This study examined the influence of chlorogenic acid (CGA) on gut antioxidant status, morphology, barrier function, immunity, and cecal microbiota in late-peak laying hens. A total of 240 Hy-Line Brown hens, aged 43 weeks, were randomly assigned to four groups, the basal diet +0, 400, 600, and 800 mg/kg CGA, for 12 weeks. The results revealed that CGA significantly reduced ileal H₂O₂ and malondialdehyde levels; increased duodenal height, ileal villus height, and villus height-to-crypt depth ratio; while decreasing jejunal crypt depth. The 600 and 800 mg/kg CGA significantly upregulated the duodenal, jejunal, and ileal ZO-1 and occludin gene expression; increased IgG levels in serum and ileum; and upregulated ileal IgA gene expression. The 600 mg/kg CGA significantly upregulated CD3D and CD4 gene expression, while downregulating IL-1β gene expression in duodenum, jejunum, and ileum. Moreover, CGA changed the gut microbiota structure. The SCFA-producing bacteria unclassified_f__Peptostreptococcaceae, unclassified_f_Oscillospiraceae, Pseudoflavonifractor, Lachnospiraceae_FCS020_group, Oscillospira, Elusimicrobium, Eubacterium_ventriosum_group, Intestinimonas, and norank_f_Coriobacteriales_Incertae_Sedis were significantly enriched in the 400, 600, and/or 800 mg/kg CGA groups. The bacteria Lactobacillus, Bacillus, and Akkermansia were significantly enriched in the 600 mg/kg CGA group. Conclusively, dietary CGA (600–800 mg/kg) improved intestinal antioxidant status, morphology, barrier and immune function, and beneficial microbiota growth in late-peak laying hens.

This research examined the impact of chlorogenic acid (CGA) on laying performance, antioxidant capacity, egg quality, hepatic inflammation, mitochondrial function, and lipid metabolism in hens subjected to hydrogen peroxide (H₂O₂)-induced oxidative stress (OS). Three hundred sixty healthy 43-wk-old Hy-Line brown hens were randomly assigned to six treatments: a basal diet + 0 (control and H₂O₂), 600 (600 mg/kg CGA and 600 mg/kg CGA + H₂O₂), and 800 (800 mg/kg CGA and 800 mg/kg CGA + H₂O₂) mg/kg CGA for 84 d. On the 64th and 78th days of the trial, hens in groups H₂O₂, 600 mg/kg CGA + H₂O₂, and 800 mg/kg CGA + H₂O₂ were injected intraperitoneally with 10% H₂O₂. The results demonstrated that 600 and 800 mg/kg CGA significantly improved the egg production rate (EPR) and egg quality and reduced lipid peroxidation compared to the control group. The 800 mg/kg CGA showed greater improvements in the EPR and average egg weight (AEW) compared to the 600 mg/kg dose. Conversely, H₂O₂ exposure significantly decreased the EPR, AEW, and egg quality and increased feed conversion rate and average daily feed intake. H₂O₂ exposure significantly decreased serum T-AOC and increased serum MDA levels while reducing hepatic T-SOD, GSH-Px, and CAT activities. Meanwhile, H₂O₂ exposure significantly elevated liver reactive oxygen species levels, pathological damage, and NF-κB, TNFα, and IL-1β gene expression. Additionally, H₂O₂ treatment disrupted hepatocyte mitochondrial structure and significantly increased the expression of VDAC1 protein, and IP3R, GRP75, MCU, Fis1, and MFF genes, while downregulating the expression of MFN2 protein and PGC1α gene. Oil Red O staining demonstrated that H₂O₂ induced significant lipid accumulation in hepatocytes. Concurrently, H₂O₂ significantly increased serum triglycerides, total cholesterol, and liver triglycerides levels while decreasing serum hepatic lipase activity. This was primarily attributed to the significant upregulation of liver SREBP1, FASN, and ACC genes and the downregulation of the liver CPT1 gene induced by H₂O₂. Furthermore, CGA pretreatment effectively prevented the degeneration in laying performance and egg quality, as well as OS, liver inflammation, pathological damage, and mitochondrial dysfunction induced by H₂O₂. CGA inhibited H₂O₂-induced hepatic lipid accumulation by upregulating fatty acid oxidation-related gene expression and downregulating fatty acid synthesis-related gene expression. These findings indicate that the dietary addition of 800 mg/kg of CGA is the optimum supplementation dose. CGA can enhance laying performance and egg quality while alleviating OS, hepatic inflammation, mitochondrial dysfunction, and lipid accumulation in H₂O₂-challenged laying hens.

