[Effects of fermented white mustard seed plaster on airway immune balance and its inflammatory response mechanism in bronchial asthma rats]

Abstract

Objectives: To observe the effects of fermented white mustard seed plaster on the balance of T helper cell (Th) 1/Th2, Th17/T regulatory cell (Treg) immune balance in bronchial asthma (BA) rats, so as to explore the possible mechanism of fermented white mustard seed plaster in the treatment of BA.

Methods: Healthy SD rats were randomly divided into blank, model, medication and acupoint application groups, with 10 rats in each group. BA model was prepared using ovalbumin sensitization and atomization. Rats in the acupoint application group were treated with fermented white mustard seed plaster at "Dazhui" (GV14), bilateral "Feishu" (BL13) and "Fengmen" (BL12) for 4 h;rats in the medication group were given intraperitoneal injection of 1 mg/kg dexamethasone sodium phosphate;both groups were treated once daily for 14 consecutive days. Behavioral observations and scoring were conducted on rats in each group before and after intervention. HE staining was performed to observe the pathological morphological changes of lung tissue. ELISA was used to detect the contents of interferon γ (IFN-γ), interleukin (IL)-4, IL-17, IL-10, tumor necrosis factor α (TNF-α), and immunoglobulin E (IgE) in serum. real-time quantitive PCR was used to detect the mRNA levels of T-box transcription factor (T-bet), GATA binding protein 3 (GATA-3), retinoic acid-related orphan receptor γt (RORγt), and forkhead box protein 3 (Foxp3) in lung tissue. Western blot was used to detect the protein expression levels of T-bet, GATA-3, RORγt, and Foxp3 in lung tissue.

Results: After treatment, compared with the blank group, the behavioral score was significantly increased (P<0.01);the thickness of bronchial wall muscle layer increased with a large amount of inflammatory cell infiltration, thickening of alveolar wall, abnormal lung tissue structure, and severe lung consolidation;the contents of serum IL-4, IL-17, TNF-α, and IgE were significantly increased (P<0.01), while the contents of serum IFN-γ and IL-10 were significantly decreased (P<0.01);the mRNA and protein expression levels of GATA-3 and RORγt in lung tissue were increased (P<0.01), while the mRNA and protein expression levels of T-bet and Foxp3 in lung tissue were decreased (P<0.01) of rats in the model group. Compared with the model group, the behavioral score was significantly decreased (P<0.01);the pathological morphological damage of lung tissue was significantly improved;the contents of serum IL-4, IL-17, TNF-α, and IgE were significantly decreased (P<0.01, P<0.05), while the contents of serum IFN-γ and IL-10 were significantly increased (P<0.01, P<0.05);the mRNA and protein expression levels of T-bet and Foxp3 in lung tissue were increased (P<0.01), while the mRNA and protein expression levels of GATA-3 and RORγt in lung tissue were decreased (P<0.01) of rats in the medication and acupoint application groups. Compared with the medication group, the contents of serum IL-4 were significantly increased (P<0.05);the mRNA and protein expression levels of GATA-3 and RORγt in lung tissue were significantly increased (P<0.01, P<0.05), while the mRNA and protein expression levels of T-bet and Foxp3 were significantly decreased (P<0.01, P<0.05) of rats in the acupoint application group.

Conclusions: Fermented white mustard seed plaster can alleviate airway inflammation in BA, and its mechanism may be achieved by up-regulating the expression of T-bet and Foxp3, down-regulating the expression of GATA-3 and RORγt, promoting Th1/Th2, Th17/Treg immune balance, and reducing the secretion of related pro-inflammatory factors to exert a therapeutic effect on asthma.

Keywords: Airway inflammation; Bronchial asthma; Fermented white mustard seed plaster; Th1/Th2, Th17/Treg.