| Objective Severe acute pancreatitis is characterized by a poor prognosis with local and systemic complications, high morbidity and mortality. The clinical presentation of acute pancreatitis varies significantly from mild self-limiting discomfort to a severe life-threatening condition. Once the disease process is initiated, the severity of the disease is largely determined by a complex network of activated inflammatory mediators such as cytokines, proteolytic enzymes, reactive oxygen species, and many more which render the local injury to a systemic disease with multiple organ dysfunction, sepsis, and considerable mortality. Remarkable progress in diagnostic modalities, intensive care technologies, and organ preserving surgical techniques have decreased mortality of severe acute pancreatitis during the past decades. However, the treatment of acute pancreatitis still remains largely supportive and no specific approach exists to prevent evolving complications. According to the report, the mortality have been decreased from 70-85% to 10% with traditional Chinese medicine and another therapeutic conditions. We found that Chinese medicine can be showed pharmalogical effects with experiment SAP model. However the mechanism of Chinese medicine is not clear. The objective is the study mechanism of the Chinese medicine (Yijin granules). The Emodin (1,3,8-trihydroxy-6-methylanthraqui- none) is an active component in Yijin granules. Since acute pancreatitis is associated with systemic inflammatory response syndrome, we investigate whether emodin attenuate inflammatory response in the rat with severe acute pancreatitis.Methods At first, SAP model was induced by injection of 3% sodium taurocholic acid into the subcapsular of pancreas. Yijin granule was administered by injecting into the intestines. Plasma amylase of 1.5h, 3h, 6h with SAP was measured and histological examination for pancreas was evaluated by light micmscope.In the second part, the pharmalogical effects of Emodin have been tested. SD rats were divided into four groups randomly: normal control group, SAP model without emodin treatment group and SAP model with emodin treatment group. SAP was induced by injection of 3% sodium taurocholic acid into the subcapsular of pancreas. Emodins ( 5μg and 25μg /kg) were administered by injecting into the intestines. Rats were killed at 12, 24 and 48 hours after the operation. Plasm amylase was measured by means of enzymatic assay with spectrophotometer. The plasma level of IL-1β, TNF-αand IL-10 were evaluated by radioimmunoassay at 12, 24 and 48 hours following acute pancreatitis, and immunohistochemistry was used to assay the expression of P-selectin protein. The lung,hepatic and small intestine tissue histology slides were observed after hematoxylin and eosin staining.In the third part, Effect of Emodin on inflammatory response in severe acute pancreatitis rats' lung have been investigated. SAP model was induced by injection of 3% sodium taurocholic acid into the subcapsular of pancreas. Emodin was administered by injecting into the intestines. Plasma amylase of 12h, 24h and 48h with SAP was measured. Expressions of IL-1β, IL-6 and IL-10 mRNA in lung were detected by semi-quantitative RT-PCR. Immunohistochemistry was used to assay the expression of ICE in lung.In the fourth part, gene chip assay was done for different expression spectrum of gene. The total RNA was extracted from rats's lung, the rats divide with normal control group, SAP model without emodin treatment group and SAP model with emodin group. The sample RAN was a mixture with everyone in same group. The RNA sample were checked and showed good quality, the RNA samples have been delivered to the gene chip assay company for analysis.Resoults The serum amylase levels significantly increased in the SAP rats and could be decreased Yijin granules. The histological examination showed the pancreatic edema in 1.5 h, the pancreatic hemonhage and necrosis with in 3.0-6.0h.The plasma amylase decreased significantly in the emodin-treated group in comparison with non-treated group. The plasma level of IL-1β, TNF-αand IL-8 were decreased significantly in the emodin-treated group in comparison with non-treated group. The plasma level of IL-10 were decreased significantly in the emodin-treated group in comparison with non-treated group.Immunohistochemistry indicates that P-selectin expression was low in normal lungs,hepatic and small intestine tissue, while significantly higher in SAP within non-treated group than normal control and its expression declined in emodin-treated group.To study the mechanism of the Emodin, the pathological lesion score of lung, the mRNA expression of pro-inflammatory cytokin IL-1βand IL-6 decreased significantly in the emodin-treated group in comparison with non-treated group. As well as the lever of expression of ICE decreased significantly in the emodin-treated group in comparison with non-treated group. Moreover, we demonstrated for that emodin upregulated the mRNA expression of IL-10.Gene chip assay indicated the total 35 gene were difference between SAP with and without Emodin. The seven gene were up regulation and another twenty-eight gen were down regulationConclusion1 Chinese medicine Yijin granules can reduce the serum amylase levels in SAP model in rat and decrease the lever of inflammation response.2. The Chinese medicine emodin can ameliorate the systemic inflammatory response during sever acute pancreatitis. The mechanisms might be ascribed to its up-regulation expression of IL-1β, TNF-α, IL-6, IL-8, P-selectin and ICE, and down regulation expression of IL-10.3. The gene chip assay demonstrated that the mechanisms of treatment SAP with Emodin were up regulation and down regulation some gene.
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