The Effect Of Thrombin On Airway Inflammation Of The Rat With Asthma

Objective:The bronchial asthma is a respiratory disease, which is common in children. When the bronchial asthma broke out, the patient would have the symptom of wheeze, anhelation, chest tightness, cough, and so on. Because of the asthma, the patient suffered from great pain and heavy economic berden. IL-6is an inflammation factor involved in asthma, which plays an important role in asthma airway inflammation. In recent years, some studies showed thrombin can promote the activity of the pro-inflammatory NF-κB, then thrombin can mediate the airway inflammation. This experiment have prepared asthma model. Through①Detect the activity of the thrombin in the BALF of the rat using the method ELISA;②Observe the infiltration of inflammatory cell in the lung by hematoxylin-eosin staining;③Test the expression of IL-6in the lung by immunohistochemisty;④Test the expression of NF-κB in the lung by the method of western blot. Then we discuss the influence of thrombin on airway inflammation of the rat with asthma and our conclusions may provide new path for discussing the mechanism of asthma.Methods:24adult female rats were randomly divided into four groups:normal control group(N group), asthma group(A group), thrombin group(T group), and thrombin inhibitors group(H group), there were6rats in each group. We make the asthmatic model by sensitizing and stimulating the rats with ovalbumin. The rats were injected ovalbumin in the inner of each thigh and whooping cough bacili levitation liquid in the intraperitoneal as an adjuvant on the1、8days. From the15day, the rats were recevied inhalation of2%OVA for30minites one time, three times a week for8weeks. Before ultrasonic atomizing inhalation, each rat in the T group was given72u thrombin through inhalation. While in the H group each rat was given hirudin for500u/kg through inhalation. In the normal group, we used saline water instead of ovalbumin during the intervention. When finished, we take the lung and BALF from the rat for the following experiment:①Detect the activity of the thrombin in the BALF of the rat using the method ELISA;②Observe the infiltration of inflammatory cell in the lung by hematoxylin-eosin staining;③Test the expression of IL-6in the lung by immunohistochemisty;④Test the expression of NF-κB in the lung by the method of western blot.Results:1. Behavior change of the experimental ratsThe experimental groups have the appearance of hair mats. In the process of making model, the rats appear cough varying degresses, sneeze, tachypnea. abdominal muscle rigid, quadriplegia, costive, slow in reacting, and so on. While the control group were normal.2.The activity of the thrombinThe activity of the thrombin in the asthma group is higher than that in the control group. The activity of the thrombin in the thrombin group is higher than that in the asthma group. The activity of the thrombin in the thrombin inhibitors group is lower than that in the thrombin group.3. Infiltration of inflammatory cell in the lungHE staining showed that:There were no infiltration of inflammatory cell in the normal group, while the asthma group has infiltration of inflammtory cell. There were obvious infiltration of the inflammatory cells in the thrombin group, while the thrombin inhibitors group was alleviative than the thrombin group.4. Expression of IL-6in the lungThe expression of IL-6in lung were measured by immunhistochemistry. There was hardly any espression of IL-6in the normal group. The espression of IL-6were increased in the asthma group. IL-6was obviously expressed in the lung tissue of thrombin group. And the espression of IL-6in the thrombin inhibitors group was fewer than the thrombin group.5. NF-κB expression in the lungWe detect the expression of the NF-κB using the method of western blot. There was higher expression in the asthma group than that in the normal group. The expression of NF-κB in thrombin group reached the peak in the four groups, while the level of NF-κB in thrombin inhibitors group was decreased, and lower than the thrombin group.Conclusions:1.The asthma model was successfully established.2.The level of the thrombin in the airway of the rats with asthma rised. Thrombin can promote the infiltration of inflammatory cell in the lung.3.Thrombin can up-regulate the expression of NF-κB in airway of the rats with asthma, promote the expression of IL-6, thus participate in asthma inflammation.