Mechanism Research On The Treatment Of Emodin For Severe Acute Pancreatitis

ObjectiveTo investigate the influence of emodin on the severity of the disease and every organ pathology in rats with severe acute pancreatitis, confirming its role in the treatment of severe acute pancreatitis; To explan the mechanism of the treatment of emodin for severe acute pancreatitis by analysing the effect of emodin on the cytokine levels, Treg proportion and function.MethodsSixty Wistar rats were randomly divided into three groups(n=20):(1) the sham-operated control group;(2) the severe acute pancreatitis(SAP) group;(3) the emodin-treated group. The severe acute pancreatitis model was established by using intrapancreatobiliary duct injection of 5% sodium taurocholate. the emodin-treated group:the 2.5mg/Kg emodin was slowly injected immediately after the establishment of SAP model, The same amount of saline solution was injected in other two groups. The rats were killed after 24 hours, the serum amylase, lipase, AST, ALT levels were analysised with automatic biochemical analyzer and the MPO vitality of Supernatant of lung homogenates with Spectrophotometer, calculate the ratio of lung wet to dry weight. Detecting the pathological damage of pancreatic, lung and liver with HE staining methods. The serum TNF-α, IL-6, IL-10, MCP-1 levels were detected with enzyme-linked immunosorbent assay(ELISA). CD4+ and Foxp3+ T lymphocytes were labeled by using direct immunofluorescence assays, detecting the percentage of each group Treg and Treg/CD4+ ratio by flow cytometry. The detecting of Foxp3 and CTLA-4 m RNA expression were measured by Real-time PCR.Results1. The serum lipase, serum amylase, AST, ALT, lung MPO activity and lung wet to dry weight ratio in SAP group were significantly higher than those in sham group(P<0.05), the pancreas,liver, lung pathological damage in SAP group were obvious and the histological scores were higher than those in sham group(P<0.05); The serum indicators and MPO activity in emodin-treated group were decreased comparing with the SAP group(P<0.05), while the organs pathological damage was mitigation and histology scores were decreased(P<0.05).2. The serum IL-6, TNF-α, MCP-1 levels in SAP group were higher than those in sham group(P<0.05), but IL-10 was lower(P<0.05); the serum IL-6, TNF-α, MCP-1 levels in emodin-treated group were decreased significantly comparing with the SAP group(P<0.05), but the IL-10 level was slightly increased(P>0.05).3. The proportion of Treg and Treg/CD4+ T cell ratio were n SAP group were higher than those in sham group(P<0.05), while the expression of Foxp3 and CTLA-4 m RNA were enhanced(P<0.05); the proportion of Treg and Treg/CD4+ T cell ratio in emodin-treated group were decreased comparing with the SAP group(P<0.05),while the expression of Foxp3 and CTLA-4 m RNA were declined(P<0.05).Conclusion1. The model of SAP with intrapancreatobiliary duct injection of 5% sodium taurocholate, test confirmed: the pancreas, lung and liver had be found varying degrees of edema, hemorrhage, necrosis and inflammatory cell infiltration, the model was relatively stable and repeatability.2. Emodin can reduce the severity of severe acute pancreatitis rat and the Pathological damage of every organ.3. Emodin can adjust the balance of Pro-inflammatory anti-inflammatory in peripheral blood of severe acute pancreatitis rat, At the same time can reduce the proportion of Treg and the function of Treg activity, improve immune suppression, so as to achieve the effect of the treatment of severe acute pancreatitis.