Supplementary MaterialsAdditional file 1. pathology, EOS matters in blood and bronchoalveolar lavage fluid were observed. The degree of the EOS apoptosis in rats was recognized. The manifestation Punicalagin ic50 content of interleukin (IL)-5, IL-10, chemokine (CCC motif) ligand 5 (CCL5), granulocyteCmacrophage colony-stimulating element (GM-CSF), transforming growth element beta 1 (TGF-1), interferon (IFN)-, and additional cytokines in rat serum and the genes of Eotaxin mRNA, Fas mRNA, FasL mRNA, Fas/FasL and Bcl-2 mRNA in the lung cells were determined. Results WTD can reduced airway resistance in rat models and improved airway compliance. The pathological changes of lung cells in WTD group were significantly alleviated, at the same time, WTD could reduce the EOS count in the blood and BALF smears of the asthmatic model rats. Compared with the model group, the apoptosis degree of EOS significantly improved in rats in the WTD group. The manifestation of IL-5, CCL5, and GM-CSF in the serum and the manifestation of Eotaxin mRNA, Bcl-2 mRNA in the lung cells in rats in the WTD group rats decreased. Moreover, the manifestation of IL-10, TGF-1, and IFN- in the serum and the expression of Fas mRNA, FasL mRNA in the lung tissues in rats in the WTD group rats increased compared with that in rats in the model group. Conclusions Wentong decoction may accelerate EOS apoptosis, reduce asthma inflammation, and alleviate the disease through regulating and controlling the factors related to the anti-apoptosis and pro-apoptosis. Electronic supplementary material The online version of this article (10.1186/s13020-018-0180-2) contains supplementary material, which is available to authorized users. for 15?min (20?C). After centrifuging, we recycled the eosinophilic cell layer of Percoll liquid interfaces with different densities. D-Hanks liquid was used for centrifugal washing at 1500?rpm for 10?min twice. Then, using D-Hanks liquid of 1 1?mL heavy suspension, we calculated the absolute numbers of EOS under a microscope. We adjusted the cell count to approximately 2??106/mL and took 1?mL for flow detection. Enzyme-linked immunosorbent assay (ELISA) We took 2?mL of the rats blood at room temperature, centrifuged at 3000?r/min for 15?min at static pressure for 2?h, and then stored at ??80?C after collecting the supernatant liquid. We performed the procedure in strict accordance with the ELISA Kit manual. We used the double antibody sandwich-ELISA assay test, to determine the IL-5, IL-10, GM-CSF, CCL5, TGF-, and IFN- (Abcam, Cambridge, UK) in the serum. RNA extraction and real-time polymerase chain reaction (PCR) We evaluated the effect of Wentong decoction on the expression of Eotaxin mRNA and Fas mRNA in the lung tissues of asthmatic rats by using real-time fluorescent quantitative PCR. We determined the primer sequences using glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and performed primer design (Table?1). We used Trizol total RNA extraction kit (DP405-02, Tiangen Biochemical Technology, Beijing Co., Ltd., Beijing, China) to extract total RNA from the 25?mg lung tissue according to the manufacturers instructions for inverse transcription of RNA samples to obtain the corresponding cDNA. After pretreatment of the upper machine mixture, we operated the real-time PCR device at 95?C, 30?s, and 40 PCR loops (collected the fluorescence at 95?C for 5?s and 60?C for 40?s). The target and internal control genes of each Punicalagin ic50 sample underwent real-time PCR, and the data were analyzed using 2?CT method. Punicalagin ic50 Tabel 1 antisense and Feeling primer sequences of Eotaxin, Bcl-2, Fas, FasL and GAPDH thead th align=”remaining” rowspan=”1″ colspan=”1″ Tools name /th th align=”remaining” rowspan=”1″ colspan=”1″ Primer series (5C3) /th th align=”remaining” rowspan=”1″ colspan=”1″ Item size (bp) /th /thead GAPDH upstream primerCCTTCCGTGTTCCTACCCC131GAPDH downstream primerGCCCAGGATGCCCTTTAGTGEotaxin upstream primerGCTACAAAAGAATCACCAACAACAG95Eotaxin downstream primerCTTTTTCTTGGGGTCAGCACAGBcl-2 upstream primerGGGCTACGAGTGGGATACTGGAG101Bcl-2 downstream primerCGGGCGTTCGGTTGCTCTFas upstream primerATCAATAATCATGGCTGTGT116Fas downstream primerTATTTGAGTGTATCCCTGCTFasL upstream primerGGTGCTGGTGGCTCTGGTT142FasL downstream primerTGTGCTGGGGTTGGCTATTT Open up in another window Data evaluation All data had been examined using SPSS software program (edition 17.0, SPSS, Inc., Chicago, IL, USA). The full total consequence of each group is shown in mean??regular deviation. em P /em ? ?0.05 indicates significant difference statistically. Solitary factor variance NewmanCKeuls and analysis test were useful for group analysis. If the info do not comply with regular distribution, the non-parametric KruskalCWallis check was useful ISGF-3 for assessment. Outcomes Lung function Lung function exam plays a significant role in analyzing the asthma intensity, prognosis, and curative aftereffect of drugs. To evaluate the effect of Wentong decoction in improving lung function in asthmatic rats, we tested the inspiratory resistance, expiratory resistance, and dynamic compliance of the airway in all groups of rats (Fig.?1). The results showed that compared with the normal group, the inspiratory resistance and expiratory resistance of rats in the model group increased significantly. Airway compliance significantly reduced, and the difference was statistically significant ( em p? /em ?0.05). After drug intervention, dexamethasone (DXM) and WTD significantly reduced inspiratory resistance and expiratory resistance.