The present study was conducted in vitro and in vivo. The first in vitro study was performed to describe the effect of different exposure duration of 37°C and 41°C temperature on cell viability, apoptosis, cell cycle, and heat shock protein expression in broiler satellite cells and fibroblast isolat...
The present study was conducted in vitro and in vivo. The first in vitro study was performed to describe the effect of different exposure duration of 37°C and 41°C temperature on cell viability, apoptosis, cell cycle, and heat shock protein expression in broiler satellite cells and fibroblast isolated from breast muscle of one-day-old broiler chicks. The second in vivo study was performed to describe the effect of different duration of acute and chronic heat stress on growth performance, liver enzyme, intestinal histology and expression of heat shock proteins and different cytokines of proteins and gene.
The first experiment inspects the effects of heat stress at different heat exposure time on satellite cells of broiler. The broiler satellite cells isolated from breast muscle of one-day-old chicks for this study. This satellite cells treated by 37°C and 41°C temperature for 6 h, 12 h, 24 h, 48 h, and 72 h. Interestingly, cell viability significantly higher viable cells were found at 24 h and 48 h in both temperatures. At 12 h, remarkably higher number of viable satellite cells found at 41°C compared to 37°C temperature. Besides, higher percent of live cells were found at early duration of 41°C exposure compared to 37°C temperature. However, significantly higher percentage of apoptotic cells was found at 72 h in 41°C than 37°C. The protein and gene expression of heat shock protein (HSP) 70, HSP60, and HSP47 were remarkably increase during the experimental period at temperature of 41°C than 37°C. The results of this study suggest that the broiler satellite cells viability and heat stress related proteins or genes were highly expressed at 41°C temperature where at 37°C lower expression of proteins and genes of HSPs. Eventually, this study suggests that 41°C can accelerate the broiler satellite cell viability compared to 37°C at early incubation time.
In the second experiment, there is increasing interest in specific temperatures to increase the efficiency of animal production. Interestingly, fibroblasts, connective tissue cells that produce collagen, are essential for repairing tissue damage and can be effective in animal production. The aim of the current study was to determine the mild heat stress temperature at which broiler fibroblast viability would be highest for the longest period. The stress level was measured as the expression level of heat shock proteins (HSPs). The full sample was divided into two groups, control or experimental, and each of these was divided into five subgroups based on incubation time with three replications. Broiler fibroblast viability and HSP expression levels were measured according to incubation times and temperature. At the experimental temperature, fibroblast viability increased significantly at 12 h but decreased at 72 h incubation time compared with the viability in the control group, whereas with 24 h and 48 h incubation time, the viability rates increased significantly at both temperatures. Live cell viability at the experimental temperature increased significantly at 24 h and then significantly declined at 48 h and 72 h, in contrast with viability at the control temperature. However, higher percentages of fibroblasts were found with 24 h and 48 h incubation time at the experiment and control temperatures, respectively. Moreover, the S phase lengthened significantly with 6 h, 12 h, and 24h incubation time in the experimental group but not in the control group. Protein and mRNA (HSP70, HSP60, and HSP47) expression were significantly higher in the experimental group than in the contr-ol group, but at the end of the experimental period, HSP expression level was high in both temperature groups. At the experimental temperature, higher viable fibroblasts were found in shorter incubation times, although longer exposure at this temperature decreased the cell viability owing to increased apoptosis. HSP expression, however, did increase at this temperature.
The third experiment inspects the effect of time based different acute heat stress on liver specific enzymes (SGPT and SGOT) and HSPs protein and gene expression of broiler. A total of three hundred broilers allocated to control and treatment group for this experiment. The treatment group was divided into 4 subgroups such as 3, 6, 12, and 24 h based on acute heat exposure duration. In this experiment was determined the protein and gene expression of HSP70, HSP60, and HSP47 of 3 different sections of broiler small intestine during exposure of acute heat stress. In duodenum and jejunum, the HSP70 and HSP60 expression of the protein was significantly higher at 6 h acute heat exposure, but the expressions of the protein of HSP70 and HSP60 were significantly higher at 12 h heat exposure in the ileum. In duodenum and jejunum, the protein expression of HSP47 was remarkably higher at 3 h heat exposure, in the ileum, this protein showed significantly higher expression at 6 h heat exposure. The gene expression level of HSP70, HSP60, and HSP47 were significantly higher due to acute heat stress at different time points compared to control in three different sections of the broiler's small intestine. The SGPT and SGOT levels were significantly higher at 12-h and 24-h respectively than other heat exposure duration. The result of this study confirmed that acute heat stress affects intestinal proteins and gene expression of broiler until inducing heat tolerance.
In the final experiment of this study, we inspected the small intestinal HSPs expression and the relationship between HSPs and the immunity of broiler during chronic heat stress. This study was divided into control and heat stress groups with eight replications per group. A total of 15 broilers were assigned for each replication. Furthermore, on day 35, the treatment group was subdivided into heavy and low body weight groups based on the body weight. On days 28 and 35, Due to chronic heat stress the growth performance of broiler reduced than control group, and the protein expression of HSP70, HSP60, and HSP47 increased in the heat stress group compared with the control group. The protein and gene expression level of HSPs were remarkably higher in the low body weight group than in the control group. The gene expression of HSP70 and HSP60 were significantly co-related to pro- and anti-inflammatory cytokines in the broiler small intestine in the treatment group. Therefore, the HSP70 and HSP60 activated the adaptive immunity in the broiler small intestine because of exposure to chronic heat stress. Eventually, it may increase heat tolerance capacity for the broiler.
