芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠肺组织基质金属蛋白酶9和金属蛋白酶抑制剂1表达的影响
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摘要
目的观察芪蛭皱肺颗粒对慢性阻塞性肺疾病(COPD)模型大鼠肺组织基质金属蛋白酶9(MMP-9)及金属蛋白酶抑制剂1(TIMP-1)表达的影响,探讨其干预气道炎症及气道重塑的作用机制。方法 60只实验大鼠采用气管内滴注脂多糖联合烟熏法复制大鼠COPD模型,第29日开始药物干预,随机分为空白组、模型组、阳性对照组和芪蛭皱肺颗粒高、中、低剂量组,每组10只。第58日,HE染色观察大鼠肺组织形态学变化,ELISA检测大鼠血清MMP-9、TIMP-1含量,Q-PCR和Western blot分别检测大鼠肺组织MMP-9、TIMP-1基因和蛋白表达。结果芪蛭皱肺颗粒各剂量组均可改善COPD大鼠肺组织病理损害。ELISA显示:与空白组比较,模型组大鼠血清MMP-9、TIMP-1含量及MMP-9/TIMP-1比值均显著升高(P<0.05);与模型组比较,芪蛭皱肺颗粒高、中剂量组大鼠血清MMP-9、TIMP-1含量及MMP-9/TIMP-1比值均显著降低(P<0.05)。Q-PCR显示:与空白组比较,模型组大鼠肺组织MMP-9、TIMP-1m RNA表达均显著升高(P<0.05);与模型组比较,芪蛭皱肺颗粒高、中剂量组大鼠肺组织MMP-9 mRNA表达显著降低(P<0.05),芪蛭皱肺颗粒各剂量组大鼠肺组织TIMP-1m RNA表达显著降低(P<0.05)。Westernblot显示,与空白组比较,模型组大鼠肺组织MMP-9、TIMP-1蛋白表达显著升高(P<0.05);与模型组比较,芪蛭皱肺颗粒高、中剂量组MMP-9、TIMP-1蛋白表达显著降低(P<0.05)。结论芪蛭皱肺颗粒可调节大鼠血清和肺组织MMP-9、TIMP-1的表达,纠正MMP-9/TIMP-1状态失衡,抑制肺组织炎症反应,从而干预气道重塑的发生,这可能是其防治COPD的机制之一。
Objective To observe the effects of Qizhi Zhoufei Granules on the expressions of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase 1(TIMP-1) in lung tissues of model rats with chronic obstructive pulmonary disease(COPD); To investigate the mechanism of Qizhi Zhoufei Granules intervening in airway inflammation and airway remodeling. Methods Sixty Wistar rats received intratracheal instillation of lipopolysaccharide combined with smoking to replicate rat models of COPD. Drug intervention started on Day 29. Then the rats were randomly divided into the blank group, model group, positive control group and Qizhi Zhoufei Granules high-, medium-and low-dosage groups, with 10 rats in each group. On day 58, Masson staining was used to observe the change of lung histomorphology. ELISA was used to detect the contents of MMP-9 and TIMP-1 in rat serum. The mRNA and protein expression of MMP-9 and TIMP-1 in lung tissue were detected respectively by Q-PCR and Western blot. Results Qizhi Zhoufei Granules groups could improve the pathological damage of lung tissues in COPD rats. ELISA showed that compared with the blank group, the serum levels of MMP-9, TIMP-1, and MMP-9/TIMP-1 in the model group all increased(P<0.05); compared with the model group, the expressions of MMP-9, TIMP-1, and MMP-9/TIMP-1 in Qizhi Zhoufei Granules high-and medium-dosage groups all decreased(P<0.05). Q-PCR showed that compared with the blank group, the mRNA levels of MMP-9 and TIMP-1 in the lung tissues of model group increased(P<0.05); compared with the model group, the mRNA level of MMP-9 significantly decreased in Qizhi Zhoufei Granules high-and medium-dosage groups(P<0.05), and the m RNA level of TIMP-1 significantly decreased in Qizhi Zhoufei Granules high-, medium-, and low-dosage groups(P<0.05). Western blot showed that compared with the blank group, the protein expressions of MMP-9 and TIMP-1 in the model group increased(P<0.05); compared with the model group, the protein expressions of MMP-9 and TIMP-1 were significantly lower in Qizhi Zhoufei Granules high-and medium-dosage groups(P<0.05). Conclusion Qizhi Zhoufei Granules can regulate expressions of MMP-9 and TIMP-1 and correct the imbalance of MMP-9/TIMP-1, interfere the occurrence and development of lung inflammation in COPD, and then interfere the airway remodeling. This may be one of the mechanisms of Qizhi Zhoufei Granules in the prevention and treatment of COPD.
