加味六安煎对咳嗽变异性哮喘模型豚鼠白细胞介素-4、白细胞介素-13水平及气道重塑相关基因表达的影响

Effects of modified Liu’an decoction on IL-4,IL-13 level and expression of relevant genes concerned with airway remodeling in guinea pigs with cough variant asthma model

目的观察加味六安煎对咳嗽变异性哮喘(cough variant asthma,CVA)豚鼠炎症因子白细胞介素(interleukin,IL)-4、IL-13水平及气道重塑相关基因转化生长因子-β(transforming growth factor-β,TGF-β)mRNA、基质金属蛋白酶9(matrix metallopeptidase 9,MMP-9)mRNA表达的影响,探讨加味六安煎治疗咳嗽变异性哮喘的作用机制。方法将30只雄性豚鼠随机分为正常组、CVA模型组、孟鲁司特钠组、加味六安煎低、中、高剂量组(0.74 g/mL、1.48 g/mL、2.22 g/mL),每组各5只。除正常组外,其余组腹腔注射卵蛋白、氢氧化铝致敏并吸入卵蛋白激发法复制咳嗽变异性哮喘豚鼠模型,正常对照组、模型组给予生理盐水,孟鲁司特钠组灌服孟鲁司特钠混悬液(1.2 mg/kg),加味六安煎低、中、高剂量组分别给予不同浓度加味六安煎溶液。酶联免疫法检测血清及支气管肺泡灌洗液(brocho-alveolar larage fluid,BALF)中IL-4和IL-13水平;肺组织胶原面积及胶原沉积率,RT-qPCR法检测肺组织及BALF中的TGF-βmRNA及MMP-9 mRNA的表达;Masson染色法观察并分析肺组织胶原沉积状态。结果(1)与正常对照组相比,模型组血清及BALF中的IL-4水平均降低(P<0.05);与模型组相比,加味六安煎各剂量组(简称:各剂量组)血清中IL-4水平升高(P<0.05)。(2)与正常对照组相比,模型组血清及BALF中的IL-13水平均升高(P<0.05);与模型组相比,各剂量组血清及BALF中IL-13水平均降低(P<0.05)。(3)与正常对照组相比,模型组肺组织及BALF中TGF-βmRNA表达升高(P<0.05);各剂量组肺组织及BALF中TGF-βmRNA表达降低(P<0.05)。(4)与正常对照组相比,模型组肺组织及BALF中MMP-9 mRNA表达升高(P<0.05);各剂量组肺组织及BALF中MMP-9 mRNA表达降低(P<0.05)。(5)与正常对照组相比,模型组胶原面积明显增加(P<0.05);与模型组相比,各剂量组胶原面积减小(P<0.05)。(6)与正常对照组相比,模型组胶原沉积率明显升高(P<0.05);与模型组相比,各剂量组胶原沉积率均有不同程度降低(P<0.05)。结论加味六安煎治疗CVA可能通过调节IL-4和IL-13水平,进而调整辅助型T细胞(T helper cell,Th)Th1/Th2的平衡状态,减轻气道炎症程度;通过调节TGF-βmRNA和MMP-9 mRNA的表达,减轻气道胶原沉积状态,进而改善气道重塑。

Objective To observe the effects of modified Liu’an decoction on expression of interleukin(IL)-4,IL-13 and transforming growth factor-β(TGF-β)mRNA,matrix metallopeptidase 9(MMP-9)mRNA concerned with airway remodeling inguinea pigs with cough variant asthma(CVA),and to explore the mechanism of modified Liu’an decoction in the treatment of CVA.Methods A total of 30 Wistar rats wore randomly allocated into normal group,model group,Montolukast sodium(MS)group,and treatment groups including low-dose,mid-dest and high-dose modified Liu’an Decoction groups.The guinea pig model of CVA was cloned by the method of egg protein and aluminum hydroxide sensitization and inhalation of egg protein stimulation.The inflammatory infiltration of lung tissues of guinea pigs in each group was observed.The IL-4,IL-13 in serum and balf and collagen deposition rate in lung tissues of guinea pigs in each group were compared.Results(1)Compared with the normal group,IL-4 levels in serum of the CVA model group decreased(P<0.05).Compared with the model group,the levels of IL-4 in the treatment groups increased(P<0.05).Compared with the normal group,the level of IL-4 in balf of CVA model group decreased(P<0.05).(2)Compared with the normal group,model group increased significantly(P<0.05),compared with model group,each dose group of modified Liu’an decoction decreased(P<0.05);(3)Compared with the normal group,the tissue and balf of the model group increased(P<0.05),each dose group of modified Liu’an decoction decreased(P<0.05);(4)Compared with the normal group,the tissue and balf of the model group increased(P<0.05),each dose group of modified Liu’an decoction decreased(P<0.05);(5)Compared with the normal group,the tissue and balf of the model group increased(P<0.05),treatment groups decreased(P<0.05);(6)Compared with the normal group,the tissue and balf of the model group increased(P<0.05),treatment groups decreased(P<0.05).Conclusion The treatment of CVA by modified Liu’an decoction may reduce airway inflammation by regulating IL-4 and IL-13 level and then adjusting Th1/Th2 balance,and reduce the degree of airway inflammation;modified Liu’an decoction may improve airway remodeling by adjusting TGF-βmRNA and MMP-9 mRNA so that reduce collagen deposition rate and deposition status of the airway.