The present study was conducted in vitro and in vivo. The first in vitro study was performed to describe the effect of different exposure duration of 37°C and 41°C temperature on cell viability, apoptosis, cell cycle, and heat shock protein expression in broiler satellite cells and fibroblast isolated from breast muscle of one-day-old broiler chicks. The second in vivo study was performed to describe the effect of different duration of acute and chronic heat stress on growth performance, liver enzyme, intestinal histology and expression of heat shock proteins and different cytokines of proteins and gene.
The first experiment inspects the effects of heat stress at different heat exposure time on satellite cells of broiler. The broiler satellite cells isolated from breast muscle of one-day-old chicks for this study. This satellite cells treated by 37°C and 41°C temperature for 6 h, 12 h, 24 h, 48 h, and 72 h. Interestingly, cell viability significantly higher viable cells were found at 24 h and 48 h in both temperatures. At 12 h, remarkably higher number of viable satellite cells found at 41°C compared to 37°C temperature. Besides, higher percent of live cells were found at early duration of 41°C exposure compared to 37°C temperature. However, significantly higher percentage of apoptotic cells was found at 72 h in 41°C than 37°C. The protein and gene expression of heat shock protein (HSP) 70, HSP60, and HSP47 were remarkably increase during the experimental period at temperature of 41°C than 37°C. The results of this study suggest that the broiler satellite cells viability and heat stress related proteins or genes were highly expressed at 41°C temperature where at 37°C lower expression of proteins and genes of HSPs. Eventually, this study suggests that 41°C can accelerate the broiler satellite cell viability compared to 37°C at early incubation time.
In the second experiment, there is increasing interest in specific temperatures to increase the efficiency of animal production. Interestingly, fibroblasts, connective tissue cells that produce collagen, are essential for repairing tissue damage and can be effective in animal production. The aim of the current study was to determine the mild heat stress temperature at which broiler fibroblast viability would be highest for the longest period. The stress level was measured as the expression level of heat shock proteins (HSPs). The full sample was divided into two groups, control or experimental, and each of these was divided into five subgroups based on incubation time with three replications. Broiler fibroblast viability and HSP expression levels were measured according to incubation times and temperature. At the experimental temperature, fibroblast viability increased significantly at 12 h but decreased at 72 h incubation time compared with the viability in the control group, whereas with 24 h and 48 h incubation time, the viability rates increased significantly at both temperatures. Live cell viability at the experimental temperature increased significantly at 24 h and then significantly declined at 48 h and 72 h, in contrast with viability at the control temperature. However, higher percentages of fibroblasts were found with 24 h and 48 h incubation time at the experiment and control temperatures, respectively. Moreover, the S phase lengthened significantly with 6 h, 12 h, and 24h incubation time in the experimental group but not in the control group. Protein and mRNA (HSP70, HSP60, and HSP47) expression were significantly higher in the experimental group than in the contr-ol group, but at the end of the experimental period, HSP expression level was high in both temperature groups. At the experimental temperature, higher viable fibroblasts were found in shorter incubation times, although longer exposure at this temperature decreased the cell viability owing to increased apoptosis. HSP expression, however, did increase at this temperature.
The third experiment inspects the effect of time based different acute heat stress on liver specific enzymes (SGPT and SGOT) and HSPs protein and gene expression of broiler. A total of three hundred broilers allocated to control and treatment group for this experiment. The treatment group was divided into 4 subgroups such as 3, 6, 12, and 24 h based on acute heat exposure duration. In this experiment was determined the protein and gene expression of HSP70, HSP60, and HSP47 of 3 different sections of broiler small intestine during exposure of acute heat stress. In duodenum and jejunum, the HSP70 and HSP60 expression of the protein was significantly higher at 6 h acute heat exposure, but the expressions of the protein of HSP70 and HSP60 were significantly higher at 12 h heat exposure in the ileum. In duodenum and jejunum, the protein expression of HSP47 was remarkably higher at 3 h heat exposure, in the ileum, this protein showed significantly higher expression at 6 h heat exposure. The gene expression level of HSP70, HSP60, and HSP47 were significantly higher due to acute heat stress at different time points compared to control in three different sections of the broiler's small intestine. The SGPT and SGOT levels were significantly higher at 12-h and 24-h respectively than other heat exposure duration. The result of this study confirmed that acute heat stress affects intestinal proteins and gene expression of broiler until inducing heat tolerance.
In the final experiment of this study, we inspected the small intestinal HSPs expression and the relationship between HSPs and the immunity of broiler during chronic heat stress. This study was divided into control and heat stress groups with eight replications per group. A total of 15 broilers were assigned for each replication. Furthermore, on day 35, the treatment group was subdivided into heavy and low body weight groups based on the body weight. On days 28 and 35, Due to chronic heat stress the growth performance of broiler reduced than control group, and the protein expression of HSP70, HSP60, and HSP47 increased in the heat stress group compared with the control group. The protein and gene expression level of HSPs were remarkably higher in the low body weight group than in the control group. The gene expression of HSP70 and HSP60 were significantly co-related to pro- and anti-inflammatory cytokines in the broiler small intestine in the treatment group. Therefore, the HSP70 and HSP60 activated the adaptive immunity in the broiler small intestine because of exposure to chronic heat stress. Eventually, it may increase heat tolerance capacity for the broiler.
주제어
#Heat stress Broiler Satellite cells Fibroblast heat shock protein acute heat stress chronic heat stress 열 스트레스 육계 위성 세포 섬유아세포 열충격 단백질 급성 열 스트레스 만성 열 스트레스
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