Molecular and metabolic responses to immune stress in the jejunum of broiler chickens: transcriptomic and metabolomic analysis.

In the large poultry industry, where farmed chickens are fed at high density, the prevalence of pathogens and repeated vaccinations induce immune stress, which can significantly decrease the production performance and increase the mortality. This study was designed to shed light on the molecular mechanisms and metabolic pathways involved in immune stress through an in-depth analysis of transcriptomic and metabolomic changes in jejunum samples from the broilers. Two groups were established for the experiment: a control group and an LPS group. LPS group received an intraperitoneal injection of LPS solution at a dose of 250 µg per kg at 12, 14, 33, and 35 d of age, whereas the control group received a sterile saline injection. The severity of immune stress was assessed using the Disease Activity Index. A jejunal section was collected to measure the intestinal villus structure (villus length and crypt depth). RNA sequencing and metabolomics data analysis were conducted to reveal differentially expressed genes and metabolites. The results showed that the DAI index was increased and jejunal villus height/crypt depth was decreased in the LPS group. A total of 96 differentially expressed genes and 672 differentially accumulating metabolites were detected in the jejunum by LPS group compared to the control group. The comprehensive analysis of metabolomic and transcriptomic data showed that 23 pathways were enriched in the jejunum and that appetite, nutrient absorption, energy and substance metabolism disorders and ferroptosis play an important role in immune stress in broilers. Our findings provide a deeper understanding of the molecular and metabolic responses in broilers to LPS-induced immune stress, suggesting potential targets for therapeutic strategies to improve the production performance of broiler chickens.

Ginsenoside Rg1 and Re alleviates inflammatory responses and oxidative stress of broiler chicks challenged by lipopolysaccharide.

Previous study showed that ginsenoside Rg1 (Rg1) and ginsenoside Re (Re) alleviated growth inhibition of broiler chicks with immune stress. The aim of this study was to investigate the effect of Rg1 and Re on inflammatory responses, oxidative stress, and apoptosis in liver of broilers with immune stress induced by lipopolysaccharide (LPS). Forty broiler chicks were randomly divided into 4 groups, each group consisting of 10 chickens. The model group, Rg1 group, and Re group were received continuously interval injection of 250 µg/kg body weight LPS at the age of 12, 14, 33, and 35 days to induce immune stress. Control group was injected with an equivalent amount of sterile saline. Then broilers in Rg1 group and Re group were given 1mg/kg body weight Rg1 and Re intraperitoneally 2 h after the LPS challenge respectively. Blood samples were collected for the detection of hormone levels, inflammatory mediators, and antioxidant parameters. Hepatic tissues were taken for pathological observation. Total RNA was extracted from the liver for real-time quantitative polymerase chain reaction analysis. Our results showed that Rg1 or Re could alleviate histological changes of liver, reduce production of stress-related hormones, inhibit inflammatory responses, and enhance antioxidant capacity in broilers challenged by immune stress. In addition, Rg1 or Re treatment upregulated mRNA expression of antioxidant-related genes and downregulated mRNA expression of inflammation-related factors and apoptosis-related genes in the liver of immune-stressed broilers. The results suggest that the plant extracts containing Rg1 and Re can be used for ameliorating hepatic oxidative stress and inflammation and controlling immune stress in broiler chicks.

Hepatic transcriptomics and metabolomics indicated pathways associated with immune stress of broilers induced by lipopolysaccharide.

