| ObjectiveObserving the effect on the airway inflammation and airway remodeling and the changes of the expression c LTD4, IL-17 after the intervention of Montelukast and Huang Qi Huai in the rat model of chronic asthma; To explore the mechanism of Montelukast combined with Huang Qi Huai in the treatment of asthma.Methods1. Forty male Sprague-D wley (SD) rats aged 4-6 weeks and weighted 120±10g were choosen and divided randomly into five groups: control group(Control),model group(Model),Montelukast group(MK),Huang Qi Huai group(H),Montelukast+Huang Qi Huai group(MK+H), esch group has 8 rats.2. On the 1st day and the 8th day, the other four groups except the control group were sensitized to be injected intraperitioneally with 0.1%OVA (including 10% aluminium hydroxide and the inactive pertussis bacillus)1ml; On the 15th day, they were challenged with 1% OVA (8ml) through Nebulized Pump, twenty minutes for each time, once every second day, lasting four weeks to built the chronic rat asthma model. the control group were sensitized and challenged by the physiological saline. The other treatment groups were admmistered intragastrically with Montelukast (1mg/100g/d). Huang Qi Huai (0.4g/100g/d) and Montelukast (1mg/100g/d)+Huang Qi Huai(0.4g/100g/d) respectively.3. All the animals were sacrificed and collected samples within 24 hours after the last chanllenging. Get plasma,bronchoalveolar lavage fluid (BALF) and superior lobe of right lung tissue. Cuperior lobe of right lung tissue,routinely dehydrated,embedded in paraff, measure the expression of IL-17 in lung tissue by immunohistochemistry, The airway inflammation and remodeling were analyzed by histochemistry staining with HE, Masson and AB-PAS.Total cells score and differential score in BALF were counted.The levels of LTD4 ane IL-17 in the plasma and BALF were measured by Enzyme-linked immunosorbent assay(ELISA).Results1.General observation:compared with group Control, Rat of group Model appeare gradually irritability, scratching the nose, shortness of breath, breathing nodding cyanosis, cyanosis, eating little or quiescence, piloerection even depilation,Delayed reaction. group Control had no such symptoms. The symptoms of Group MK, group H, group MK+H was reduced in different degree compared with group Model, group MK and group MK+H improved significantly.2. Histopathological:1)The degree of inflammatory cell around the airway:compared with the Control group, the degree of inflammatory cell around the airway in the model group were significantly higher(P<0.05), compared with group Model, Group MK, group H and group MK+H were significantly ameliorated (P<0.05), group MK compared with Group H were ameliorated (P<0.05),group MK+H compared with Group MK were significantly ameliorated (P<0.05).2)The percentage of goblet cells accounting for the epithelia cell:group MK, group H and group MK+H were significantly lower than group Model respectively (P< 0.05), group MK+H and group MK were significantly lower than group H(P<0.05), group MK compared with Group MK+H,the difference had no statistical significance.3) The deposition of collagen:compared with group Model, group MK and group MK+H were significantly ameliorated (P<0.05), group MK+H compared with Group MK, group H compared with group Model,the difference had no statistical significance.4) The area of smooth muscle:group MK, group H and group MK+H were significantly lower than group Model respectively (P<0.05), Group MK+H was significantly lower than group MK and group H(P<0.05), group MK compared with group H,the difference had no statistical significance.5) Immunohistochemistry:the expression of IL-17 in lung tissue of group MK,group H and group MK+H were significantly lower than group Model respectively (P<0.05), group MK+H was significantly lower than group MK and group H(P<0.05).3. The number of total cells,differential cell count:The number of total cells, the percentages of EOS and the percentages of NEU were increased compared to group Control (P<0.05); group H, group MK and group MK+H were decreased step by step compared to group Model,but group H compared with group Model the difference had no statistical significance about the percentages of NEU, group MK+H compared with group MK,the difference had no statistical significance about the percentages of EOS and the percentages of NEU.4. The concentration of LTD4, IL-17 in plasma and BALF:1) The concentration of L.TD4 in plasma and BALF:group Model were increased compared to group Control (P<0.05);in the plasma,group MK and group MK+H were decreased compared to group Model (P<0.05), group H compared with group Model and group MK+H compared with group MK,the difference had no statistical significance;in BALF, group MK,group H and group MK+H were decreased compared to group Model (P<0.05), group MK+H were decreased compared to group H(P<0.05), group MK+H compared with group MK,the difference had no statistical significance.2) The concentration of IL-17 in plasma and BALF:group Model were increased compared to group Control (P<0.05), group MK and group MK+H were decreased compared to group Model (P<0.05), group MK+H were decreased compared to group MK(P<0.05),group H compared with group Model,the difference had no statistical significance.5. Correlation analysis showed that the level of IL-17 in the plasma and BALF was positively correlated with the level of IL-17 in the lung tissue (P<0.05). The percentages of EOS and NEU was positively correlated with the expression of IL-17 in lung tissue (P<0.05)Conclusion1. The contents of LTD4 in blood plasma and BALF were increased in the asthmatic rat.2. The contents of IL-17 in blood plasma and BALF were increa,ed in the asthmatic rat. IL-17 was significantly highly expressed in the small bronchial columnar epithelium cells and inflammatory cells by immunohistochemical method.3. Correlation analysis showed that the level of IL-17 in the plasma was positively correlated with the level of IL-17 in the BALF and the lung tissue. Suggesting the test of IL-17 in the plasma is a simple and accurate methed for detecting the expression of IL-17 in lung tissue.4. Montelukast can reduce the concentration of LTD4, IL-17 in the plasma and BALF, reduce the expression of IL-17 in lung tissue, improve the airway inflammation and the airway remodeling in the asthmatic rat. Combined Huang Qi Huai adjuvant montelukast treatment of asthmatic rat, the parameters mentioned above were further improved, down regulation the expression of LTD4 and IL-17 was probably one of the mechanisms about anti-inflammatory.,... |
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