| Airway hyperreactivity (AHR) is encountered frequently and manifested in many type of urgent or chronic respiratory tract diseases, such as asthma, chronic obstructive pulmonary disease, viral upper respiratory tract infection and so on. Accumulating works show that airway inflammation is the main pathogenesis of AHR, the drugs for this symptom are mainly aiming at anti-inflammatory and symptomatic treatment. Many problems have not been resolved though the AHR symptom can be treated or alleviated by some drugs, such as the recurrent attacks of AHR, airway remodeling, the deterioration of pulmonary function and so on. Clerodendrum bungei is used extensively in traditional medicine to treat superficial infection, furunculosis, mammitis, arthrositis, eczema, cephalalgia, odontalgia and so on. It is reported recently that the extracts of clerodendrum bungei have many pharmacological actions, including antagonizing tardive hypersensitivity, the inhibitory action of cellular immunity, soothing arthritis, soothing nephritis, treating malaria, relieving pain and mesmerizing, shrinking uterus. Therefore, clerodendrum bungei is possibly effective to treat or relieve AHR diseases.Objective: To investigate the potential of clerodendrum bungei as a novel drug aiming at treating AHR diseases.Methods: I .The pharmacologic action survey on total extracts of clerodendrum bungei:1. Extracting all the active components in clerodendrum bungei, and then processing to the praeparatum for oral use;2. Establishing a novel AHR mouse model by the way of ozone exposure, the details of means are listed as follows: place the mice in plastic boxes which are 40cm long, 30cm wide and 25cm high, and then influx in mixed gas with 2.0ppm ozone and fresh air, 20 minutes every day is needed to expose the mice in the mixed gas in 5 days consecutively.3. Divide all the mice into 5 groups: normal control group, medicine control group, model group, medicine therapy group and glucocorticosteroid therapy group.4. The therapeutic efficacy of clerodendrum bungei extracts is assessed by the criterions as follows: measuring the airway reactivity of mice, counting the number of white blood cell in bronchoalveolar lavage fluid(BALF) and classification, counting the number of white blood cell in peripheral blood and classification, detecting the content of lactate dehydrogenase in the supernatant of BALF, observing the degree of pathological changes of lung after making the pathological section of lung; Evaluating initially the toxic action of clerodendrum bungei extracts through observing the pathological sections from nephridial tissue and hepatic tissue of mice and counting the number of white blood cell in peripheral blood.5. Observing the influence of blood serum which contain clerodendrum bungei extracts on the proliferation of human bronchial epithelial cells (HBECs) by the way of blood serum pharmacological. II. The pharmacologic action survey on three types of active constitutents in extracts of clerodendrum bungei:1. Extracting and separating three active constitutents (A component, B component and C component), and pretesting the components of them; and then processing them to three types of praeparatums for oral use respectively.2. Establishing a novel AHR mouse model by the way of ozone exposure(the means are the same as above),the three types of praeparatums were administered to the mouse model in three different groups by the way of intragastric;3. Divide all the mice into 5 groups: normal control group, model group, A component therapy group, B component therapy group and C component therapy group.4. the therapeutic efficacy of clerodendrum bungei extracts is assessed by the criterions as follows: measuring the airway reactivity of mice, counting the number of white blood cell in bronchoalveolar lavage fluid (BALF), counting the number of white blood cell in peripheral blood, detecting the content of total protein in the supernatant of BALF, observing the degree of pathological changes of lung after making the pathological section of lung.Results: I .The therapeutic efficacy of total clerodendrum bungei extracts:1. The RL ratios of mice in model group when they were provoked by four different concentrations of methacholine are higher than the counterparts of mice in normal control group obviously (P<0.05), which illustrates that the AHR mouse model is made successfully. The RL ratios of mice in medicine group treated with clerodendrum bungei extracts are lower than the counterparts of mice in model group when they were provoked by four different concentrations of methacholine (P<0.05),the therapeutic effect of clerodendrum bungei is almost equal to that of glucocorticosteroid.2. The number of white blood cell in BALF of mice in model group are more than that of mice in normal control group, and the number of white blood cell in BALF of mice in medicine group are less than that of mice in model group (P<0.05).It is found that the granulocyte and lymphocyte in BALF increase in number, while the administrating of clerodendrum bungei extracts can decrease the number of granulocyte and lymphocyte in BALF which were increased due to ozone exposure (P<0.05).3.The number of white blood cell in peripheral blood of mice in model group are more than that of mice in normal control group, and the number of white blood cell in peripheral blood of mice in medicine group are less than that of mice in model group(P<0.05). It is found that the type of white blood cell in peripheral blood which increase in number are neutrophilic granulocyte, lymphocyte, monocyte and eosinophile granulocyte, while the administrating of clerodendrum bungei extracts can decrease the number of neutrophilic granulocyte, lymphocyte, monocyte and eosinophile granulocyte in peripheral blood which were increased due to ozone exposure(P<0.05).4. When it comes to the content of lactic acid dehydrogenase(LDH) in BALF, ozone exposure can increase the content of LDH in BALF, while the treatment of clerodendrum bungei extracts can decrease it which would increase due to ozone exposure.5.It is shown from the HE dyeing of lung pathological section of mice in each group, the pathological changes of lung in model group are more severe than that of lung in normal control group(P<0.05),while the treatment of clerodendrum bungei extracts can mitigate the degree of pathological changes(P<0.05).