Cadmium (Cd) is known as a female reproductive toxicant. Our previous study has shown that Cd can influence the proliferation and cell cycle of granulosa cells and induce apoptosis. MicroRNAs (miRNAs) play an important role in the regulation of Cd-induced granulosa cell damage in chickens. However, the mechanism remains unclear. In this study, we investigated the mechanisms by which microRNA-129-1-3p (miR-129-1-3p) regulates Cd-induced cytotoxicity in chicken granulosa cells. As anticipated, exposure to Cd resulted in the induction of oxidative stress in granulosa cells, accompanied by the downregulation of antioxidant molecules and/or enzymes of Nrf2, Mn-SOD, Cu-Zn SOD and CAT, and the upregulation of Keap1, GST, GSH-Px, GCLM, MDA, hydrogen peroxide and mitochondrial reactive oxygen species (mtROS). Further studies found that Cd exposure causes mitochondrial calcium ions (Ca²⁺) overload, provoking mitochondrial damage and apoptosis by upregulating IP3R, GRP75, VDAC1, MCU, CALM1, MFF, caspase 3, and caspase 9 gene and/or protein expressions and mitochondrial Ca²⁺ levels, while downregulating NCX1, NCLX and MFN2 gene and/or protein expressions and mitochondrial membrane potential (MMP). The Ca²⁺ chelator BAPTA-AM or the MCU inhibitor MCU-i4 significantly rescued Cd-induced mitochondrial dysfunction, thereby attenuating apoptosis. Additionally, a luciferase reported assay and western blot analysis confirmed that miR-129-1-3p directly target MCU. MiR-129-1-3p overexpression almost completely inhibited protein expression of MCU, increased the gene and protein expressions of NCLX and MFN2 downregulated by Cd, and attenuated mitochondrial Ca²⁺ overload, MMP depression and mitochondria damage induced by Cd. Moreover, the overexpression of miR-129-1-3p led to a reduction in mtROS and cell apoptosis levels, and a suppression of the gene and protein expressions of caspase 3 and caspase 9. As above, these results provided the evidence that IP3R-MCU signaling pathway activated by Cd plays a significant role in inducing mitochondrial Ca²⁺ overload, mitochondrial damage, and apoptosis. MiR-129-1-3p exerts a protective effect against Cd-induced granulosa cell apoptosis through the direct inhibition of MCU expression in the ovary of laying hens.

Oxidative stress is a well-known challenge in poultry production. While chlorogenic acid (CGA) is recognized for its antioxidant, anti-inflammatory, antibacterial, and lipid-lowering properties, its protective role against oxidative stress-induced intestinal damage in laying hens has not been extensively explored. This study investigated the effects of dietary CGA on the intestinal structure, barrier integrity, and cecal microbiota in laying hens exposed to hydrogen peroxide (H₂O₂). A sum of 240 Hy-Line Brown hens (43-wk-old) was divided into 4 groups (n = 6 replicates/group, 10 hens/replicate). Groups were fed a basal diet with CGA at 0 mg/kg (control, H₂O₂) or 600 mg/kg (600 mg/kg CGA, 600 mg/kg CGA + H₂O₂) for 12 weeks. Intraperitoneal injections of 10 % H₂O₂ were performed on days 64 and 78 in the H₂O₂ and 600 mg/kg CGA + H₂O₂ groups. The results showed that CGA pretreatment prevented the H₂O₂-induced oxidative stress, as evidenced by reducing MDA and H₂O₂ levels, along with enhanced activities of T-SOD, CAT, and GSH-Px (P < 0.05). CGA administration significantly countered the decline in productivity, villus height (VH), and the VH-to-crypt depth (CD) ratio, as well as the increase in CD caused by H₂O₂ (P < 0.05). Furthermore, CGA ameliorated the adverse effects of H₂O₂ on intestinal barrier function, inflammation, and immune response, including preventing H₂O₂-induced downregulation of occludin, TGFβ, IL-10, IgA, cluster of differentiation 3D (CD3D), and CD4 gene expression, and upregulation of TLR4 and TNFα genes and MyD88 protein expression in the gut tissues. Additionally, H₂O₂ was found to disrupt the structural composition of the cecal microbiota, resulting in an increased relative abundance of potential pathogenic taxa such as unclassified_f_Synergistaceae and uncultured_bacterium_Treponema. This disruption was accompanied by a decrease in the relative abundance of potentially beneficial bacterial species, including Bacteroides_caecicola, Collinsella_massiliensis_g_collinsella, unclassified_g_Oscillibacter, and uncultured_bacterium_g_Turicibacter. Furthermore, elevated serum levels of diamine oxidase (DAO) and lipopolysaccharide (LPS) were observed. More importantly, pre-treatment with CGA ameliorated the microbiota structure and serum DAO and LPS levels in laying hens subjected to H₂O₂. Our findings collectively indicate that CGA may mitigate H₂O₂-induced oxidative stress, barrier dysfunction, and immune impairment, while enhancing egg production and gut microbiota in laying hens. These findings highlight the novel role of CGA in mitigating oxidative stress-induced intestinal damage and microbiota dysbiosis in laying hens, providing new insights into its potential as a feed additive for improving poultry health and productivity.

Background: in the twenty-first century, the emergence of COVID-19 as a highly transmissible pandemic disease caused by SARS-CoV-2 posed a significant threat to humanity. Aims & Objectives: the disease spreads through small respiratory droplets, necessitating the use of various compounds for treatment, with alkaloids being recognized as particularly crucial owing to their diverse pharmaceutical properties. Methodology: in this study, a dataset comprising 100 natural alkaloids obtained from the literature was transformed into 2D chemical structures using Chem Draw 19.1. Subsequently, 3DQSAR studies were conducted on the dataset, resulting in the automatic screening of 50 compounds from the initial pool of 100 compounds. The values of q² and r² of the validated field-based 3DQSAR model were 0.7186 and 0.971, respectively. The validated atom-based 3DQSAR model has q² and r² scores of 0.6025 and 0.9845, respectively. Based on the obtained results, 10 compounds with exceptionally active predictive IC₅₀ values were selected for further analysis. Docking experiments were then performed on the selected compounds, and the top three compounds with the highest docking scores were identified as diazepinomicin, (+)-N-methylisococlaurine, and hymenocardine-H. After docking, MM–GBSA was performed on the complexes of diazepinomicin, (+)-N-methylisococlaurine and hymenocardine-H with their corresponding proteins, which resulted in the authentication of the molecular docking scores. MD simulations were also performed to check the flexibility, stability and compactness of these complexes for revalidation of docking scores. Results: finally, ADMET experiments revealed that (+)-N-methylisococlaurine exhibited the most favourable properties among these three compounds.