爱罗咳喘宁对COPD大鼠p38MAPK,TNF-α及AQP5基因表达的影响

Effect of Ailuo Kechuanning on Expression of p38MAPK, TNF-α and AQP5 Genes in Rats with Chronic Obstructive Pulmonary Disease

摘要:

目的: 观察爱罗咳喘宁对慢性阻塞性肺疾病(chronic obstructive pulmonarydisease,COPD)大鼠模型支气管和肺组织中p38丝裂原活化蛋白激酶(p38mitogen activatein kinase,p38MAPK)、肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、及水通道蛋白5(aquaporin 5,AQP5)基因表达的影响,探讨爱罗咳喘宁方对COPD炎症及气道高分泌的作用及机制. 方法: 采用脂多糖(lipopolysaccharide,LPS)加烟雾诱导COPD大鼠模型,随机将大鼠分为正常组、模型组、爱罗咳喘宁低、中、高剂量组.正常组、模型组灌胃生理盐水(15.52 mL·kg^-1·d^-1),爱罗咳喘宁口服液低、中、高剂量组分别灌胃(7.75,15.52,31.04 g·kg^-1·d^-1),连续15 d.原位杂交、免疫组化检测p38MAPK、TNF-α及AQP5基因在细支气管和肺组织中的原位表达. 结果: 与正常对照组比较,模型组大鼠AQP5 mRNA[(4.52±0.11)VS(0.87±0.32),P<0.01]和蛋白[(3.98±0.22)VS(0.58±0.02),P<0.01]表达均显著减弱,而p38MAPK mRNA[(0.49±0.02)VS(0.85±0.02),P<0.01],TNF-α mRNA[(0.26±0.01)VS(0.44±0.01),P<0.01]均显著增强,p-p38MAPK蛋白[(0.30±0.01)VS(0.41±0.01),P<0.05]和TNF-α蛋白[(0.20±0.01VS(0.38±0.01),P<0.05]表达均显著增强,AQP5蛋白与p-p38MAPK,TNF-α蛋白表达均呈负相关(r分别为-0.547,-0.532,P<0.01);与模型组相比,爱罗咳喘宁中剂量组AQP5 mRNA[(0.87±0.32 VS 1.80±0.32,P<0.01)],AQP5蛋白[(0.58±0.02 VS 1.61±0.72,P<0.01)]表达显著增强,p38MAPK mRNA[(0.85±0.02)VS(0.51±0.02),P<0.01],TNF-α mRNA[(0.44±0.01)VS(0.40±0.01),P<0.01]表达减弱,p38MAPK蛋白[(0.41±0.01)VS(0.32±0.00),P<0.01]TNF-α蛋白[(0.38±0.01)VS(0.31±0.00), P<0.01]表达显著减弱.AQP5蛋白表达与p-P38MAPK,TNF-α蛋白均呈显著负相关(r分别为-0.542,-0.541,均P<0.01). 结论: 爱罗咳喘宁有抗炎和抑制COPD气道黏液高分泌作用,其机制可能与下调p38MAPK,TNF-α基因表达、上调AQP5基因表达有关.

Abstract:

Objective: To study the effect of Ailuo Kechuanning on expression of p38MAPK,tumor necrosis factor alpha(TNF-α),and aquaporin 5(AQP5) genes in rats with chronic obstructive pulmonary disease(COPD). Method: Lipopolysaccharide(LPS)smoke was used to COPD rat model,the experimental animals were randomly divided into normal control group,model group,Normal group, model group was given physiological saline(15.52 mL· kg^-1· d^-1) byintragastric, Ailuo Kechuanning group(7.75,15.52,31.04 g· kg^-1· d^-1),ig for 15 days. The expression of p38MAPK,TNF-α and AQP5 genes in the trachea, bronchus and lung tissue were detected by in situ hybridization and immunohistochemical techniques. Result: Compared with the normal group, the expression of mRNA and protein of AQP5 were weakened, but the expression of p38MAPK,TNF-α mRNA and protein were enhanced in model group, the expression of AQP5 protein was negatively correlated with p38MAPK(r=-0.547,P<0.01),TNF-α protein(r=-0.532,P<0.01).Compared Ailuo Kechuanning mediate-dose group with the model group, the expression of AQP5 gene in enhancements, However, the expression of p38MAPK,TNF-α genes was decreased apparently,the expression of AQP5 protein was negatively correlated with p38MAPK(r=-0.542,P<0.01),TNF-α protein(r=-0.541,P<0.01). Conclusion: The mechanism of Ailuo Kechuanning regulating COPD airway mucus hypersecretion may increase AQP5 gene and inhibit p38 MAPK,TNF-α genes expression.