The Research On Regulating Of Pancreatic Stellate Cells Activation And Signal Pathway Correlated With Pancreatic Fibrosis Using The Method Of Invigorating Spleen To Resolve Dampness And Dredging Collaterals

In traditional Chinese medicine (TCM) theory, the spleen and stomach are the source of qi and blood, the pivot of functional activities of qi’s going up and down. Briefly speaking,"Spleen stomach is Pivot". Only when the ascending and descending functions of spleen and stomach is regular can the movement of qi and blood be in harmony, which is adjustment mechanism of internal balance and is central part of clarifing human physiological activity and the pathological mechanism and is also important for syndrome differentiation and treatment. According to the TCM theory, the pancreas is part of spleen-stomach which is a multifunctional composite unit including the digestive system and other systems. Many miscellaneous diseases due to internal injury can be treated from spleen and stomach in the TCM. Pancreatic fibrosis is one of the most frequently-occurring disease of digestive system, and it has not made any breakthrough progress in it’s treatment at present. Hence interpretating the occurrence and development and treatment of Pancreatic fibrosis basing on Spleen-Stomach theory in TCM has important academic and clinical value. Under guidance of Spleen-Stomach theory in TCM, this article try to explain Pancreas fibrosis’pathogenesis and treatment rules from two aspects-principle in TCM and cellular level.In theoretical research: Firsrly, we can attribute Pancreas fibrosis to Pi diseases in TCM because of the anatomy of Pancreas and its function and Clinical common symptom of Pancreas fibrosis. Secondly, from the aspects of pathogenesis, the article thinks that the dampness blocking collaterals of spleen due to dysfunction of spleen in transforming and transporting is the key pathogenesis of Pancreas fibrosis. In view of its characteristic, the method of nourishing spleen and resolving dampness is the important treatment for pancreatic fibrosis, and the method of dredging collaterals is important compatibility in its treatment.In the experimental aspect: in view of the activation of pancreatic stellate cells (PSCs) is the key point to the formation of pancreatic fibrosis, we take activated PSCs as the research object to investigate the impact of invigorating spleen to resolve dampness and dredging collaterals on cellular activation and proliferation of cultured rat PSCs and to explore its mechanisms using Modern Biological Technology. Fistly, we carry out isolation, culture, and identification of rat PSCs using tissue mass method. The morphology of cell, fat droplets and skeleton protein fully prove that cell in vitro model for Pancreatic fibrosis was successfully constructed. Secondly, we screened the effect of the drug serum through the method of serum pharmacology and MTT assay, and ascertain the best medical concentration of the drug serum. The results show that the method of invigorating spleen to resolve dampness and dredging collaterals can restrain the activation and proliferation of rat PSCs and suppress Pancreatic Fibrosis, and the optimal concentration is10%. Because of the impotant effect of TGF-β1signal transduction pathway on the activation and proliferation of rat PSCs, we analysed the cell supernatant by the method of enzyme-linked immunosorbent assay (ELISA), the results show that the method of invigorating spleen to resolve dampness and dredging collaterals restrain the secretion and expression of TGF-β1in rat PSCs, therefore affecting activation and proliferation of rat PSCs by regulating and controlling of TGF-β1signaling transduction pathway is one of its anti-fibrotic ways.