinacoside; 14 dogs), Vaccinium myrtillus (VM, 0.20 mg/kg live weight as anthocyanidin, 13 dogs), Curcuma longa (CL, 6.60 mg/kg live weight as curcumin, 18 dogs with arthrosis), and Sylibum marianum (SM, 1.5 mg/kg live weight as sylibin, 8 dogs with hepatopathy). Dogs were weighted at the beginning of study and blood samples were collected at the beginning (T0) and at the end (T60) of the study. VM significantly down regulated TNF, CXCL8, NFKB1 and PTGS2 and decreased plasma ceruloplasmin (CuCp). The activity of EA was evidenced by the significant decrease of TNF and NFKB1 expression and CuCp levels and by the increase of plasma Zn. Administration of CL caused a significant decrease of CuCp and increase of Zn and a down regulation of TNF, CXCL8, NFKB1 and PTGS2, corroborating the anti-inflammatory action of curcuminoids. After 60 days of treatmentwith SM, plasma ALT/GPT activitywas reduced and paraoxonase was increased, supporting the antioxidant activity of silymarin, also confirmed by the significant up regulation of SOD2. Results indicated that nutraceutical administrations in dogs can be an interesting approach to modulate immune response in order to improve health condition of animals