加味芎蝎散对CVA大鼠肺组织MMP-9和TIMP-1mRNA表达的影响

目的:观察加味芎蝎散对咳嗽变异性哮喘模型鼠肺组织基质金属蛋白酶-9(MMP-9),基质金属蛋白酶抑制剂-1(TIMP-1)mRNA表达的影响。方法:将4月龄的SPF级Wistar大鼠随机分为正常组,模型组,顺尔宁组(1.2 mg·kg-1),加味芎蝎散高、中、低剂量组(8.64,4.32,2.16 g·kg-1)。除正常组外,从第1天开始,各组im 4%卵清蛋白(OVA)溶液0.5 m L,同时ip 2%Al(OH)30.2 m L,每日1次。除正常组外,从第14天开始,各组CSWH 1%OVA溶液攻击2 min,隔日1次,共7次制备模型。造模后第14天起连续ig 15 d,测定咳嗽次数后取材;HE染色观察肺组织病理学改变,瑞氏染色观察肺泡灌洗液炎性细胞变化,RT-q PCR检测肺组织MMP-9,TIMP-1 mRNA的表达。结果:与正常组比较,模型组肺部病理组织损伤和炎性细胞浸润较明显,咳嗽次数明显增加,肺组织MMP-9,TIMP-1 mRNA表达明显升高,均具有统计学差异(P0.01);与模型组比较,加味芎蝎散高、中、低剂量均可改善模型鼠肺部病理损伤及炎性细胞浸润,其中高剂量组最为明显(P0.01),可降低模型鼠咳嗽次数(P0.05,P0.01),下调模型鼠肺组织MMP-9,TIMP-1 mRNA表达(P0.05,P0.01)。结论:加味芎蝎散对咳嗽变异性哮喘气道重塑有一定的抑制作用,其机制可能与下调肺组织MMP-9,TIMP-1 mRNA的表达有关。

Effect of Modified Xiongxie San on mRNA Expressions of MMP-9 and TIMP-1 in Lung Tissue of CVA Rat

Objective: To observe the effect of modified Xiongxie San on mRNA expressions of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in lung tissue of cough variant asthma (CVA) model rat. Method: The 4-month old Wistar rats were randomly divided into the control group, the model group, the singulair group (1.2 mg·kg^-1·d^-1), and the high-, medium-, low-dose modified Xiongxie San groups (8.64, 4.32, 2.16 mg·kg^-1). The CVA model was induced in rats except the rats in the control group by intramuscular injecting of 4% OVA solution of 0.5 mL from the first day, intraperitoneal injecting 2% Al(OH)₃ of 0.2 mL once daily, and inhaling 1%OVA solution for 2 minutes from the fourteenth day once every other one day for 7 times. All rats were administrated intragastrically with the corresponding medicines for continuous 15 days from the fourteenth day. The cough frequency was detected. The pulmonary pathology was observed by HE staining, inflammatory cells in bronchoalveolar lavage fluid(BALF) were observed by Wright's staining. The mRNA expressions of MMP-9 and TIMP-1 in rats' lung tissue were observed by RT-qPCR. Result: Compared with the normal group, the lung injury and inflammatory cell infiltration condition were serious, the cough frequency increased, the mRNA expressions of MMP-9 and TIMP-1 in lung tissue increased in the model group (P <0.01). Compared with the model group, the lung injury and inflammatory cell infiltration condition were improved, the cough frequency decreased, and mRNA expressions of MMP-9 and TIMP-1 in lung tissue decreased in all dose of Xiongxie San groups (P <0.01, P <0.05). Specially, the pulmonary pathology improvement had better result in the high-dose Xiongxie San group (P <0.01). Conclusion: Modifed Xiongxie San could inhibit airway remodeling of CVA, and the pharmacological mechanism may be related to reducing the increased mRNA expressions of MMP-9 and TIMP-1 in lung tissue.