(the indexes of pathological changes are judged by shedding of bronchial epithelial cells, thickening of bronchial tubal wall, thickening of smooth muscle, the content of endocritic in bronchus, the infiltration of inflammatory cell at the location of bronchus and vessel nearby and the pathological changes of pulmonary alveoli).6. All the pathological sections of nephridial tissue and hepatic tissue were stained by HE dyestuff, there were shown under microscope that all the structures of nephridial tissue and hepatic tissue were normal without any obvious inflammation or injury, which indicates that clerodendrum bungei extracts have not obvious toxic or secondary action to nephridial tissue and hepatic tissue. The difference of the number of white blood cell in peripheral blood between medicine control group and normal control group is insignificant (P>0.05), which illustrates that clerodendrum bungei extracts have no leukocyte-inhibition or obvious systemic toxic action.7. As can be seen from the MTT growth curves, the viable count in all the three groups which were processed with blood serum containing multiplication concentration of clerodendrum bungei extracts are more than the group which were processed with blood serum without any clerodendrum bungei extracts when 12 hours later, 36 hours later,48 hours later,3 days later,4 days later and 5 days later after the admoveaturing of different type of blood serum, Which hints that the blood serum with clerodendrum bungei extracts may have positive action to the proliferation of HBECs.II. The therapeutic efficacy of three types of active constitutents in clerodendrum bungei extracts:1. The RL ratios of mice in model group when they were provoked by four different concentrations of methacholine are higher than the counterparts of mice in normal control group obviously (P<0.05), which illustrates that the AHR mouse model is made successfully. The RL ratios of mice in C component therapy group and A component therapy group which were treated with C active constitutent and A active constitutent in clerodendrum bungei extracts are lower than the counterparts of mice in model group when they were provoked by four different concentrations of methacholine (P<0.05), While the RL ratios of mice in B component therapy group which were treated with B active constitutent in clerodendrum bungei extracts are lower than the counterparts of mice in model group when they were provoked by 0.78mg/ml concentrations of methacholine (P<0.05),but there are no statistical significance when they were provoked by other three concentrations of methacholine(0.39mg/ml,1.56mg/ml,3.12mg/ml).2. The number of white blood cell in BALF of mice in model group are more than that of mice in normal control group, and the number of white blood cell in BALF of mice in C component therapy group and A component therapy group are less than that of mice in model group (P<0.05). While there are no statistical significance when the number of white blood cell in BALF of mice in B component therapy group are compared with that of mice in model group.3. The number of white blood cell in peripheral blood of mice in model group are more than that of mice in normal control group, and the number of white blood cell in peripheral blood of mice in all the three groups which are treated with three different active constitutents are less than that of mice in model group (P<0.05).4. The total protein content in BALF can reflect the severity of lung injury. In this study, the total protein content in model group increase, while the total protein content in C component therapy group are less than that in model group obviously.5. It is shown from the HE dyeing of lung pathological section of mice in each group, the pathological changes of lung in model group are more severe than that of lung in normal control group (P<0.05), and the pathological changes of lung in C component therapy group and A component therapy group are more gentle than that of lung in model group (P<0.05), While there are no statistical significance when the pathological changes of lung in B component therapy group are compared with that of lung in model group (P>0.05).Conclusion: 1. Ozone exposure can cause the occurrence of airway inflammatory reaction, AHR and the pathological changes of airway and pulmonary alveoli on mice. Clerodendrum bungei extracts can mitigate the pathological changes of airway and pulmonary alveoli, inflammatory reaction and airway hyperreactivity on mice caused by ozone exposure.2. The blood serum with clerodendrum bungei extracts may have positive action to the proliferation of HBECs.3. C active constituent in clerodendrum bungei can mitigate airway inflammatory reaction and the pathological changes of airway and pulmonary alveoli due to ozone exposure, lower the degree of AHR significantly, is possibly an effective component to treat AHR diseases.
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