Effect of Flammulina velutipes polysaccharides on endoplasmic reticulum stress-mediated apoptosis by activating PLC-IP3 pathway in HepG2 cells.

Flammulina velutipes polysaccharides (FVP) have been proven to induce apoptosis in HepG2 cells. It is well known that endoplasmic reticulum stress (ERS) is involved in apoptosis. However, ERS mediates FVP-induced apoptosis in HepG2 cells remains unclear. In our study, the results indicated that FVP caused ERS in HepG2 cells. They showed that FVP were water-soluble polysaccharides with the weight average molecular weight of 1972 kDa, which were mainly composed of mannose, gluconic acid, glucose, galactose, xylose and fructose in a molar ratio of 6.6 : 1.3 : 79.9 : 7.4 : 3.4 : 1.5. After FVP treatment, the expression levels of genes and proteins related to ERS were upregulated. The inhibition of ERS by 4-phenylbutyric acid (4-PBA) pretreatment could significantly reduce the role of FVP in inducing apoptosis. We further found the results of immunofluorescence and flow cytometry showing that Ca2+ in the ERS leaked out, and the intracellular Ca2+ concentration increased after FVP treatment. The pretreatment with the phospholipase C (PLC) inhibitor U73122 proved that FVP caused excessive intracellular Ca2+ concentration by activating the phospholipase C-inositol-1,4,5-triphosphate (PLC-IP3) pathway, resulting in ERS, and ultimately leading to apoptosis. In summary, our results indicated that FVP induced ERS-mediated apoptosis by activating PLC-IP3 pathway in HepG2 cells. PRACTICAL APPLICATION: This work may suggest that FVP could be used as an adjuvant therapy to anticancer drugs, providing new application prospects and possibilities.

Flammulina velutipes stem regulates oxidative damage and synthesis of yolk precursors in aging laying hens by regulating the liver-blood-ovary axis.

Egg production levels in late laying hens are negatively correlated with increasing age. Decreased liver and ovarian function in aging laying hens is accompanied by decreased antioxidant capacity, reproductive hormone levels, and follicular development, resulting in decreased synthesis of yolk precursors. The golden needle mushroom (Flammulina velutipes) has been reported to exhibit anti-inflammatory, antioxidant, and hypolipidemic properties. We aimed to reveal the therapeutic effects of F. velutipes stem (FVS) on liver-blood-ovary axis and investigate the underlying mechanisms. A total of 360 sixty-seven-wk-old laying hens were randomized into 4 treatment groups: 1) basal maize-soybean meal diet (CON); 2) basal maize + 20 g/kg FVS (2% FVS); 3) basal maize + 40 g/kg FVS (4% FVS); and 4) basal maize + 60 g/kg FVS (6% FVS). FVS groups demonstrated significantly increased egg production and ovarian development compared with the CON group. The addition of FVS increased the levels of antioxidant enzymes (GSH-Px, T-SOD, and T-AOC) in the liver, serum, and ovaries and decreased malondialdehyde levels by regulating the expression of proteins related to the Keap1-Nrf2/ARE signaling pathway. Additionally, FVS significantly decreased ovarian apoptosis by regulating Bax, Bcl-2, and caspase3 mRNA and protein expression levels. FVS significantly increased the expression levels of estradiol, progesterone, luteinizing hormone, and follicle stimulating hormone and their respective receptors. With increased levels of estradiol transported to the liver through the bloodstream, targeted binding to estrogen receptor (ER)-α and ER-ß led to significant increases in ApoVLDL II, ApoB, and VTG II mRNA expression associated with yolk precursor synthesis. FVS decreased the levels of triglyceride and total cholesterol and significantly increased the expression of lipid metabolism, and transport-related mRNAs (FAS, PPAR-a/γ, and MTTP) in the liver. Therefore, the dietary supplementation of FVS can maintain the productive performance of aging laying hens by alleviating the degree of oxidative stress and regulating the transport of functional substances along the liver-blood-ovary axis, thereby improving the synthesis of yolk precursors.

Feed Supplemented with Aronia melanocarpa (AM) Relieves the Oxidative Stress Caused by Ovulation in Peak Laying Hens and Increases the Content of Yolk Precursors.

The continuous ovulation of laying hens during the peak period is likely to cause oxidative stress, resulting in a reduction in the laying cycle over time. The aim of this study was to evaluate the antioxidant effects of Aronia melanocarpa (AM) in the diet and its effect on the yolk precursor content caused by ovulation in laying hens during the peak period. A total of 300 25-week-old Roman brown laying hens were randomly divided into five groups with six replicates in each group, 10 in each replicate. The control group was fed a basal diet, the positive control group was fed a Vitamin C (VC) plus basal diet, and the experimental group was fed 1%, 4%, and 7% doses of AM plus diet according to the principle of energy and nitrogen requirements, which lasted eight weeks. At the end of the study, the egg quality, biochemical, and antioxidant markers, as well as mRNA and protein expressions, were evaluated to determine the potential signaling pathways involved. Results showed that the addition of AM to the feed increased the weight of laying hens at the peak of egg production and improved egg quality. The biochemical markers, as well as the antioxidant parameters in the serum, liver, and ovarian tissues, were ameliorated. The gene and protein expression of recombinant kelch-like ECH-associated protein 1 (Keap1) in the liver and ovarian tissues was decreased, while nuclear factor erythroid-2 related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression was increased. The feed supplemented with AM also increased the estrogen contents and lipid parameters, as well as the gene and protein expressions related to the yolk precursor. Feed supplemented with AM could improve the egg quality and the oxidative stress caused by the ovulation process of laying hens during the peak egg production period by activating the Keap1/Nrf2 signaling pathway. These results suggest that the feed supplemented with 1% AM and 4% AM can improve egg production in peak laying hens.

Aronia melanocarpa (Michx.) Elliott. attenuates dextran sulfate sodium-induced Inflammatory Bowel Disease via regulation of inflammation-related signaling pathways and modulation of the gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Aronia melanocarpa (Michx.) Elliott. Is one of the most functional berries usually used in the preparation of juice and jams, but it has revealed its ethnopharmacological properties due to their richness in biologically active molecules with pharmaceutical and physiological effects. AIMS OF THE STUDY: The aim of this study was to assess the antioxidant and anti-inflammatory effects of Aronia melanocarpa ethanol-extract as well as the possible mechanisms of action involved and the modulation of gut microbiota in Dextran Sulfate Sodium (DSS)-induced Inflammatory bowel disease in mice. MATERIALS AND METHODS: Inflammatory bowel disease (IBD) were induced by DSS in drinking water for 7 days to evaluate the properties of A. melanocarpa ethanol-extract (AME) on the intestinal microflora. AME was administered orally to DSS-induced IBD mice for 21 days. Clinical, inflammatory, histopathological parameters, and different mRNA and proteins involved in its possible mechanism of action were determined as well as gut microbiota analysis via 16S high throughput sequencing. RESULTS: AME improved clinical symptoms and regulated histopathological parameters, pro- and anti-inflammatory cytokines and oxidative stress factors as well as mRNA and protein expressions of transcription factors involved in maintaining the intestinal barrier integrity. In addition, AME also reversed the DSS-induced intestinal dysbiosis effects promoting the production of cecal short chain fatty acids linked to signaling pathways inhibiting IBD. CONCLUSION: AME improved intestinal lesions induced by DSS suggesting that A. melanocarpa berries could have significant therapeutic potential against IBD due to their antioxidant and anti-inflammatory capacities as well as their ability to restore the gut microbiota balance.