The Effect Of Daisaikoto's Treatment To Acute Pancreatitis On Mitochondria Protein Of Pancreatic Acinar Cell's

1.Research backgroundAcute pancreatitis(AP)is a common severe disease in clinics.It can be divided into two types that are mild acute pancreatitis(MAP)and severe acute pancreatitis(SAP).The SAP often compound with severe complications,shock would occur in the early stage,and leads to MODS,in sever cases,MOF will be happening. 100 years had passed,AP passed the "operation-conservation-early operation-individual integrated treatment".Now early operation has beengiven up,conservation has been dominantly,and the changing of the disease has been observed carefully.If any bad condition happened,the operation would need at once.Until now the etiology and pathogenesis of AP are still obscure,and are thoughtas multiple factors.There are still no specific and effective treatment for AP,the death rate is still high.The principle of the treatment are decreasing the pancreas juice secretion,inhibiting the enzyme activity of pancreas,preventing complications.The operation indications of SAP are necrosis with inflammation of pancreasgland.There is no special term for AP in TCM,but in TCM history,there were many descriptions about this disease.Daisaikoto soup is the basic formula in treating AP,and result is verygood.Clinical experiment shows that after using Daisaikoto Soup,the local and systemic symptoms and signs were relieved quickly.Animal experiment shows that it can inhibit the secretion of the trypsin,pancreatic lipase and the other enzymes refereed to pancreas,benefit drainages of bile duct,decrease the tension of Oddi's sphincter muscle,change blood irrigation of thegastrointestinal mucosal layer,relive tissue and organ ischemia and hypoxia,benefit for the recovery of thegastrointestinal mucosal layer, inhibit the over-activity of macrophage and the other immune functional cells, decrease the production of the inflammation cell factors and other inflammation medium.Proteomics is an subject which investigate for the compound and rules of the whole proteins in the cell.It provides the theoretical basis of revealing vital movements,probing the mechanical of those severe disease,diagnosis and prevention of the disease,and the creating the new drugs.The 2-DE and MS are the main supporting techniques.2-DE is the most important and widely used protein separating method in proteomics research,and is the highest resolving method.It is concernedgreatly in recent years.MS is a way which through purifying intracellular substances and protein compounds,identifying protein components and it's subcellualr position.It is suitable for those micro-organ samples with complicated components.Mitochondrial is one of the most important and complicated intracellular organs.It is in the matrix of aerobic eukaryocyte.It is the energy supplying centre in the intracellular matrix.The functional changes of the mitochondrial would refer to the cell fading,degeneration and tumor cells. It is the hot point of clinical and basis research.2.Research purposeDaisaikoto soup is the main formulae in treating AP nowadays.But it's researching still on clinical observation and hormone level.So using AP animal model as researching project,using immunology and proteomics methods,we studies the injury of mitochondrial of AP granular cells.We hope to find a new theory and new thinking in proteomics researching of treating AP with TCM.3.Research plans3.1.Clinical research 36MAP confirmed cases were divided into western medicine combined placate treatmentgroup and western medicine combined with Daisaikoto Soup treatmentgroup randomized.The duration of treatment was 7 days.The check index was observation the change of serum amylase of MAP patients.3.2 Animal research3.2.1 Making AP model:32 SD mice were divided into contrastgroup(15)and experimentgroup(15)randomized to be making AP model,the remain 2 were normal blankgroup.3.2.2 Prescription 12 hours after operation the mice of contrastgroup weregiven NS 5ml/24hr orally.The mice of experimentgroup weregiven Daisaikoto Soup 5ml/24hr orally the same.3.2.3 Collection of sample:The mice of contrast and experimentgroup were killed after 12,24,72 hours whichgiven different treatment.5 mice were killed each time.The killed mice were taken off the pancreas(5g)to be made pancreatic acinar cells.3.3 Experiment procedure of mice pancreatic acinar mitochondria:The pancreas of killed mice were taken of immediately and disposed to collect pancreatic acinar cells.The pick-up procedure was using mitochondria and plasma protein boxes.The mitochondria were placed -80℃.3.4 The proteins of mitochondria were analyses by 2-DE and MS technology.4.ResultThere is agreat difference of patient's serum amylase between western medicine combined with Daisaikoto Soup treatmentgroup and western medicine combined placate treatmentgroup(p＜0.05).2-DE research shows that through the experiment of proteomics expression in AP mice model,the expression of mitochondria protein of pancreatic acinar cells hadgreat difference with normal condition.After Daisaikoto Soup treatment,the losing and specific protein expression had some degree recovery, and presenting a series of new proteins which had some difference with the normal proteins.Using MS technology,we find some specific and difference protein expression in mitochondria protein of pancreatic acinar cells with Daisaikoto Soup treatment.5.Conclusion2-DE research shows that there are series of proteins expression turbulence in AP condition,such as changing of protein expression,lack of protein expression,and specific protein expression.Those problems are accordant with TCM principle of integration concept and treatment according to clinical manifestation.After treating the AP mice with Daisaikoto Soup, the problems which lacking protein expression and specific abnormal expression have recover in some extent.Some changes that differ from normal conditions have happened.It shows that difference protein must have some relations with the happening and treating of AP on AP models.Daisaikoto Soup must make some effect in the regulation of correlation proteins.The treating function of Daisaikoto Soup have changed the expression and modifying of those specific protein perhaps.Using MS technology we find some expression of specific proteins.Though due to restricted by some poor conditions,we still can not illustrate the function of these proteins very clearly.What's the relationship between them with the mechanism of AP? Bow Daisaikoto Soup does treat AP through affect the expression of those proteins? But those difference proteins will provide some important message in treating AP,and build agood base in discuss the function of Daisaikoto Soup in treating AP in molecular level and clinically.