Protective effect of Xiaochuangning Granules on Th1/Th2 balance and STAT1 phosphorylation in asthma rats
SHAN Minmin1 LI Changan2 CUI Hongsheng3
1.Beijing University of Traditional Chinese Medicine, Beijing 100029, China;
2.Department of Pulmonary Disease,Dongzhimen Hospital of Beijing University of Traditional Chinese Medicine, Beijing 100029, China;
3.Department of Pneumology, the Third Affiliated Hospital of Beijing University of Traditional Chinese Medicine, Beijing 100029, China
Abstract：Objective To observe the effect of Xiaochuangning Granules on Th1/Th2 balance and STAT1 pathway in asthma, and to explore its anti-Xiaochuangning mechanism. Methods Fourty SPF male SD rats in total, using the digital random method randomly divided into 5 groups, the normal group, asthma group, IL-27 prevention, Xiaochuangning Granules group and the asthma rather pellet + Fludarabine (STAT1 inhibitors) group, each group of eight rats, were established by intraperitoneal injection of ovalbumin joint aluminum hydroxide sensitization and intranasal ovalbumin excitation method were established. Each group was treated with IL-27 nasal drops (IL-27 prevention group) in the pre-sensitization stage, or by gavage of Xiaochuangning Granules (Xiaochuangning Granules group) in the excitation stage, or combined with intraperitoneal injection of the STAT1 inhibitor Fludarabine to observe the effect. Rats in each group were sacrificed after the last stimulation, serum and BALF were collected, the expression levels of IL-4 and IFN-γ were detected by ELISA, and the expression levels of pSTAT1 (STAT1 phosphorylated protein), signal transduction and transcriptional activator 1 (STAT1) in lung tissues were detected by Western blot. Results Compared with the model rats in asthma group, the IL-4 contents in serum and BALF were significantly decreased and the IFN-γ contents were significantly increased in Xiaochuangning Granules group and IL-27 prevention group (P ＜ 0.05). Compared with Xiaochuangning Granules group, Fludarabine inhibited the therapeutic effect of Xiaochuangning Granules (P ＜ 0.05). Compared with the asthma group model rats, the expression level of pSTAT1 protein in the lung tissues of the Xiaochuangning Granules group and the IL-27 prevention group was significantly increased (P ＜ 0.05), and Fludarabine inhibited the expression of pSTAT1 protein in the lung tissues of the Xiaochuangning Granules group (P ＜ 0.05). Conclusion Xiaochuangning Granules can effectively improve Th1/Th2 imbalance in Xiaochuangning rats, and up-regulate the level of inhibited pSTAT1 protein and increase the phosphorylation of the STAT1 pathway. The mechanism of asthma treatment may be related to the IL-27 induced phosphorylation of the STAT1 pathway.
[1] 沈华浩，杜旭菲，应颂敏.新版中国支气管哮喘防治指南与全球支气管哮喘防治创议的异同[J].中华结核和呼吸杂志，2018，41（3）：166-168.
[2] 苏楠，林江涛，刘国梁，等.我国8省市支气管哮喘患者控制水平的流行病学调查[J].中华内科杂志，2014，53（8）：601-606.
[3] 崔红生，武维屏，赵燕荣，等.哮喘宁煎剂治疗支气管哮喘急性发作期的临床研究[J].北京中医药大学学报，1999（1）：57-60.
[4] Hu C，Li Z，Feng J，et al. Glucocorticoids Modulate Th1 and Th2 Responses in Xiaochuangning Mouse Models by Inhibition of Notch1 Signaling [J]. Int Arch Allergy Immunol，2018，175（1/2）：44-52.
[5] 任传云，武维屏，周冠芬，等.哮喘宁颗粒剂对哮喘大鼠糖皮质激素受体的影响[J].中国中医药信息杂志，2004， 11（2）：118-119.
[6] 中华医学会呼吸病学分会哮喘学组.支气管哮喘防治指南（2016年版）[J].中华结核和呼吸杂志，2016，39（9）：675-697.
