Wuhu Decoction Influence For Respiratory Syncytial Virus Induced Young Asthmatic Rats NaiveCD4~+T Cell Differentiation

Objective:In this paper, the use of molecular biology, microscopic morphology, immunology and other research techniques and methods to study Wuhu decoction influence for respiratory syncytial virus induced young asthmatic rats NaiveCD4+T cell differentiation, in order to further clarify the Wuhu decoction anti-virus mechanism induced childhood asthma provide experimental evidence.Methods:60SD rat were randomly divided into blank group10and model group50. Making model adopts the method of OVA subcutaneous injection and intranasal RSV. After mice model established in model rats were randomly by blind method divided into bronchial asthma group dexamethasone group, Wuhu decoction high dose group, Wuhu decoction middle dose group and low dose group which each group with10. The first seven days in asthmatic medication group were given the appropriate liquid, blank control group and asthma model group were given the same volume of sterile distilled water,2times a day for14days. After the last administration, bronchoalveolar lavage fluid, serum, lung tissues and spleen tissues from all the rats were sacrificed. ELISA was used to detect serum IFN-y, IL-12, IL-10and IL-17concentrations; RT-PCR technique to detect expression for the spleen cell suspension, lung tissue transcription factor T-bet、GATA-3、RORγt、Foxp3mRNA; Western blot detection of lung tissue transcription factor T-bet, GATA-3, RORyt, Foxp3protein.Results:(1) Compared with the control group, the percent of model group total leukocytes, neutrophils, eosinophils was increased significantly.the difference was statistically significant (P<0.01); Compared with the asthma model group, Wuhu decoction in high dose group and middle dose group, the total number of white blood cells, neutrophils, eosinophils were decreased significantly, the difference was statistically significant (P<0.01), In lymphocytes has no significant changes.(2) Compared with the blank group, serum IFN-γ、IL-12concentration decreased, while IL-10、IL-17concentration (P<0.05) increased in model group; Compared with the model group, the Wuhu decoction high-dose group, serum IFN-γ、IL-12concentrations were significantly reduced, and IL-10、IL-17concentrations were significantly increased (P<0.01); middle dose group serum IFN-γ\IL-12concentration decreased, while IL-10、IL-17concentration (P<0.05) increased; low-dose group serum IFN-γ、IL-12、 IL-10、IL-17concentration change is not obvious.(3) Compared with the blank group, spleen cell suspensions T-bet、Foxp3mRNA expression decreased and GATA-3、RORyt mRNA expression increased (P<0.05) in model group; Compared with the model group, high dose group GATA-3、RORyt mRNA expression was significantly lower while T-bet、Foxp3mRNA expression was significantly increased (P<0.01); middle dose group GATA-3、RORyt mRNA was decreased and T-bet、Foxp3mRNA expression was increased (P<0.05); the low-dose group spleen cell suspension T-bet、GATA-3、RORyt and Foxp3mRNA expression has no change significantly.(4) Compared with the blank group, model group lung tissue T-bet、Foxp3mRNA expression decreased and GATA-3、RORyt mRNA expression increased (P<0.05); Compared with the model group, Wuhu decoction high dose group GATA-3、RORyt mRNA expression was significantly lower and T-bet、Foxp3mRNA expression was significantly increased (P<0.01); The middle dose group GATA-3、RORyt mRNA decreased while T-bet、Foxp3mRNA expression increased (P<0.05); The low-dose group T-bet、GATA-3、 RORyt and Foxp3mRNA expression has no change significantly.(5) Compared with the blank group, model group lung tissue T-bet、Foxp3protein expression decreased and GATA-3、RORyt protein increased (P<0.05); Compared with the model group, Wuhu decoction high dose group GATA-3、RORyt protein expression decreased significantly while T-bet、 Foxp3protein expression increased (P<0.01); The middle dose group GATA-3、RORyt protein decreased while T-bet, Foxp3protein expression increased (P<0.05).Conclusions:(1) The rats model which established by OVA intraperitoneal injection sensitization combine with OVA continuous aerosol inhalation and intranasal administration stimulate can be discovered asthma symptoms. And along with the Ach concentration increased the Re values increased.in bronchoalveolar lavage fluid, the total white blood cell is mainly eosinophils and neutrophils, airway mucosal inflammatory infiltration, which the above performance characteristics comply with asthma pathophysiology.(2) Wuhu decoction can up-regulation IFN-γ、IL-12and down-regulated IL-10、IL-17expression to regulate Th1/Th2、Th17/Treg imbalance cells, thus improving the state of immune disorders in asthmatic rats.(3) Wuhu decoction can up-regulation T-bet、Foxp3and also down-regulated GATA-3、RORyt expression to regulate Th1/Th2、Th17/Treg imbalance cells, control the reactions of Th2、Th17-type, thereby restoring immune asthmatic rats tolerated.