| ObjectsGuided by the theory of the lung and the large intestine being interior-exteriorly related and under the hot environment of researching on the function of intestinal barrier, we put forward "toxin impairing intestine collateral" pathogenesis theory and deem that damp-heat blood stasis toxin accumulating long damage the intestine collateral and then upward attack the lung is the key cause for the relapses of UC and the lung injury. Afterwards, we establish the rat model of UC, study the injury mechanism of the intestinal mucosal barrier and reveal the inherent links between IMB and lung injury in UC, hope to find some new material foundation for the correlation between lung and intestine, provide strong experimental evidence for the pathogenesis theory, interpret the effects and mechanism of herb monomer including Matrine, astragalus polysaccharides (APS) and the two mixed when used to treat UC and lung injury in UC, deepen the theory of the lung and the large intestine being interior-exteriorly related and then present new ideas and methods also effective drug to solve this difficult medical problem.Methods120healthy male Wistar rats were made to UC models by immune complex method, while another30rats left in normal group. When the model were set up, randomly selected10rats in each group to check whether the model were set up successfully. Then the remaining UC model rats were randomly divided into5groups:normal group, model group, western medicine group, Matrine group, APS group and the two mixed group. Each medicine treatment group was given sulfasalazine enreric-coated tablets, Matrine, APS, and the two mixed separately. Normal and model group were fed with equivoluminal pure water. Observe the general condition of the rats everyday. At the end of0th,2nd,4th week, taking blood from the abdominal aorta and the lung and intestine to tested relevant indicators.Results1. The changes of general condition and weight in UC rats and the effects of the herb monomer.Results:Rats in model group live in a poor life state and compared with the normal group, the weight were significantly loss (P<0.05or P<0.01) while the DAI scores rose (P <0.01), and companied with respiratory symptoms.2nd week, compared with the normal group, the DAI scores of rats in Matrine group and the two mixed group was significantly fallen (P<0.05).4th week, compared with the normal group, the DAI scores of rats in the two mixed group was significantly fallen (P<0.05), while the weight of the rats in APS group and the two mixed group was increased nearly to normal.2. The pathomorphological changes in lung and intestinal tissue of UC rats and the effect of the herb monomer. At the end of0th,2nd,4th week, killed the rats, paraffin embedded the lung and intestinal tissue, then sliced, HE stain and observed the morphological changes.Results:Colonic mucosal hyperemia, edema, erosions, ulcers, necrosis and vasculitis.2nd week, there is no obvious change while at the end of4th week the pathomorphology change in some sort. The lung tissue demonstrated inflammatory cell infiltrate, hyperemia, edema and vasculitis all the time. At the end of4th week, pulmonary fibrosis was obviously. Lung injury is a common parenteral change of UC. Herb monomer can reduce the pathomorphological changes of lung and intestine effectively.3. The damage degree of colonic mucosa and the change of intestinal permeability in UC model rats and the effect of the herb monomer.At the end of0th,2nd,4th week, killed the rats, observed the damage degree of colonic mucosa and scoring. ELISA was used to detect the endotoxin (ET) level in serum, while spectrophotometric method was used to determine the enzyme activity for diamine oxidase, test the intestinal permeability whether changed.Results:The colonic mucosa in the UC model rats showed hyperemia, erosion, ulcer, basic structure disappeared and the vascular texture fuzzy.2nd week, there is no obvious change while at the end of4th week the pathomorphology improved in some sort. Compared with normal group, the CDMI of the model group was significantly increased (P<0.01). At the end of0th,2nd,4th week, the level of ET and DAO in serum in model group was increased, indicating the colonic mucosa seriously damaged and the intestinal permeability increase. Using Matrine and APS mixed with Matrine could reduce the colonic mucosa injury and the colonic mucosal permeability insignificantly (P<0.05or P<0.01).4. Expression of tight junction protein ZO-land Occludin in intestinal tissue of UC rats and the effect of the herb monomer.Observed the expression of ZO-1in intestinal tissue of UC rats with the method of western blot while ovserved the distribution and expression of Occludin in the colonic mucosa epithelium tight junction regions by immunohistochemistry.Results:Compared with normal group, the expression of tight junction protein ZO-land Occludin in colonic mucosa tissue of UC model group rats significantly decreased (P<0.05or P<0.01). At the end of2nd week, the expression of tight junction protein ZO-1and Occludin in all treatment group was increased, but there was no statistically