| Objective:To establish pulmonary disease (chronic bronchitis) and bowel disease (ulcerative colitis) animal models respectively, which are both chronic non-specific inflammatory disease. Select three different points, synchronous observe the respiratory and intestinal micro flora, MEK/ERK signal pathway, and its upstream mediators of inflammation receptor TNF-alpha receptor, IL-1beta receptors and downstream effecter substances C-FOS both in lung and large intestine, to explore the biological basis of the mass change of the lung and large intestine in pathological state.Method:Use smoked method to copy pulmonary disease (chronic bronchitis) rat model, take samples of lung and intestinal synchronous dynamic observation respectively at model20th,50th and70th. Use TNBS combined ethanol method to copy bowel disease (ulcerative colitis) rat model, take samples of lung and intestinal synchronous dynamic observation respectively at model8th,29th and50th. Detect the total number of aerobic bacteria, total number of anaerobic bacteria, E. coli, enterococci, staphylococcus aureus, clostridium perfringens, bifidobacterium and lactobacillus both in respiratory and intestinal at the same time. Detect MEK, p-MEK, ERK1/2, p-ERK1/2protein expression in the homogenate of lung tissue and colon tissue by Western-Blot method. Use ELISA method to detect the content of TNF-alpha receptor, IL-1beta receptor and C-FOS in the homogenates of lung tissue and colon tissue.Results:1. Respiratory and intestinal flora:Pulmonary disease (CB) model rats have different degree changes of flora species and quantities at each time point in the respiratory and intestinal. At model20th, the total number of aerobic bacteria in the respiratory and intestinal increased simultaneously. Intestinal microbial colonization resistance (B/E) decreased significantly. The total number of aerobic bacteria and staphylococcus aureus of respiratory and intestinal are moderate correlated (0.5<r<0.8). At model50th, the total number of anaerobic bacteria in respiratory and intestinal are moderate correlated. At model70th, the total number of aerobic bacteria and enterococci in respiratory and intestinal are moderate correlated.Bowel disease (UC) model rats appear intestinal flora imbalance, the number of probiotics decreased but opportunistic pathogen increased. At the same time, part of the respiratory tract micro flora is also have related change in certain degree. At model8th, the total number of aerobic and staphylococcus aureus in the respiratory and intestinal increased simultaneously, the total number of anaerobic and enterobacteriaceae increased in the intestine but decreased in the respiratory tract. At model29th, the total number of aerobic and staphylococcus aureus in the respiratory and intestinal reduced simultaneously, the total number of anaerobic and enterobacteriaceae reduced in the intestinal but increase in the respiratory tract. At model50th, the total number of aerobic bacteria, the total number of anaerobic bacteria and staphylococcus aureus in the respiratory and intestinal increased simultaneously.2. MEK/ERK signaling pathway:In pulmonary disease (CB) model rats, the contents of p-ERK1/2were significantly higher than the blank group in the lung tissues at model20th,50th and70th(P<0.05or P<0.01).The changes of MEK/p-MEK and ERK/p-ERK in the lung tissue is significant than the colon tissue. The change trend of MEK/p-MEK and ERK/p-ERK protein in the lung and colon tissue is more consistent.In bowel disease (UC) model rats, the content of ERK1/2and p-ERK1/2were significantly higher than the blank group in the colon tissue at model8th(P<0.01). The content of p-MEK was significantly higher than the blank group in colon tissue at model29th (P＜0.05). The ERK1/2and p-ERK1/2at model29th were significantly higher than the8th (P<0.05or P<0.01). The content of p-MEK, ERK1/2and p-ERKl/2were significantly reduced at model50th(P<0.05). Only in the model29th, the content of p-MEK in the lung tissue was significantly higher than the8th (P<0.05). The changes of MEK/ERK in colon tissue and lung tissue showed synchronization variation, or synchronous increased or reduced simultaneously, or increase in lung but decrease in intestinal, or decrease in lung but increase in intestinal.3. The content of TNF-alpha receptor, IL-1beta receptor and C-FOS:In pulmonary disease (CB) model rats, the content of TNF-alpha receptor and IL-1beta receptor in lung tissue were significantly reduced at model70th, but significantly increased in colon tissue at model70th (P<0.05or P<0.01). The content of C-Fos was significantly reduced both in lung and colon tissue at model50th (P<0.05).In bowel disease (UC) model rats, the content of TNF-alpha receptor was decreased in colon tissue but increased in lung tissue significantly at model8th (P＜0.05). At model8th and29th, the content of IL-1beta receptor was increased in colon tissue but decreased in lung tissue significantly (P＜0.05). The content of C-Fos was increased in colon tissue but decreased in lung tissue significantly at model29th and50th (P＜0.01).Conclusion:1. Pulmonary disease (CB) model rats’intestinal flora was changed. There showed disorders of micro-ecological system in respiratory and intestinal system when lung disease spread to the colon. There is a certain degree of dynamic correlation of respiratory and intestinal micro-ecological system. The micro-ecological of the lung and large intestine interaction may be one of the connotations of "the lung and the large intestine being interior-exteriorly related"2. Bowel disease (UC) model rats’respiratory tract flora was changed. When the bowel disease spread to the lung, part of respiratory and intestinal flora synchronization regular changed in bowel disease model rats, or synchronous increased, or reduced simultaneously. This showed that the microcosm’s flora changes may be one of the mechanisms and manifestations of "bowel disease spread to the lung" 3. Pulmonary disease rats have certain regular changes in the MEK/ERK pathway in the lung tissues. P-ERK1/2may be indicators of chronic bronchitis obvious changes in the MEK/ERK pathway. This prompts that MEK/ERK pathway may be one of the biological basis of "lung disease spread to the colon".4. Bowel disease rats showed similar change trends of MEK/p-MEK and ERK/p-ERK in the lung and colon tissue. To some extent reflects the synchronization between the changes in the lung and intestine. This prompts that MEK/ERK pathway may be one of the biological basis of "bowel disease spread to the lung".5. pulmonary disease rats’TNF-alpha receptor, IL-1beta receptor and c-Fos content appears synchronous dynamic change in the lung and intestine tissue, or decreased both in the lung and intestinal simultaneously, or decreased in the lung while increased in the intestine. This prompts that the contents of TNF-alpha receptor, IL-lbeta receptor and c-Fos may be one of the material bases of "lung disease spread to the colon".6. Bowel disease rats’TNF-alpha receptor, IL-1beta receptor and c-Fos were synchronous dynamic changed in the lung and intestine, or decreased both in the lung and intestinal simultaneously, or decreased in intestinal but increased in the lung, or increased in the intestine but decreased in the lung. This prompts that the contents of TNF-alpha receptor, IL-lbeta receptor and c-Fos may be one of the material bases of "bowel disease spread to the lung". |
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