Shibuya Intestinal Diarrhea. Nudicaule And Owned By The Large Intestine By In Vivo Experiments

Objectives:Based on the Papaver nudicaule L. and its total alkaloids have the effect of anti-diarrhea type ulcerative colitis in the clinical and pharmacological research and work foundation. This study will be on the basis of that, mainly reference in Chinese traditional medicine theory and application, using chemical separation and analysis technology, combined with the modern molecular biology experiment method, from the point of view on active ingredients of Papaver nudicaule L. distributed in different organs or tissues in the rat and influence on its intracellular of the second messenger cAMP, cGMP content, provides experimental evidence to Chinese meridian theory of Papaver nudicaule L.. At the same time trying to analysis content of VIP and expression of VIPRmRNA in colon tissue of rats, reveal the mechanism of Papaver nudicaule L. function on anti-diarrheal.Methods:1. Using the method of senna with acetic acid stimulation, replication the model of UC by rat;2. Using ethanol extract and weak base precipitated, chloroform extraction TAPN;3. Using the method of HE staining and observe the pathological changes in colonic tissue of rats by light microscopy;4. Using the method of HPLC-MS, qualitative detected the distribution of nudicauline in different visceral organ at different time points;5. Using the method of ELISA, detected the content of AC, cAMP, cGMP in different visceral organ;6. Using the method of ELISA and real-time RT-PCR, detected the content of VIP and expression of VIPRmRNA in colon tissue of rats.Results:1. Compared with Model group, diarrhea index decreased significantly in PN group and Loperamide group (P<0.01), the extent of occult blood converted to weakly positive, some of them had shown negative; pathological shown that colonic epithelial integrity, arrange evenly, glands lined neat, only a small amount of inflammatory cell infiltration;2. Through the mass spectrometry to visceral organ of PN group rats, nudicauline has been found in stomach and large intestine at PN treatment group of different time points;3. Compared with Model group, content of AC increased significantly in spleen, large intestine, small intestine(P<0.01or P<0.05), decreased significantly in heart(P<0.01) at PN group; content of cAMP increased significantly in stomach(P<0.05), decreased significantly in large intestine(P<0.01) at PN group; content of cGMP increased significantly in large intestine, brain(P<0.01or P<0.05), decreased significantly in lung(P<0.01) at PN group; the ratio of cAMP/cGMP decreased significantly in large intestine(P<0.01) at PN group; between content of AC and levels of cAMP were significantly correlated in liver(P<0.01or P<0.05) at PN group;4. Compared with Model group, content of VIP and expression of VIPRlmRNA decreased significantly in colon tissue (P<0.01) at PN group, expression of VIPR2mRNA also showed downward trend, however, no statistical significant difference(P>0.05).Conclusions:1. TAPN could effectively reduce the diarrhea index and blood in the stool, obviously improve the symptoms of diarrhea rats;2. TAPN could promote damage colon tissue repaired, restore colon tissue's function in diarrhea rats;3. After the effects of TAPN, nudicauline could be detected at stomach and large intestine in diarrhea rats;4. TAPN could change the content of cyclic nucleotide or the ratio of it obviously in the diarrhea rats' stomach, lung, large intestine, brain;5. TAPN could decrease the content of VIP and reduce the expression of VIPRlmRNA in diarrhea rats'colon tissue.6. From the distribution of Nudicauline in rat's large intestine, change of VIPRmRNA expression in colon tissue and content of cAMP and cGMP in intracellular of different organs, offer a part of the experiment basis to clarification of the theory Papaver nudicaule L. owned by the large intestine.