| Objective: Through the establishment of bronchial asthma guinea pig model, to observethe effect of Chaipu decoction on eosinophil apoptosis and regulation of expression of Fas,Fas-L, Bcl-2in the lung tissues of asthmatic guinea pig,and to explore the possiblemechanism of Chaipu decoction promoting eosinophil apoptosis for bronchial asthma.Methods:40Healthy male guinea pigs were randomly divided into four groups, Normalcontrol group, Asthma model group, Dexamethasone group, Chaipu decoction group,(N=10), repeatedly sensitizing and stimulating guinea pig model of asthma, by Ovalbumin(OVA). After the last inhalation recorded behavioral signs score for each group of guineapigs symptoms, tested each group of guinea pigs blood eosinophil number and bronchialalveolar lavage fluid white blood cell count, eosinophil number, percentage of eosinophilcell, Hematoxylin-eosin staining was observed in each group of guinea pigbroncho-pulmonary pathological change; tested eosinophil apoptosis of each group guineapig lung tissues in the TUNEL method; each group Fas, Fas-L, and Bcl-2proteinexpression were measured by Immunohistochemistry, Western blot, and each group thelevel of Fas, Fas-L, Bcl-2mRNA expression were measured by situ hybridization andRT-PCR.Results:1. Bronchial asthma symptom score of each group: Compared with the asthma model group,asthma symptoms of dexamethasone group and Chaipu decoction group significantly wereimproved and the asthma symptom scores decreased significantly (P <0.05).2. Peripheral blood eosinophil cell count of each group: Compared with normal controlgroup, the eosinophil count was significantly higher in the peripheral blood of asthmatic model group, the difference was significant (P <0.01); compared with the asthma modelgroup and dexamethasone group, peripheral blood eosinophil counts of Chaipu decoctionwere significantly lower (P <0.05); there is no significant difference betweendexamethasone group and Chaipu decoction group,(P>0.05).3. BALF leukocyte count, the proportion of eosinophils, eosinophil count of each group:White blood cell count, eosinophil proportion, eosinophil cell count in BALF of the asthmamodel group were higher than normal control group, the difference was statisticallysignificant (P <0.01); dexamethasone group, Chaipu decoction group two groups, BALF ofwhite blood cell count, the proportion of eosinophils, eosinophil cell counts were lowerthan the asthma model group (P <0.05), but higher than the normal control group (P <0.05).4. Lung tissues pathological changes of each group: Normal bronchial lung tissue wasnormal integrity and smooth lumen airway mucosal edema, epithelial cells no obvioushyperplasia, occasional eosinophils, a complete normal alveolar septa. Asthma model groupbronchial stenosis deformation, visible mucosal edema, epithelial cell hyperplasia,shedding, mucus secretion was significantly increased proliferation of bronchial smoothmuscle hypertrophy, infiltration of the bronchial mucous layer and perivascularinflammatory cells consisting mainly of eosinophils, alveolar septal thickening, structuraldisorder. Bronchopulmonary pathological changes of dexamethasone group and Chaipudecoction group were significantly lighter than the asthma model group.5. EOS apoptosis of lung tissues in each group: Compared with the normal control group,there are a large number of eosinophilic granulocytes cell infiltration in lung tissues ofasthmatic model group, eosinophil apoptosis index was significantly lower (P <0.01);compared with the asthma model group: dexamethasone group and Chaipu decoction groupapoptotic eosinophils of dexamethasone group and Chaipu decoction group significantlyincreased,and the apoptotic index was significantly higher (P <0.01); there was nostatistically significant difference between the dexamethasone group and the Chaipudecoction group (P>0.05).6. Expression of Fas, Fas-L, Bcl-2protein in each group: Immunohistochemical resultsshowed that compared with the asthma model group, expression of Fas, Fas-L protein of the other two groups were increased significantly (P <0.05); compared with normal group:Bcl-2protein expression of the asthma model group was significantly increased (P <0.01)in the dexamethasone group and Chaipu decoction group; Western blot results showed thatcompared with the asthma model group, Fas, Fas-L protein expression was significantlyincreased in dexamethasone group, and Chaipu decoction group (P <0.01); comparedwith the normal control group, the Bcl-2protein expression was significantly increased inasthmatic model group (P <0.01);and compared with model group the data of the othertwo groups was significantly lower (P <0.05). Compared with there was no significantstatistical difference between dexamethasone group and Chaipu decoction group (P>0.05).7. Expression of Fas, Fas-, Bcl-2mRNA in Guinea pig lung and bronchus tissues: In situhybridization results showed that: Compared with the asthma model group,bothdexamethasone group and Chaipu decoction group the expression of Fas mRNA, Fas-LmRNA were significantly increased (P <0.05); Bcl-2mRNA expression of asthma modelgroup was significantly increased (P <0.01); compared with the other two groups， themodel group was significantly lower (P <0.05). RT-PCR results showed that: the expressionof Fas mRNA, Fas-L mRNA expression in the dexamethasone group and Chaipu decoctiongroup were increased significantly (P <0.01), compared with the normal control group, thestatistical difference was also increased significantly (P <0.05); compared with thenormal control group, the Bcl-2mRNA expression of the asthma model group wassignificantly increased (P <0.01); compared with the model group, the data of other twogroups was significantly lower (P <0.05). There was no significant statistical differencebetween dexamethasone group and Chaipu decoction group (P>0.05).Conclusion:1. Chaipu decoction could significantly reduce eosinophil cell count in asthma guinea pigperipheral blood and bronchoalveolar lavage fluid, alleviate the inflammation of airwayand the lung tissues of asthmatic gunea pigs.2. Chaipu decoction could promote apoptosis of eosinophils in the lung tissues of asthmaticguinea pigs.3. Chaipu decoction could contribute to the expression of Fas and Fas-L in asthma guinea pig lung tissues, and reduce Bcl-2expression level. |
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