This study was aimed to exploreBi-Min-Gan (BMG) granules induced immune tolerance for allergic rhinitis cases through changes of Treg cells to achieve the goal of treatment. SD rats were used as experimental animal, which were divided into the blank group, model group and BMG granules group. Allergic rhinitis rat model was established. Intragastric administration of BMG granules was given. Levels of CD4+CD25+Foxp3+Treg and IL-4, IL-10, TGF-β1, IL-13 were detected with flow cytometry and ELISA assay. The results of flow cytometry showed that CD4+CD25+Foxp3+Treg decreased after the successful establishment of the model (P<0.05); but it increased after treatment of BMG granules (P<0.05) to reach the level of the normal blank group (P>0.05). ELISA showed the concentrations of IL-10 and TGF-β1 decreased significantly after the successful establishment of the model (P<0.0001); concentrations of IL-4 and IL-13 increased significantly (P<0.0001). While, after treatment of BMG granules, concentrations of IL-10 and TGF-β1 in blood rebound but did not reach the level of the normal blank group;concentrations of IL-4 and IL-13 in the blood decreased but did not reach the level of the normal blank group. It was concluded that BMG granules can upregulate the number of Treg cells in allergic rhinitis rat, increase the secretion of IL-10 and TGF-β1, inhibit Th2 cell activity, and reduce secretion of IL-4 and IL-13, so as to improve symptoms of allergic rhinitis.

OBJECTIVE: To investigate the mechanism. METHOD: We isolate the mouse bone marrow cells and cultured with rGM-CSF and rIL-4 to stimulate bone marrow cells to transfer to immature dendritic cells. And then the immature dendritic cells were costimulated with ILPS and different concentrations of Bimingan Granule. RESULT: MHC II, CD80, CD86 were detected by flow cytometer and TNF-alpha, IL-1beta, IL-6 and IL-10 were quantified by ELISA. CONCLUSION Our results showed that Bimingan Granule may significantly inhibit the differentiation of immature dendritic cells to mature dendritic cells.
Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Mice ; Mice, Inbred C57BL ; Rhinitis, Allergic ; drug therapy ; immunology

Objective We aimed to (i) observe the therapeutic effect of Yiqi Wenyang Prescription (YQWYP, which consists of milkvetch root, tangshen, dried ginger, cassig twig, ephedra, biond magnolia flower-bud, Chinese magnoliavine fruit, earthworm, and liquorice root) on an allergic rhinitis (AR) murine model and the protective effect on the tight junction (TJ) key proteins zonula occludens-1 (ZO-1) and occludin and (ii) elucidate whether YQWYP exerts these effects by inhibiting autophagy in nasal mucosal cells.Methods Thirty C57BL/6 mice were randomly divided into six groups with 5 mice per group: (i) the blank group, (ii) the model group, (iii-v) the YQWYP low-dose, mid-dose, and high-dose groups(7, 14, 28 g/kg), and (vi) the cetirizine group(1.667 mg/kg). The model was established by intraperitoneal injection and nasal stimulation of ovalbumin. 1 hour before the nasal stimulation, mice in the blank group or in the model group were given normal saline by gavage, mice in the YQWYP groups and those in the cetirizine group were given the corresponding medicines solution by gavage. After 7 days, mice behavior was scored. After that, the mice were sacrificed and the nasal mucosa tissues were harvested. The inflammatory reaction of the nasal mucosa was observed by HE staining, the proliferation of goblet cells in the nasal mucosa was observed by PAS staining, the IgE content in nasal lavage fluid was determined by ELISA, the expression levels of ZO-1 and occludin were analyzed by immunohistochemistry, and the expression levels of LC3B, P62, and Beclin1 in the nasal mucosa of the blank group, model group, and YQWYP high-dose group were analyzed by Western blotting.Results After the successful establishment of the AR model, compared with the blank group, the symptom scores of nose scratching, sneezing, and runny nose in the model group were higher, while the total scores was significantly higher(P<0.01). In the model group, the nasal mucosal epithelium was disrupted and destroyed, a large number of inflammatory cells were infiltrated, goblet cells showed hyperplasia, mucus production was increased, mucosal swelling was obvious, the amount of IgE in nasal lavage fluid was increased (P<0.01). The protein expression levels of ZO-1 and occludin were decreased (P<0.05), the protein expression levels of LC3BⅡ/Ⅰ and Beclin1 were increased (P<0.05), P62 protein expression was decreased (P<0.05). After medication intervention, compared with the model group, in the YQWYP high-dose group and the cetirizine group, the total score were reduced (P<0.01), the swelling of the nasal mucosa was reduced, the infiltration of inflammatory cells was inhibited, the number of goblet cells was decreased, the amount of IgE in nasal lavage fluid was reduced(P<0.01), the protein expression of ZO-1 and occludin were increased (P<0.05), the protein levels of LC3BII/I and Beclin1 were decreased (P<0.05), P62 protein expression was increased (P<0.05). Conclusion YQWYP has a good therapeutic effect on AR mice, and its mechanism may be related to the promotion of nasal mucosal repair by inhibiting autophagy of nasal mucosal cells.

