基于Wnt/β-catenin信号通路探讨白茅根多糖对哮喘气道重塑的作用  

The improving effect of Rhizoma imperatae polysaccharide on airway remodeling in asthma via Wnt/β-catenin signaling pathway

作　　者：杜宁[1] 刘爱华 冯艳 王莹莹 张娜 DU Ning;LIU Ai-hua;FENG Yan;WANG Ying-ying;ZHANG Na(Department of Pediatrics,Baoding City Maternal and Child Health Hospital,Baoding 071000,Hebei,China)

机构地区：[1]保定市妇幼保健院儿科,河北保定071000 [2]泊头市医院儿科,河北沧州062150 [3]邢台市临城县中医医院儿科,河北邢台054300 [4]河北省盐山县人民医院儿科,河北沧州061300

出　　处：《广东医学》2023年第4期441-447,共7页Guangdong Medical Journal

基　　金：河北省中医药管理局科研项目(2021277)。

摘　　要：目的基于Wnt/β-连环蛋白(β-catenin)信号通路探讨白茅根多糖对哮喘气道重塑的改善作用及机制.方法选无特定病原体(specific pathogen free,SPF)级雄性Sprague Dawley(SD)大鼠35只,随机分为5组:空白组(生理盐水处理)、模型组、地塞米松组、白茅根多糖低剂量组、白茅根多糖高剂量组,每组7只.卵清蛋白致敏诱导哮喘大鼠模型,造模成功后,分为模型组(生理盐水)、地塞米松组(0.5 mg/kg)、白茅根多糖低剂量组(10 mg/kg)、白茅根多糖高剂量组(20 mg/kg),灌胃给药治疗,连续8周.采用苏木素-伊红(HE)染色观察肺组织病理变化;免疫组组织化学法检测肺组织α-平滑肌肌动蛋白(α-SMA)的表达;瑞士-吉姆萨染色分类计数支气管肺泡灌洗液中的炎症细胞数;酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-4、IL-5、IL-13含量及血清中免疫球蛋白E(IgE)、TNF-α、IL-6含量;吸入乙酰甲胆碱测定大鼠气道高反应性;实时荧光定量聚合酶链式反应(qRT-PCR)/蛋白免疫印迹(Western blot)检测肺组织Wnt5a、Wnt7a、β-catenin、糖原合成酶激酶3β(GSK-3β)mRNA或蛋白表达.结果与空白组相比,模型组肺泡隔增厚伴炎细胞浸润,炎性细胞浸润评分、中性粒细胞、淋巴细胞数、气道阻力及TNF-α、IL-4、IL-5、IL-13、IgE、IL-6含量明显升高,肺组织α-SMA及Wnt5a、Wnt7a、β-catenin、GSK-3βmRNA和蛋白表达明显升高(P<0.05);与模型组相比,白茅根多糖高剂量组能显著抑制哮喘中性粒细胞、淋巴细胞数,降低气道阻力及TNF-α、IL-4、IL-5、IL-13、IgE、IL-6含量,并抑制肺组织α-SMA及Wnt5a、Wnt7a、β-catenin、GSK-3βmRNA和蛋白表达(P<0.05).结论白茅根多糖可能通过下调Wnt/β-catenin信号通路的激活来改善哮喘大鼠气道重塑,具有抑制哮喘的效果.Objective To investigate the ameliorative effect and mechanism of Rhizoma imperatae polysaccharide via Wnt/Beta-catenin(β-catenin)signaling pathway on asthma airway remodeling.Methods An ovalbumin sensitization-induced asthma rat model was treated by gavage in the dexamethasone group(0.5 mg/kg),and the low-dose group(10 mg/kg)and the high-dose group(20 mg/kg)of Rhizoma imperatae polysaccharide for 8 weeks.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of lung tissue.Immunohistochemistry was used to detect the expression of alpha-smooth muscle actin(α-SMA)in lung tissue.Swiss-Gimza staining was used to sort and count the number of inflammatory cells in bronchoalveolar lavage fluid.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),interleukin-5(IL-5)and interleukin-13(IL-13)in bronchoalveolar lavage fluid and the contents of immunoglobulin E(IgE),TNF-αand interleukin-6(IL-6)in serum.Inhaled acetylcholine was used to determine airway hyperresponsiveness in mice.Quantitative real-time fluorescence polymerase chain reaction(qRT-PCR)/Western blot were applied to assess the Wnt5a,Wnt7a,β-catenin,Glycogen synthase kinase 3β(GSK-3β)mRNA or protein expression in lung tissues.Results Compared with the control group,alveolar septal thickening with inflammatory cell infiltration,inflammatory cell infiltration score,neutrophil,lymphocyte count,airway resistance and TNF-α,IL-4,IL-5,IL-13,IgE,IL-6 contents were significantly higher,and lung tissueα-SMA and Wnt5a,Wnt7a,β-catenin,GSK-3βmRNA and protein expression were significantly elevated in the model group(P<0.05).Compared with the model group,the high-dose Rhizoma imperatae polysaccharide significantly inhibited asthmatic neutrophil and lymphocyte counts;reduced airway resistance and TNF-α,IL-4,IL-5,IL-13,IgE,IL-6 content;and suppressed lung tissueα-SMA and Wnt5a,Wnt7a,β-catenin,GSK-3βmRNA and protein expression(P<0.05).Conclusion Rhizoma imperatae polysaccha

关 键 词：哮喘 白茅根多糖 气道重塑 WNT/Β-CATENIN信号通路 

分 类 号：R563.8[医药卫生—呼吸系统] R285.5[医药卫生—内科学]