| Objective: Promoting blood circulation and dispersing phlegm method that has often used in clinical, and Catalpol is one of the effective components in the water extraction of HTHXF. Our previous study found that HTHXF and Catalpol have a certain therapeutic effect on OVA-induced asthma mice, and the mechanism may be through intervention of the Notch signal transduction, inhibiting the abnormal activation of Th2 cells, regulates Th1/Th2 imbalance, increase the secretion of interferon(IN-γ) and reduce the interleukin-4 and interleukin(IL-4), interleukin-17A(IL-17A) in BALF of asthmactic mice. This subject is to observe the effect of HTHXF and Catalpol on inflamlmation and mucins MUC5 AC in asthmatica mice in OVA-induced asthma mice, and explore its mechanism.Methods: 1. Asthmatic model of mice was induced by intraperitoneal injection of Ovalbumin(OVA), and then atomization inhalation of 2% OVA aroused in Balb/c mice. Mice were sensitized by an intraperitoneal injection of OVA on the1 st,8th and 15 thdays, and subsequent Ly were stimulated to the nebulization with 2% OVA from the 21 th day to the 27 th day. Mice of the treatment group were given HTHXF and Catalpol through the mouth from the 9th day to the 27 th day. The allergic inflammatory response and the expression of MUC5 AC were observed on the 28 th day. Record the latency and the symptoms of the asthmatic mice from the 25 th day to the 27 th day during the period of aerosolization with OVA; 2. Airway resistance(RI) was measured 24 h later after the last time of stimulation by the Buxco’s modular and invasive system; 3. Histo Logica L inflammatory assessment and the hyperplasia changes of the goblet cell in lung tissues were evaluated by the hematoxylin and eosin(HE), mucus production was measured by periodic acid-Schiff staining(PAS); 4. Record the transformation of the counts of the inflammatory cells of the Bronchoalveolar Lavage(BALF) through Wright,s-Giemsa staining; 5. Interleukin-5(IL-5), MUC5 AC, interleukin-10(IL-10) and TGF-β1(TGF-β1) in BALF were evaluated by enzyme-linked immunosorbent assay(ELISA); 6. The viability of the effect of the Catalpol on 16 HBE cells induced by EGF was evaluated by MTT assay; 7. The effect of Catalpol on the expression of MUC5 AC in the supernatant of cell culture was evaluated by ELISA;Results: 1. All groups of mice in good condition before aerosolization, but in the process of OVA challenged process, there are appeared some symptoms of asthma, such as in continence, irritability, hair rising, shortness of breath. With aerosolized sometimes later, the incubation period of asthma in normal mice significantly longer than the model group mice(P < 0.01).Meanwhile, the asthmatic mice lose their weight, and the hair lost their luster. While compared with model group,the treatment groups of HTHXF and Catalpol can markedly relieve the symptoms of asthma and decrease latency of asthmatic mice(P < 0.01); 2. Every group of RI have increased with the increasing dose of Meth(0、 6.25、 12.5、25、50 mg/m L). Meanwhile, the RI value of model group has obviously higher than that of the control group(P<0.05 or P<0.01). However, Dexamethasone and high dose of Catalpol treatment group could obviously decreased AHR compared with model group in response to Meth, especially at the dose of 25 mg/m L,50mg/m L(P<0.05 or P<0.01). The value of RI has reduced in different degree after given different dose of HTHXF and Catalpol(P<0.05 or P<0.01); 3. A remarkable infiltration of inflammatory cells into peribronchiolar, perivascular connective tissues; thickening of basement membrane, airway mucosa epithelial shedding, mucosal epithelium incomplete, hyperplasia of airway epithelial cells and mucus hypersecretion were observed in asthmatic mice compared with control group. According to the pathologic results of graded and positive area, total score of model group around the trachea, peribronchiolar, perivascular have significantly higher than the normal group, and the positive area of model group has significantly greater than the normal group(P < 0.01). The pathologic results of graded and positive area, total score have reduced in different degree after given different dose of HTHXF and Catalpol when compared with model group; 4. Model group challenged with OVA, white total cells, neutrophils and macrophages in BALF have significantly increased, and they have considered to be statistically significant compared with control group(P<0.01). OVA challenged mice treated with HTHXF and Catalpol showed markedly reduced leukocytosis, a fall in absolute eosinophilia and lymphocytosis when compared with model group(P<0.05 or P<0.01); 5. Compared with control group, content of IL-5 and MUC5 AC in BALF of asthmatic mice have obviously higher in model group, and the content of IL-5 and MUC5 AC have obviously reduced after given different dose of HTHXF and Catalpol when compared with model group. Compared with control group, the levels of TGF-β1 and IL-10 in BALF of asthmatic mice have obviously lower than model group, and the levels of TGF-β1 and IL-10 have obviously elevated after given different dose of HTHXF and Catalpol when compared with model group(P<0.05 or P<0.01); 6. Compared with model group, Catalpol(0.3μM、1μM、3μM、10μM、30μM、100μM、300μM) have no effect on the viability of 16 HBE cell; 7. Compared with control group, content of MUC5 AC in 16 HBE cells culture supernatant was obviously higher in model group, and the content of MUC5 AC has obviously reduced after given different dose of Catalpol(3μM、10μM、30μM) when compared with model group(P<0.01).Conclusion: 1. We successly established the model of asthma through estimate the incubation period, HE staining and PAS staining; 2. HTHXF and Catalpol can obviously reduce the contents of leukocytosis,and inflammatory cells such as eosinophilia, lymphocytosis and neutrophils in BALF of asthmatic mice; 3. HTHXF and Catalpol can reduce the value of RI in asthmatic mice; 4. HTHXF and Catalpol can relieve inflammatory cells infiltration, thickening of basement membrane,mucus hypersecretion and goblet cell hyperplasia in lung tissues of asthmatic mice; 5. HTHXF and Catalpol can reduce the levels of IL-5,MUC5 AC and increase the levels of TGF-β1 and IL-10 in BALF of asthmatic mice; 6. HTHXF and Catalpol have certain inhibitory effect on MUC5 AC, and its mechanism may be through the regulation of inflammation cells and regulatory factor; 7. Catalpol(0.3μM、1μM、3μM、10μM、30μM、100μM、300μM) have no effect on the viability 16 HBE cells; 8. Catalpol(3μM、10μM、30μM) have certain inhibitory effect on MUC5 AC in cell culture supernatant of EGF induced 16 HBE cells. |
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