The Protective Effect Of Trichostatin A Against Inflammation On Acute Liver Failure Rats

Part Ⅰ The effects of histone deacetylase 1 siRNA on lipopolisaccharide-induced RAW264.7 cellsObjective:When the immunity of human body was declined,the lipopolisaccharide from primary nidus or damaged intestine will be released into blood flow,causing endotoxemia.Usually,endotoxemia will activate various immune cells to trigger immunoreation,such as macrophages,subsequently induce multi-organ disorder or failure.To observe the effects of histone deacetylase(HDAC)1 silencing on lipopolysaccharide(LPS)induced inflammation,the present study applied HDAC1 siRNA to treat the LPS-induced RAW264.7 cells.Methods:At first,we optimized the best sequence among four HDAC1 siRNA by detecting the HDAC1 protein expression.RAW264.7 cells were seeded in 6-well plates,and divided into 3 groups,control group,LPS group and siRNA group.When the cell fusion reached to 40%,HDAC1 siRNA was introduced into the cells medium at a final concentration of 50nM.After a 4-hour transfection,50ng/ml LPS were added to both LPS group and siRNA group.After 48 hours,the cell morphology and proliferation were observed.The changes of total HDAC activity in culture medium and cells’ inflammatory factors were detected.Results:HDAC1 siRNA inhibited the differentiation and proliferation of LPS-induced RAW264.7 cells.Not only HDAC1,but HDAC3 and HDAC4 were down-regulated by HDAC1 siRNA.While compared with control group,the expression of inflammatory factors was increased in LPS group and subsequently decreased in siRNA groups.Conclusion:HDAC1 silencing confered protective effect to LPS-induced RAW264.7 cells through inflammation inhibition.Part Ⅱ The heptic protective effects of trichostatin A on acute liver failure ratsObjective:Acute liver failure is a severe clinical disease with a significant feature of massive necrosis of hepatic cells.Due to severe liver function damage,acute liver failure usually results in multi-organ dysfunction or failure.Using trichostatin A(TSA)to treat acute liver failure rats,the study aimed to evaluate the effects of TSA on the liver of acute liver failure rats.Methods:The acute liver failure model were induced by D-galactosamine and LPS.The SD rats were randomly divided into control group,model group and TSA group.Two hours prior to model creation,TSA was administrated at a dose of 200 mg/kg,while the control group and model group were treated with normal saline at the same amount.The expression of HDACs,levels of histone acetylation and inflammatory factors,and changes of liver function and histology were detected after a 48-hour LPS stimulation.Results:Compared with model group,TSA significantly inhibited the expression of HDACs,promoted histone acetylation,improved the liver function and histology,and down-regulated the expression of inflammatory factors as well.Conclusion:TSA has protective effects on the liver of acute liver failure rats via inhibiting inflammation.Part Ⅲ The protective effects of trichostatin A on the intestine of acute liver failure ratsObjective:The "two-hit" hypothesis is one of the main mechanisms in acute liver failure.Intestinal endotoxin plays an important role in the process of acute liver failure.Usually,the liver dysfunction causes damage to intestine,then LPS passes through the intestinal wall and goes into blood flow.The endotoxemia subsequently harms to liver function,forming a vicious circle.Using TSA to treat acute liver failure rates,this study aimed to evaluate the effects of TSA on the intestine of acute liver failure rats.Methods:The acute liver failure model were conducted by D-galactosamine and LPS.Eighteen SD rats were randomly divided into 3 groups,control group,model group and TSA group,6 for each group.Two hours prior to the models creation,TSA was administrated at a dose of 200 mg/kg,control group and model group were treated with normal saline as control.Forty-eight hours after LPS stimulation,the expression of HD AC,levels of histone acetylation and inflammatory factors,changes of intestinal permeability and histology of small intestine were detected.Whole colon was taken out for motility observation.Results:Compared with model group,TSA significantly inhibited the expression of HDACs,promoted levels of histone acetylation,repaired the damage to intestinal permeability and histology,and down-regulated the expression of inflammatory factors as well.The colonic motility disorder in model group was corrected by TSA.Conclusion:TSA has intestinal protection on the acute liver failure rats through inflammation inhibition.