苏黄止咳胶囊改善哮喘豚鼠气道重塑的作用及其机制

Effects and mechanism of Suhuang Antitussive Capsules in relieving airway remodeling of asthmatic guinea pigs

目的探讨苏黄止咳胶囊改善哮喘豚鼠气道重塑的作用及其机制。方法36只豚鼠随机分为正常组,模型组,苏黄止咳胶囊低、中、高剂量组(1.5、3、6 g/kg),阳性对照组(沙丁胺醇,5 mg/kg)。除正常组外,各组豚鼠均腹腔注射1 mg/mL卵清白蛋白(OVA)和10 mg/mL Al(OH)_3混悬液1 mL,同时以1%OVA雾化致敏,隔天刺激1次制造哮喘模型,连续给药2周。末次给药后1 h,检测豚鼠的咳嗽潜伏期,HE染色分析豚鼠气管及肺组织病理形态,ELISA测定肺组织中白介素IL-4、IL-5和组胺水平,immunoblot法检测肺组织中相关蛋白变化,离体实验观察苏黄止咳胶囊对豚鼠支气管舒张率的影响,MTT比色法检测苏黄止咳胶囊(125、250、500μg/mL)和阳性对照组(氨茶碱,50μg/mL)对组胺诱导大鼠支气管平滑肌细胞(RBSMCs)细胞增殖的抑制率影响,免疫荧光法检测其对增殖细胞核抗原(PCNA)蛋白的影响,immunoblot法检测其对酪氨酸激酶(JAK2)、信号传导与转录激活因子(STAT3)、蛋白激酶(Akt)、细胞周期相关蛋白的影响。结果与模型组比较,苏黄止咳胶囊延长哮喘豚鼠的咳喘潜伏期,改善气管平滑肌增厚、肺组织炎症及肺组织IL-4、IL-5、组胺水平;改善离体豚鼠支气管舒张率;有效抑制RBSMCs细胞增殖,改善PCNA、JAK2、STAT3、Akt及周期相关蛋白表达。结论苏黄止咳胶囊通过影响JAK/STAT信号通路来调控细胞周期相关蛋白抑制RBSMCs细胞增殖,改善支气管平滑肌增生,缓解气道重塑,从而发挥较好的平喘作用。

AIM To investigate the effects and mechanism of Suhuang Antitussive Capsules in relieving airway remodeling of asthmatic guinea pigs.METHODS Thirty-six guinea pigs were randomly divided into blank group,model group,Suhuang Antitussive Capsules groups(1.5 g/kg low-dose,3 g/kg medium-dose,6 g/kg high-dose),and salbutamol group(5 mg/kg).Except for the blank group,the other groups had their guinea pigs induced into asthma models given 2-week consecutive administration of intraperitoneal injection of a mixture solution of 1 mL 1 mg/mL OVA and 10 mg/mL Al(OH)3,and 1%OVA stimulation every other day.At 1 h after the last administration,the guinea pigs were subjected to the detection of their cough latency;the pathological observation of the lung and trachea tissues by HE staining;the measurement of the levels of inflammatory factors IL-4,IL-5 and histamine by ELISA;and determination of several related proteins expressions in the lung tissue by immunoblot;the verification of the effects of Suhuang Antitussive Capsules on bronchodiastolic rate in guinea pigs;the identification of in vitro effects of Suhuang Antitussive Capsules(125,250,500μg/mL)and aminophylline(50μg/mL)on proliferation activity in histamine-induced RBSMCs by MTT,the capsules’effect on PCNA expressions by immunofluorescence,and on the expression of JAK2,STAT3,Akt and cell cycle-related proteins by immunoblot.RESULTS Compared with the model group,groups intervened with Suhuang Antitussive Capsules shared significantly prolonged cough latency,effectively relieved thickening of airway smooth muscle in trachea tissues,improved inflammation of lung tissue,and reduced levels of inflammatory factors IL-4,IL-5 and histamine in lung tissues;significantly improved bronchodiastolic rate in isolated guinea pigs.In contrast to the model group,Suhuang Antitussive Capsules groups displayed significantly inhibited RBSMCs proliferation,ameliorated expressions of PCNA,JAK2,STAT3,Akt and cell cycle-related proteins.CONCLUSION Suhuang Antitussive Capsules alleviate the asthma in guinea pigs via inhibiting RBSMCs proliferation,improving bronchial smooth muscle hyperplasia and airway remodeling,and regulating cell cycle-related proteins through JAK/STAT signal pathway.