In Vivo Study On The Mechanism Of The Ethanol Extract Of Root Of Anacyclus Pyrethrum On Cough Variant Asthma

Objective:To study the anti-inflammatory and anti-oxidative mechanism of ethanol extract of root of Anacyclus Pyrethrum?EEAP?on cough variant asthma?CVA?model rats.Methods:Ninety rats were randomly divided into control group?drinking water?,model group,high-dose EEAP group(640 mg·kg^-1),medium-dose group(320 mg·kg^-1),low-dose group(160 mg·kg^-1)and prednisone acetate group(2.5 mg·kg^-1).Rat model of cough variant asthma was induced by ovalbumin?OVA?.Comparing cough times of rats,observing pathological changes of lung tissue,counting inflammatory cells in alveolar lavage fluid by Swiss staining method.The serum levels of interleukin-4?IL-4?,interleukin-5?IL-5?,interleukin-6?IL-6?,interleukin-13?IL-13?,reactive oxygen species?ROS?,granulocyte macrophage colony stimulating factor?GM-CSF?,tumor necrosis factor?TNF-??,interferon-??IFN-??and superoxide dismutase?SOD?and malondialdehyde?MDA?in rats were measured by ELISA,and dendritic cells were detected by flow cytometry.The expression of costimulatory molecules CD80 and CD86on the surface of cells?DCs?and the expression of TLR4 and NF-?B?p65?in lung and kidney homogenate of rats in each group were detected by real-time fluorescence quantitative PCR?RT-PCR?.Western Blot method was used to detect the expression of TLR4,NF-?B?p 65?,Trx,GSK-3?,?-GSK-3???-catenin,?-?-catenin and 4-HNE in lung and kidney homogenate of rats in each group.Results:Compared with the normal group,EEAP could reduce the cough frequency and the infiltration of inflammatory cells in the lungs of CVA rats.At the same time,the levels of SOD and IFN-?were significantly increased in each dose group of EEAP?P<0.05?,but the levels of IL-4,IL-5?IL-6?IL-13?ROS?GM-CSF?TNF-?and MDA were not significantly different from those of prednisone acetate group?P<0.05?;In addition,the expressions of CD 80 and CD 86costimulatory molecules on DC surface were significantly decreased in each dose group of EEAP?P<0.05?;EEAP could significantly reduce the expression of TLR4 and NF-?B?p65?in TLR4/NF-?B?p65?signaling pathway?P<0.05?,and significantly reduce the protein expression levels of?-GSK-3???-catenin and 4-HNE in Wnt signaling pathway?P<0.05?and Trx,GSK-3?and?-?-catenin were significantly increased?P<0.05?.There was no significant difference among the groups?P>0.05?.Conclusion:EEAP may play an anti-inflammatory and anti-asthmatic role by inhibiting the expression of IL-4?IL-5?IL-6?IL-13?ROS?GM-CSF?TNF-??MDA,TNF-?and increasing the levels of IFN-?and SOD,down-regulating the co-stimulatory molecules CD80 and CD86 on DC surface,It may play an anti-inflammatory and anti-asthmatic role by and down-regulating the expression of TLR4/NF-?B?p65?signal pathway TLR4 and NF-?B?p65?and Wnt signaling pathway?-GSK-3?,?-catenin and 4-HNE and up-regulating Trx,GSK-3? and ?-?-catenin.