The Intervention Of Clearing Away Heat And Removing Toxic Substances(CAHRTS), Cooling Blood To Dissipate Blood Stasis (CBDBS) On Expression Of Proteins In Rats With Endotoxemia And Liver Injury

Objective:To investigate the intervention of clearing away heat and removing toxic substances(CAHRTS), cooling blood to dissipate blood stasis (CBDBS) on expression of proteins in rats with endotoxemia and liver injury.Methods:(1) Forty SD rats were randomly divided into four groups:Normal group, Model group, High dose of traditional chinese medicine (TCM) group, Low dose of traditional chinese medicine (TCM) group. (2) Model group:Model group rats were administered with distilled water 2ml by gastric perfusion continue for 7 days,once daily. One hour after the last gastric perfusion, rats were injected with LPS (10mg/Kg) into its abdominal cavity. Except drinking water freely, rats must fast for more than 12 hours before intraperitoneal injection and blood sampling. Six hours after the intraperitoneal injection, rats were injected with 1% pentobarbital sodium (30mg/Kg) into its abdominal cavity. Anesthetic rats were sterilized with alcohol on the skin, opened chest and sampled blood from hearts. All of the blood was stored in 4℃refrigerator. The serum differential protein expression profiling was detected by SELDI-TOF-MS technology; Liver function enzyme index test was detected by automated hematology analyzer.Serum Endotoxin concentrations was measured by Quantitative Chromogenic Tachypleus Amebocyte Lysate.10% formalin fixed liver tissues (1.5cm×1cm×0.5cm), HE staining routine pathological section were used to observe the pathological changes of liver tissues under light microscope. (3) TCM group:Adult dosage of TCM is 1.58g/crude drug/Kg/per day. Therefore, the conventional dosage of rats are 10.20g/crude drug/Kg/per day which could be obtained by the equation of body surface area. High dose of TCM group were administered with 20.40g/Kg by gastric perfusion continue for 7 days,once daily. Low dose of TCM group were administered with 5.10g/Kg by gastric perfusion continue for 7 days,once daily. The rest of the experimental method was the same as the model group's.(4)Normal group:Normal group rats were administered with distilled water 2ml by gastric perfusion continue for 7 days,once daily. One hour after the last gastric perfusion, rats were injected with the same dose of distilled water into its abdominal cavity. The rest of the experimental method was the same as the model group's. Results:(1) Each group's serum Endotoxin level:compared with model group, the serum Endotoxin level was decreased in the high dose of TCM group and low dose of TCM group, the differences were statistical significance (P<0.05). Compared with low dose of TCM group, the serum Endotoxin level was decreased in the high dose of TCM group, the differences were statistical significance (P＜0.05). (2) Liver biochemistry test showed:compared with model group, the levels of serum ALT,AST were decreased in the high dose of TCM group and low dose of TCM group, the differences were statistical significance(P<0.05). Compared with low dose of TCM group, the levels of serum ALI,AST were decreased in the high dose of TCM group. (3) 115 differently expressed protein peaks were found by protein chip, among which 11 differently expressed protein peaks had statistical significance (P<0.05).Protein 4200Da was high expression in TCM group, protein 8984Da and 9005Da was low expression in TCM group. (4) Observed under optical microscope:①Normal group could be seen clean mark of hepatic plates.Liver.cell sarranged orderliness.Cord structure was clear.②Model group:Inflammatory cell,karyopyknosis and a lot of lipid droplets could be observed.Cord structure was not clear.Hepatic sinus was narrow. Liver cell sarranged disorderliness.③High dose of TCM group could be seen relatively clean mark of hepatic plates and a tiny amount of inflammatory cell.Liver cell sarranged relatively orderliness.Cord structure was clear.④Low dose of TCM group could be seen relatively clean mark of hepatic plates,oncotic liver cells,inflammatory celland congestion around the central veins. Conclusions: (1) The TCM of CAHRTS and CBDBS can reduce the serum Endotoxin level in rats with endotoxemia and liver injury. (2) The TCM of CAHRTS and CBDBS can reduce the levels of ALT,AST in rats with endotoxemia and liver injury. (3) The TCM of CAHRTS and CBDBS have certain intervention role to expression of proteins, which have an effect on expression of proteins in rats with endotoxemia and liver injury. The change of protein peaks may be a significant molecular basis which cause the TCM of CAHRTS and CBDBS intervene rats with endotoxemia and liver injury.The serum assuredly has differential expression protein in rats among groups, which may be of use for farther study.