OBJECTIVE：To preliminarily s tudy the potential mechanism of astragaloside Ⅳ on allergic rhinitis （AR）model mice. METHODS ：C57/BL6 mice were randomly divided into blank group ，model group and astragaloside Ⅳ group，with 10 mice in each group. Except for blank group ，AR model was prepared by sensitization and challenge with ovalbumin on day 0，7，14 and 21-27. Astragaloside Ⅳ group was given astragaloside Ⅳ 40 mg/kg intraperitoneally at the dose of 0.02 mL/g on the 15th to 27th day of modeling （given the drug 1 h before challenge sensitization on the 21st to 27th day ）. Blank group and model group were given constant volume of normal saline intraperitoneally ，once a day. Twenty-four hours after sensitization from the last challenge ， the infiltration of inflammatory cells in the nasal mucosa of each group was observed ，and the contents of interleukin 4（IL-4）， IL-5 and interferon gamma （IFN-γ）in the nasal lavage fluid were measured. The levels of reactive oxygen species （ROS），and the count of phosphorylated Janus kinase 2（p-JAK2）and phosphorylation signal transduction and activation of transcription protein 6 （p-STAT6）positive cells in the nasal mucosa and spleen as well as the phosphorylation levels of JAK 2 and STAT 6 proteins in spleen tissue （i.e. p-JAK 2/JAK2 ratio，p-STAT6/STAT6 ratio）were also determined. RESULTS ：Compared with blank group ，the number of inflammatory cells in the nasal mucosa （eosinophils and mast cells ）in the model group ，the contents of IL- 4 and IL- 5 in the nasal lavage fluid ，and the levels of ROS in the nasal mucosa and spleen tissues in the model group ，the count of p-JAK 2 and p-STAT 6 positive cells increased significantly ，the p-JAK2/JAK2 ratio，p-STAT6/STAT6 ratio in the spleen tissue were significantly increased （P＜0.05），and the content of INF-γ in the nasal lavage fluid was significantly decreased（P＜0.05）. Compared with model group ，the count of inflammatory cells infiltrated in the nasal mucosa ，the contents of IL- 4 and IL- 5 in the nasal cavity lavage fluid ，the level of ROS and the number of p-JAK 2 and p-STAT 6 positive cellsin the nasal mucosa and spleen tissue as well as the p-JAK2/JAK2 ratio and p-STAT 6/STAT6 ratio in spleen tissue were decreased significantly （P＜0.05），and the content of INF-γ in nasal lavage fluid was significantly increased（P＜0.05）. CO NCLUSIONS：Astragaloside Ⅳ can effectively improve the inflammatory response in AR model mice ，the mechanism of which may be related to down-regulation of JAK2/STAT6 signaling pathway and ROS level.

Objective: To observe the effect of Yiqi Wenyang Decoction on the infiltration and activation of NK cells in nasal mucosa of mouse model with allergic rhinitis (AR), and to explore the potential mechanism for effective intervention of AR with Yiqi Wenyang Decoction. Methods: Fourty-eight mice were randomly divided into blank group, model group, low, medium and high dose of Yiqi Wenyang Decoction group and Cetirizine group, with 8 rats in each group. After modeling of AR, the model group was filled with 0.9% sodium chloride solution. Yiqi Wenyang Decoction groups of each dose were given different concentrations of Yiqi Wenyang Decoction water extract, while the Cetirizine group was given aqueous solution of Cetirizine. The behavior, morphological changes of nasal mucosa and infiltration of NK cells in nasal mucosa were observed. The levels of IL-4 and INF-γ in nasal lavage fluid were measured. Besides, the drug safety was observed by acute toxicity test. Results: In the respect of behavioral scoring, middle and high dose of Yiqi Wenyang Decoction group were superior to the model group (number of sneezing: q value was 7.189, 8.748, respectively; number of scratching nose: q value was 12.074, 14.560, respectively; all P<0.05). In middle and high dose of Yiqi Wenyang Decoction group, the infiltration of NK cells and nasal lavage fluid IL-4 levels were lower than those in model group (IOD: q value was 10.073, 12.322, respectively; IOD/Area: q value was 10.954, 14.073, respectively; IL-4: q value was 4.705, 6.801, respectively; all P<0.05). There was no significant difference in nasal lavage fluid of INF-γ among each group (Fv=1.166, P>0.05). In acute toxicity test, no obvious poisoning symptoms and death occurred in mice. Conclusion Yiqi Wenyang Decoction can control the nasal symptom, reduce the local NK cell infiltration of nasal mucosa and inhibit the expression of the 2-type cytokines released by NK cells, which may be related with the potential mechanism of effective intervention of AR with Yiqi Wenyang Decoction.