Broilers with immune stress show decline of growth performance, causing severe economic losses. However, the molecular mechanisms underlying the immune stress still need to be elucidated. One hundred and twenty broiler chicks were randomly assigned to 2 groups with 6 repeats per group, 10 birds per repeat. The model broilers were intraperitoneally injection of 250 µg/kg LPS at 12, 14, 33, and 35 d of age to induce immunological stress. Control group was injected with an equivalent amount of sterile saline. Blood samples from chickens were collected using wing vein puncture at 35 d of age and the serum was obtained for detection of CORT and ACTH. At the end of the experiment, the liver tissues were excised and collected for omics analysis. The results showed that LPS challenge significantly inhibited growth performance, increased relative weight of liver, spleen and decreased relative weight of bursa, as well as enhanced the concentration of serum ACTH and CORT, when compared with the Control. A total of 129 DEGs and a total of 109 differential metabolites were identified between Model and Control group. Transcriptomics profiles revealed that immune stress enhanced the expression of genes related to defense function while declined the expression of genes related to oxidation-reduction process. Metabolomics further suggested that immune stress changed metabolites related to amino acid metabolism, glycerophospholipid metabolism. In addition, integrated analysis suggested that the imbalance of valine, leucine and isoleucine metabolism, glycerophospholipid metabolism, and mTOR signaling pathway played an important role in immune stress of broiler chicks.

Serum metabolomics reveal pathways associated with protective effect of ginsenoside Rg3 on immune stress.

Our previous study has demonstrated that administration of ginsenoside Rg3 ameliorates immune stress by inhibiting inflammatory responses, reducing oxidative damage and upregulating mRNA expression of mTOR, SOD-1, and HO-1. However, the specific mechanism in relation to the protective effect of ginsenoside Rg3 on stressed broilers especially the metabolites alteration remains obscure. The present study aimed to investigate the underlined mechanism in relation to the pathogenesis and protective effect of ginsenoside Rg3 on stressed broilers using liquid chromatograph-mass spectrometry profiling. Eighteen broiler chicks were randomly allocated to 3 treatments: Control, Model and Rg3. Chickens in Rg3 group received intraperitoneally administered 1 mg/kg Rg3 2 h before LPS challenge. Then the broilers were intraperitoneally injection of 250 µg/kg LPS at the age of 12, 14, 33, and 35 d to induce immune stress. Control group was injected with an equivalent amount of sterile saline. At the end of the experiment, the serum was obtained for metabolomics analysis. The changes in serum metabolic profiles were investigated with the application of metabolomics approach. Distinct changes in metabolite patterns in serum were observed by orthogonal partial least square-discriminate analysis. In total, 35 metabolites were identified, among which 17 differential metabolites were found between Control and Model group, and 18 differential metabolites were identified between Model and Rg3 group. Metabolic pathway analysis revealed potential serum metabolites involved in oxidative stress and inflammation, degradation of lipid and protein in broiler chicks with immune stress. In addition, the protective effect of Rg3 on the stressed chicks may be largely mediated by BCAA metabolism, apoptosis and mTOR signaling pathway. These results suggested the potential biomarkers involved in pathogenesis and prevention of stress induced by Escherichia coli lipopolysaccharide.

Ginsenoside Rg3 Ameliorates Stress of Broiler Chicks Induced by Escherichia coli Lipopolysaccharide.

In broiler chicks, Escherichia coli lipopolysaccharide is a prominent cause for inflammatory damage and loss of immune homeostasis in broiler chicks. Ginsenosides have been shown to have anti-inflammatory and antioxidant effects. However, it has not been demonstrated that ginsenosides protect broiler chicks against stress induced by Escherichia coli lipopolysaccharide challenge. The aim of this is to investigate the protective effect of ginsenosides Rg1, Re, and Rg3 on Escherichia coli lipopolysaccharide-induced stress. Our results showed that Rg3 ameliorated growth inhibition and fever, as well as decreased the production of stress-related hormones in broilers with stress. The protective effect of Rg3 on the stressed chicks may be largely mediated by regulating inflammatory response and oxidative damage. Moreover, real-time quantitative-polymerase chain reaction (RT-qPCR) results demonstrated that Rg3 upregulated mRNA expression of mTOR, HO-1, and SOD-1. These results suggested that ginsenoside Rg3 and ginsenoside products contains Rg3 deserve further study for the control of immunological stress and inflammation in broiler chicks.