Effects of L-Leu-L-Leu peptide on growth, proliferation, and apoptosis in broiler intestinal epithelial cells.

Small peptides are nutrients and bioactive molecules that have dual regulatory effects on nutrition and physiology. They are of great significance for maintaining the intestinal health and production performance of broilers. We here cultured the primary small intestinal epithelial cells (IEC) of chicken in a medium containing L-Leu (Leu) and L-Leu-L-Leu (Leu-Leu) for 24 h. The untreated cells were considered as the control group. The growth, proliferation, and apoptosis of IEC were examined. By combining RNA-seq and label-free sequencing technology, candidate genes, proteins, and pathways related to the growth, proliferation, and apoptosis of IEC were screened. Immunofluorescence detection revealed that the purity of the isolated primary IEC was >90%. The Leu-Leu group significantly promoted IEC growth and proliferation and significantly inhibited IEC apoptosis, and the effect was better than those of the Leu and control groups. Using transcriptome sequencing, four candidate genes, CCL20, IL8L1, IL8, and IL6, were screened in the Leu group, and one candidate gene, IL8, was screened in the Leu-Leu group. Two candidate genes, IL6 and RGN, were screened in the Leu-Leu group compared with the Leu group. Nonquantitative proteomic marker sequencing results revealed that through the screening of candidate proteins and pathways, found one growth-related candidate protein PGM3 and three proliferation-related candidate proteins RPS17, RPS11, and RPL23, and two apoptosis-related candidate proteins GPX4 and PDPK1 were found in the Leu-Leu group compared with Leu group. In short, Leu-Leu could promote IEC growth and proliferation and inhibit IEC apoptosis. On combining transcriptome and proteome sequencing technologies, multiple immune- and energy-related regulatory signal pathways were found to be related to IEC growth, proliferation, and apoptosis. Three candidate genes of IL8, IL6, and RGN were identified, and six candidate proteins of PGM3, RPS17, RPS11, RPL23, GPX4, and PDPK1 were involved in IEC growth, proliferation, and apoptosis. The results provide valuable data for preliminarily elucidating small peptide-mediated IEC regulation pathways, improving the small peptide nutrition theoretical system, and establishing small peptide nutrition regulation technology.

Analysis of the antioxidant activity of toons sinensis extract and their biological effects on broilers.

Plant extracts are rich in a variety of nutrients and contain a large number of bioactive compounds, and compared with traditional feed additives, they have advantages such as wide sources, natural safety and rich nutrition. This study employed in vitro antioxidant and animal experiments to comprehensively evaluate the use of Toona sinensis extract (TSE) in broiler production. 508 1-day-old Cobb 500 broilers were randomly assigned to the 7 experimental groups with 6 replications and 12 birds/replicate. Two groups received Vitamin C (VC) 300 g/t and Vitamin E 500 g/t, and five dose groups of TSE received 0, 300, 600, 900, and 1,200 g/t of TSE in their feed. The study spanned 42 days, with a starter phase (1-21 days) and a finisher phase (22-42 days). The results showed that compared to ascorbic acid, TSE had the scavenging ability of 2,2-Diphenyl-1-picrylhydrazyl and hydroxyl radical, with IC50 values of 0.6658 mg/mL and 33.1298 mg/mL, respectively. Compared to TSE 0 group, broilers fed with 1,200 g/t TSE showed significant weight gain during the starter phase and increased the feed-to-weight gain ratio during both the starter and finisher phases. Additionally, broilers receiving 1,200 g/t TSE had enhanced dry matter and organic matter utilization. Concerning meat quality, broilers in the 1,200 g/t TSE group demonstrated increased cooked meat yield, and pH value, as well as higher antioxidant capacity (T-AOC), dismutase (SOD), and glutathione peroxidase (GSH-PX) in serum. In addition, there was no significant difference in ileal microflora due to TSE supplementation. In summary, this study confirms the positive impact of a dietary inclusion of 1,200 g/t TSE on broiler growth, meat quality, and serum antioxidants.

[Down-regulation of pannexin 2 channel enhances cisplatin-induced apoptosis in testicular cancer I-10 cells].

OBJECTIVE: To investigate the effect of down-regulation of pannexin 2 (Panx-2) channels on cisplatin-induced apoptosis in I-10 cells. METHODS: The expression of Panx-2 protein in testicular cancer cells was detected with Western blotting. The testicular cancer cell line I-10 was transfected with two short hairpin RNA (shRNA1 and shRNA2) via Lipofectamine2000, the empty vector (NC group) or Lipofectamine2000 (blank control group), and the changes in the expression of Panx-2 was detected with Western blotting. The effects of transfection with a Panx-2 inhibitor on surviving fraction of the cells treated with cisplatin (16 µmol/L) for 24 h, 48 h and 72 h was assessed with MTT assay, and the clonogenic capacity of the cells was evaluated with colony-forming assay. At 8 h after incubation with 16 µmol/L cisplatin, AnnexinV/PI double staining was used to detect the early apoptosis of the cells. After 24 h of treatment with 16 µmol/L cisplatin, the cells were examined for expressions of caspase-3, Bcl-2 and Bax using Western blotting. RESULTS: The expression of Panx-2 was significantly increased in cisplatin-resistant I-10/DDP (P < 0.001) cells and Tcam-2/DDP (P < 0.01) cells as compared with I-10 cells and Tcam-2 cells. Transfection of I-10 cells with shRNA1 and shRNA2 resulted in significantly decreased Panx-2 expression (P < 0.05) and significantly reduced cell surviving fraction (P < 0.001). In the presence of cisplatin, the cells in NC group showed a higher clonogenic efficiency than those in shRNA1 and shRNA2 groups (P < 0.001). The early-stage apoptosis rate of the cells in shRNA1 and shRNA2 groups were significantly higher than that in NC group (P < 0.01). Panx-2 knockdown in I-10 cells significantly increased caspase-3 and Bax expressions (P < 0.05) and significantly decreased the expression of Bcl-2 (P < 0.01). CONCLUSIONS: Down-regulation of Panx-2 channel enhances cisplatin-induced apoptosis in cultured testicular cancer cells.