疏风通络方对哮喘小鼠模型Eotaxin、CCR3蛋白表达及ERK磷酸化的影响  

Effect of Shufeng Tongluo Prescription on Eotaxin and CCR3 Protein Expression and ERK Phosphorylation Level of Asthma Mice

作　　者：刘超武[1] 王洁 熊桅 王悦 朱振刚[1] LIU Chaowu;WANG Jie;XIONG Wei;WANG Yue;ZHU Zhengang(National Clinical Research Center of Chinese Acupuncture and Moxibustion,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;Tangshan Municipal Hospital of Traditional Chinese Medicine,Tangshan 063000,China)

机构地区：[1]天津中医药大学第一附属医院,国家中医针灸临床医学研究中心,天津300381 [2]唐山市中医院,河北唐山063000

出　　处：《中国实验方剂学杂志》2022年第14期54-60,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81673900)。

摘　　要：目的:初步探讨疏风通络方通过影响哮喘小鼠模型血清嗜酸细胞活化趋化因子(Eotaxin)及肺组织CC类趋化因子受体3(CCR3)、细胞外信号调节激酶(ERK)磷酸化的表达水平进而抑制哮喘气道炎症的机制。方法:将70只C57BL/6小鼠随机分成正常组、哮喘模型组、疏风通络方低、中、高剂量组(7.75、15.5、30 g·kg^(-1))、百日咳素(PTX)组、CCR3抑制剂(SB328437)组、磷脂酰肌醇3-激酶抑制剂(LY294002)组、p38蛋白激酶拮抗剂抑制剂(SB203580)组、ERK抑制剂(PD98059)组。采用卵清蛋白(OVA)、氢氧化铝[Al(OH)3]腹腔注射+OVA雾化建立小鼠哮喘模型(均为0.2 mL)。造模成功后,应用苏木素-伊红(HE)染色观察小鼠各组肺组织炎症浸润情况、酶联免疫吸附测定法(ELISA)检测各组小鼠血清中的Eotaxin[CC类趋化因子11(CCL11)、CC类趋化因子24(CCL24)]的水平;蛋白免疫印迹法(Western blot)检测肺组织ERK磷酸化水平及CCR3含量。结果:与正常组比较,模型小鼠支气管明显收缩,管腔狭窄,肺泡结构破坏,肺组织中可见大量炎性细胞浸润,支气管内可见黏液栓,气管黏膜下组织水肿,皱襞增多等表现,小鼠血清中CCL11、CCL24的含量显著增加(P<0.01),肺组织中CCR3蛋白的表达量明显增高(P<0.05),模型组、PTX组肺组织中的ERK水平明显升高(P<0.05),模型组、疏风通络方低剂量组肺组织中磷酸化(p)-ERK的水平明显升高(P<0.05);与模型组比较,病理显示疏风通络方高剂量组肺组织病变明显减轻;疏风通络方高剂量组、SB328437组CCL11含量明显降低(P<0.05),疏风通络方低、高剂量组、PTX组、SB203580组、PD98059组、SB328437组小鼠肺组织中CCR3蛋白的表达明显降低(P<0.05);疏风通络方高剂量组、PD98059组p-ERK的水平明显降低(P<0.05);PD98059组的ERK水平明显降低(P<0.05)。结论:疏风通络方可以抑制哮喘气道炎症,其机制可能与其通过下调CCR3蛋白、CCL11的表达及ERK磷酸化抑制嗜酸性�Objective: To preliminarily explore the mechanism of Shufeng Tongluo prescription(SFTLP)in inhibiting airway inflammation in asthma mice by affecting the expression levels of eotaxin in the serum, CC type chemokine receptor 3(CCR3), and extracellular signal-regulated kinase(ERK)phosphorylation in lung tissues. Method: Seventy C57BL/6 mice were randomly divided into a blank group,a model group,low-,medium-,and high-dose SFTLP groups(7.75,15.5,30 g·kg^(-1)),a pertussis toxin(PTX)group,a CCR3 inhibitor(SB328437)group,a phosphoinositide 3-kinase(PI3K)inhibitor(LY294002)group,a p38 protein kinase antagonist inhibitor(SB203580)group,and an ERK inhibitor(PD98059)group. The asthma model was induced in mice by intraperitoneal injection of ovalbumin(OVA)and aluminum hydroxide[Al(OH);]combined with OVA atomization(0.2 mL for all). After modeling,hematoxylin-eosin staining(HE staining) was used to observe the inflammatory infiltration of lung tissues in mice. Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of eotaxin[CC chemokine ligand(CCL)11and CCL24)in each group. Western blot was used to detect the levels of ERK phosphorylation and CCR3 in lung tissues. Result: Compared with the blank group,the model group showed obvious bronchial constriction,lumen stenosis,damaged alveolar structure,massive inflammatory cell infiltration in lung tissues,mucous plug in the bronchus,edema in the submucosal tissues of the trachea,increased folds,increased serum levels of CCL11 and CCL24(P<0.01),and increased expression of CCR3 protein in lung tissues(P<0.05). The ERK levels in lung tissues of the model group and the PTX group increased(P<0.05). The level of p-ERK in lung tissues of the model group and the low-dose SFTLP group increased(P<0.05). As revealed by pathological results,compared with the model group,the high-dose SFTLP group showed relieved lung lesions. The highdose SFTLP group and the SB328437 group showed reduced CCL11 content(P<0.05). The low-and high-dose SFTLP group,the PTX group,the SB203580 g

关 键 词：支气管哮喘 疏风通络方 CC类趋化因子11(CCL11) CC类趋化因子24(CCL24) CC类趋化因子受体3(CCR3) 细胞外信号调节激酶(ERK) 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R284R33