[7] 谢华健，周小敏，林奇栋，等.哮喘患者血清Th1/Th2类细胞因子水平变化及意义[J].山东医药，2011，51（31）：59-60.
[8] Loureno O，Mafalda Fonseca A，Taborda-Barata L. T cells in sputum of Xiaochuangning patients are activated independently of disease severity or control [J]. Allergol Immunopathol，2009，37（6）：285-292.
[9] Nakanishi K，Tsutsui H，Yoshimoto T. Importance of IL/-18-induced super Th1 cells for the development of allergic inflammation [J]. Allergol Int，2010，59（2）：137-141.
[10] 张红，顾猛.过敏性哮喘患儿血浆IFN-γ与IL-4及粪便双歧杆菌检测及分析[J].国际检验医学杂志，2015， 36（3）：367-368.
[11] Tsuchiya K，Isogai S，Tamaoka M，et al. Depletion of CD8+ T cells enhances airway remodelling in a rodent model of asthma [J]. Immunology，2009，126（1）：45-54. doi：10.1111/j.1365-2567.2008.02876.x.
[12] Matsukura S，Odaka M，Kurokawa M，et al. Transforming growth factor-β stimulates the expression of eotaxin/CC chemokine ligand 11 and its promoter activity through binding site for nuclear factor-κβ in airway smooth muscle cells [J]. Clin Exp Allergy，2010，40（5）：763-771. doi：10.1111/j.1365-2222.2010.03474.x.
[13] 何燕超，揭志军.白细胞介素27对哮喘免疫调控及治疗的研究进展[J].医学综述，2015，21（23）：4230-4233.
[14] Pflanz S，Timans JC，Cheung J，et al. IL-27，a heterodimeric cytokine composed of EBI3 and p28 protein，induces proliferation of naive CD4+ T cells [J]. Immunity，2002，16（6）：779-790.
[15] Artis D，Villarino A，Silverman M，et al. The IL-27 receptor （WSX-1） is an inhibitor of innate and adaptive elements of type 2 immunity [J]. J Immunol，2004，173（9）：5626-5634.
[16] Su X，Pan J，Bai F，et al. IL-27 attenuates airway inflammation in a mouse asthma model via the STAT1 and GADD45γ/p38 MAPK pathways [J]. J Transl Med，2016， 29，14（1）：283.
[17] 万晓朋，熊伟，张梅，等.IL-27对哮喘小鼠血清IgE的影响[J].成都医学院学报，2013，8（1）：23-25.
[18] Cao Y，Doodes PD，Glant TT，et al. IL-27 induces a Th1 immune response and susceptibility to experimental arthritis [J]. J Immunol，2008，180（2）：922-930.
[19] Villegas-Mendez A，de Souza JB，Lavelle SW，et al. IL-27 receptor signalling restricts the formation of pathogenic，terminally differentiated Th1 cells during malaria infection by repressing IL-12 dependent signals [J]. PLoS Pathog，2013，9（4）：e1 003 293. doi：10.1371/journal.ppat. 1003293.
[20] Lucas S，Ghilardi N，Li J，et al. IL-27 regulates IL-12 responsiveness of naive CD4+ T cells through Stat1-dependent and -independent mechanisms [J]. Proc Natl Acad Sci U S A，2003，100（25）：15047-15052.
[21] 赵云峰，梁栋，张梅，等.IL-27对哮喘小鼠气道炎症的影响[J].中国医药生物技术，2010，10（5）：357-360.
[22] Chen Z，Wang S，Erekosima N，et al. IL-4 confers resistance to IL-27-mediated suppression on CD4+ T cells by impairing STAT1 signaling [J]. J Allergy & Clinical Immunol，2013，132（4）：912-921.
[23] 黄伟强，李惠，袁梅，等.支气管哮喘患者外周血T细胞亚群Th1、Th2及其相关细胞因子水平变化及意义[J].山东医药，2015（20）：40-41.
2019, Vol. 16 
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