significant difference between groups. At the end of4th week, compared with the model group, the expression of tight junction protein ZO-1and Occludin in Matrine and APS mixed group was significantly increased (P<0.05or P<0.01) and the expression of tight junction protein Occludin in APS group was also increased (P<0.05).5. Expression of tight junction protein ZO-land Occludin in lung tissue of UC rats and the effect of the herb monomer. Observed the expression of ZO-1in lung of UC rats with the method of western blot while ovserved the distribution and expression of Occludin in the rat alveolar epithelium tight junction regions by immunohistochemistry.Results:Compared with normal group, the expression of tight junction protein ZO-1and Occludin in lung tissue of UC model group rats significantly decreased (P<0.05or P<0.01) At the end of2nd,4th week, the expression of tight junction protein ZO-1in lung tissue in all treatment group was increased. At the end of2nd week, the expression of tight junction protein ZO-1in lung tissue in the two mixed group was significantly increased (P<0.05). At the end of4th week, the OD values of tight junction protein ZO-1in lung tissue in APS group was obviously higher than normal group and model group, but there was no statistically significant difference among all groups. The expression of tight junction protein Occludin in lung tissue in APS group and the two mixed group was increased significantly (P<0.05).6. The changes of inflammatory cytokine and oxygen free radical in lung of UC rats and the effect of the herb monomer.The content of TNF-α and IL-1β in lung was determined by ELISA while the activity of SOD, MDA and MPO was evaluated with colorimetry.Results:At the end of2nd,4th week, compared with normal group, the level of TNF-a, IL-1β, MDA, MPO in lung in UC rats was increased significantly while the activity of SOD was decreased significantly (P<0.05or P<0.01). The herb monomer could relieve lung injury of UC rats by promoting the capacity of the lung to resist inflammation and eliminate oxygen free radical, reducing lipid peroxidation.7Effect of the herb monomer on the expression of TFF3in lung and intestinal tissue of UC rats.Obvserved the distribution and expression of TFF3in lung and intestinal tissue by immunohistochemistry.Results:At the end of2nd,4th week, compared with normal group, the expression of TFF3in intestinal tissue was decreased significantly (P<0.05) while the expression of TFF3in intestinal tissue in the two mixed group was increased significantly (P<0.05). At the end of2nd,4th week, the expression of TFF3in lung tissue had no statistically significant difference among all groups (P>0.05).Conclusions1. Setting up the rat UC model by allergizing with colonic mucosal of rabbit added TNBS-50%alcohol coloclysis could produce the symptoms similar to human clinical manifestation of UC and reflect intestinal barrier dysfunction and immune imbalance the both two contributing factors of UC. It is an ideal model for studying the pathogenesis of UC.2. The pathomorphological observation of colonic mucosal and lung tissue of UC rats showed massive infiltration of inflammatory cells, hyperemia, dropsy, vasculitis and intestinal fibrosis in later stages, providing pathological evidence for the theory of "toxin impairing intestine collateral and then upward attack the lung"3. The expression of tight junction protein in colonic mucosal tissue was decreased, indicated the structure of colonic mucosa epithelium tight junction injury, causing the colonic mucosal permeability insignificantly increased, having colonic mucosal barrier dysfunction. It is maybe the foundation of "toxin impairing intestine collateral" pathogenesis theory.4. The expression of tight junction protein in lung tissue of UC rats was decreased and this was considered closely related to the lung injury of UC rats. It maybe the foundation of the theory of "toxin impairing intestine collateral and then upward attack the lung" and also maybe the key cause for the persistent unhealed and relapses of UC.5. The abnormal expression of inflammatory cytokine and oxygen free radical in lung of UC rats may be the common material basis between lung and large intestine in UC, also could be seen as the "endogenous toxin" causing UC and lung injury of UC.6. The herb monomer Matrine, APS and the two mixed can improve the general life station of UC rats, relieve the pathomorphological changes of colonic mucosal and lung effectively. APS has advantage in putting on weight of UC rats, while Matrine and Matrine mixed with APS can reduce the disease activity of UC rats.7. The herb monomer Matrine, APS and the two mixed is able to reduce inflammation in lung and colonic mucosal, improve local microcirculation and thus repair damaged lung and intestine tissue by increasing the expression of tight junction proteins, repairing damaged lung and intestine barrier, decreasing the lung and intestinal permeability, resisting to inflammation and oxygen free radical and upregulating the expression of repair factor in lung and intestine. |
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