Effect Of High Concentrate Diet On Apoptosis In The Liver Of Ruminants And The Regulation Of Sodium Butyrate

Lipopolysaccharide(LPS)is a major component of Gram-negative bacteria cell wall,and the main factor to induce the inflammatory damage of the body.In modern animal husbandry,in order to pursue high yield of meat,dairy products,and obtain a greater profit,the animals were fed high grain proportion diet to meet the needs of their own high yield performance.Excessive easy fermentation concentrate lead to lower rumen acidity,gram negative bacteria die and release LPS,causing inflammation of the body.However,liver is a crucial immune organ,which is known as the processing factory of the body,involved in digestion and absorption,protein synthesis,detoxification of toxic substances and immune response.When excessive LPS are accumulated in the liver,the liver is vulnerable to LPS,so that the structure of liver changes,the function is damaged and the degree of apoptosis increased.This experiment was divided into three parts to research the effect of high concentrate diet on apoptosis in the liver of ruminants and the regulation of sodium butyrate.1.Effect of high concentrate diets on hepatic cell apoptosis in dairy cows.Twelve mid-lactating Holstein cows with rumen fistula and hepatic,portal vein cannula were used to investigate the effect of high concentrate diet on hepatic cell apoptosis in ruminants.The experiment was randomly divided into two groups:High-concentrate diet group(HC)and Low-concentrate diet group(LC).Rumen pH value was measured via collecting 0-12 h rumen fluid after feeding.The contents of GOT,GPT,LDH,ALB and T-Pro in peripheral blood were detected by biochemical analysis method to detect the damage degree of the liver of dairy cows under the condition of high concentrate diet.The content of LPS in the liver was detected via the portal vein and hepatic vein blood.The expression of genes was detected by RT-qPCR.The concentration of inflammatory factors IL-1,IL-6 and TNF-α were detected by radioimmunoassay.The activity of Caspase-3 and-8 were determined by spectrophotometer and the protein expression of NF-κB was measured by Westernblot.The results showed that the rumen pH in HC group was below 5.6 and continued for more than 3 h per day after feeding 20 weeks,showing the occurrence of SARA in dairy cows.In HC group,the concentration of LPS in the portal vein was significantly increased,and the liver function was also affected,the pro-apoptotic genes were up-regulated and anti-apoptosis gene was down-regulated,the concentrations of inflammatory cytokines were significantly increased,NF-κB protein expression increased,Caspase-3.8 enzyme activity were up-regulated and caspase-3 was significant differences controlled with LC group.In conclusion,the LPS in digestive tract were transferred into liver via portal vein,causing liver inflammatory damage and increasing the degree of hepatic cells apoptosis2.Effect of high concentrate diets on the apoptosis of liver cells in dairy goats and the regulation of sodium butyrate.Long-term high concentrate diets could induce the occurrence of SARA in ruminants,and LPS coming from the cleavage of gram-negative bacteria in the rumen transfered to liver via portal vein,resulting in liver damage and apoptosis.In this experiment,18 mid-lactation Saanen diary goats were fed with low-concentrate diet,high-concentrate diet and adding sodium butyrate in high-concentrate diet,in order to study the effect of high concentrate diets and sodium butyrate on the apoptosis of liver cells in dairy goats.The 18 dairy goats were randomly divided into three groups:low-concentrate group(LC),high-concentrate group(HC),sodium butyrate + high-concentrate group(SHC).After feeding 20-week,collecting 0-10 h rumen fluid by rumen fistula,and detected the pH value.The liver function was detected by biochemical analysis to determine the degree of liver damage.The content of LPS in the liver was detected via the portal vein and hepatic vein blood.The expression of genes was detected by RT-qPCR.The concentration of inflammatory factors IL-1,IL-6 and TNF-α were detected by radioimmunoassay.The results showed that after feeding 20 w,the rumen pH in HC group was below 5.8 and continued 4 h,this indicted that dairy goats occurred SARA,but in SHC group the pH was between LC and HC group,reducing the continuous decrease in pH.Compared with LC and SHC group,the serum enzyme activity were up-regulated,protein content was down-rugulated,liver function was damaged to some extent,the concentration of LPS in portal vein,hepatic vein was significantly increased,pro-apoptosis genes were up-regulated and anti-apoptosis gene was down-regulated,and the concentration of TNF-α,IL-1β and IL-6 was increased significantly in peripheral blood.In conclusion,long-term feeding high concentrate diet could increase the LPS concent in liver that came from digestive tract,and cause inflammatory liver injury,increase the degree of liver apoptosis,and sodium butyrate could alleviate the liver function damage and the degree of apoptosis.3.Study on the signal pathway of the liver apoptosis in dairy goats with high concentrate diets and the regulation of sodium butyrate.Apoptosis is a spontaneous death process after pathological or physiological stimulation,which is controlled by genes and a series of enzymes.Apoptosis is divided into intrinsic and the extrinsic apoptotic pathways.In order to study the effect of high concentrate diets and sodium butyrate on the apoptosis of liver cells and the main signal pathway in the apoptosis process,18 mid-lactation Saanen diary goats were divided into three groups:LC,HC,SHC.The protein expression of TNF-α,NF-κB(p65,Phospho-p65),cytochrome C,Bcl-2,Bax,Caspase-3 and activated-Caspase-3 were determinated by westernblot.The enzyme activities of Caspase-3 and Caspase-8 in liver tissue were measured by Spectrophotometry.The results showed that,compared with the LC group,the protein expression of TNF-α,NF-κB(p65,Phospho-p65),cytochrome C,Bax,Caspase-3 and activated-Caspase-3 and the enzyme activity of Caspase-3,-8 were all increased significantly,the Bcl-2 protein was reduced significantly in HC group.Compared with the HC group,the protein expression of TNF-α,NF-κB(p65,Phospho-p65),cytochrome C,Bax,Caspase-3 and activated-Caspase-3 and the enzyme activity of Caspase-3,-8 were all down-regulated,the Bcl-2 protein was up-regulated.Therefore,during high concentrate diet induced liver apoptosis and the regulation of sodium butyrate on apoptosis,the two apoptotic pathways were all involved.