Objective To observe the effects of Qizhi Zhoufei Granules on the expressions of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase 1(TIMP-1) in lung tissues of model rats with chronic obstructive pulmonary disease(COPD); To investigate the mechanism of Qizhi Zhoufei Granules intervening in airway inflammation and airway remodeling. Methods Sixty Wistar rats received intratracheal instillation of lipopolysaccharide combined with smoking to replicate rat models of COPD. Drug intervention started on Day 29. Then the rats were randomly divided into the blank group, model group, positive control group and Qizhi Zhoufei Granules high-, medium-and low-dosage groups, with 10 rats in each group. On day 58, Masson staining was used to observe the change of lung histomorphology. ELISA was used to detect the contents of MMP-9 and TIMP-1 in rat serum. The mRNA and protein expression of MMP-9 and TIMP-1 in lung tissue were detected respectively by Q-PCR and Western blot. Results Qizhi Zhoufei Granules groups could improve the pathological damage of lung tissues in COPD rats. ELISA showed that compared with the blank group, the serum levels of MMP-9, TIMP-1, and MMP-9/TIMP-1 in the model group all increased(P<0.05); compared with the model group, the expressions of MMP-9, TIMP-1, and MMP-9/TIMP-1 in Qizhi Zhoufei Granules high-and medium-dosage groups all decreased(P<0.05). Q-PCR showed that compared with the blank group, the mRNA levels of MMP-9 and TIMP-1 in the lung tissues of model group increased(P<0.05); compared with the model group, the mRNA level of MMP-9 significantly decreased in Qizhi Zhoufei Granules high-and medium-dosage groups(P<0.05), and the m RNA level of TIMP-1 significantly decreased in Qizhi Zhoufei Granules high-, medium-, and low-dosage groups(P<0.05). Western blot showed that compared with the blank group, the protein expressions of MMP-9 and TIMP-1 in the model group increased(P<0.05); compared with the model group, the protein expressions of MMP-9 and TIMP-1 were significantly lower in Qizhi Zhoufei Granules high-and medium-dosage groups(P<0.05). Conclusion Qizhi Zhoufei Granules can regulate expressions of MMP-9 and TIMP-1 and correct the imbalance of MMP-9/TIMP-1, interfere the occurrence and development of lung inflammation in COPD, and then interfere the airway remodeling. This may be one of the mechanisms of Qizhi Zhoufei Granules in the prevention and treatment of COPD.
引文
[1]中华医学会呼吸病学分会慢性阻塞性肺疾病学组.慢性阻塞性肺疾病诊治指南(2013年修订版)[J].中华结核与呼吸杂志,2013,36(4):253-264.
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[3]张毅,李娟,李金田,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠IFN-γ/IL-4失衡及免疫脏器指数的影响[J].中成药,2015,37(1):189-191.
[4]李娟,张毅,李金田,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病大鼠氧自由基代谢及肺组织形态学的影响[J].中国中医药信息杂志,2013,20(6):38-40.
[5]张毅,李娟,魏舒畅,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠细胞因子的影响[J].中国中医药信息杂志,2013,20(12):26-28.
[6]宋一平,崔德健,茅培英,等.慢性阻塞性肺疾病大鼠模型的建立及药物干预的影响[J].军医进修学院学报,2001,22(2):99-102.
[7]JOLLY M K,WARD C,EAPEN M S,et al.Epithelial mesechymal transition,a spectrum of tates:Role in lung development,homeostasis and disease[J].Dev Dyn,2017,doi:10.1002/dvdy.24541.
[8]BARENS P J.Inflammatory mechanisms in patients with chronic obstructive pulmonary disease[J].J Allergy Clin Immunol,2016,138(1):16-27.
[9]张葵,张樱,陈翌江,等.参芪补肺汤对肺气虚证慢性阻塞性肺疾病大鼠气道重构中NF-κB和MMP-9,TIMP-1表达的影响[J].中国中药杂志,2008,33(18):2129-2132.
[10]刘燕,颜培正,张庆祥.温阳化饮方通过调节MMP-9活性干预支气管哮喘寒饮蕴肺证气道重塑的机制[J].中华中医药杂志,2018,33(1):279-283.
[11]张子洲,王佳佳.MMP-9和紧密连接蛋白Occludin在慢性阻塞性肺疾病大鼠肺部的表达[J].中国老年学杂志,2018,38(4):930-932.
[12]郭贵州,宋文龙,甘桃梅,等.基于MMP-9/TIMP-1失衡研究补益宗气方干预慢性阻塞性肺疾病大鼠气道重塑的机制[J].中华中医药杂志,2016,31(11):4678-4680.
[2]王成阳,李泽庚.六味补气胶囊对COPD肺气虚证大鼠JAK/STAT通路、MMPs/TIMP的影响[J].中华中医药杂志,2014,29(5):1384-1390.
[3]张毅,李娟,李金田,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠IFN-γ/IL-4失衡及免疫脏器指数的影响[J].中成药,2015,37(1):189-191.
[4]李娟,张毅,李金田,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病大鼠氧自由基代谢及肺组织形态学的影响[J].中国中医药信息杂志,2013,20(6):38-40.
[5]张毅,李娟,魏舒畅,等.芪蛭皱肺颗粒对慢性阻塞性肺疾病模型大鼠细胞因子的影响[J].中国中医药信息杂志,2013,20(12):26-28.
[6]宋一平,崔德健,茅培英,等.慢性阻塞性肺疾病大鼠模型的建立及药物干预的影响[J].军医进修学院学报,2001,22(2):99-102.
[7]JOLLY M K,WARD C,EAPEN M S,et al.Epithelial mesechymal transition,a spectrum of tates:Role in lung development,homeostasis and disease[J].Dev Dyn,2017,doi:10.1002/dvdy.24541.
[8]BARENS P J.Inflammatory mechanisms in patients with chronic obstructive pulmonary disease[J].J Allergy Clin Immunol,2016,138(1):16-27.
[9]张葵,张樱,陈翌江,等.参芪补肺汤对肺气虚证慢性阻塞性肺疾病大鼠气道重构中NF-κB和MMP-9,TIMP-1表达的影响[J].中国中药杂志,2008,33(18):2129-2132.
[10]刘燕,颜培正,张庆祥.温阳化饮方通过调节MMP-9活性干预支气管哮喘寒饮蕴肺证气道重塑的机制[J].中华中医药杂志,2018,33(1):279-283.
[11]张子洲,王佳佳.MMP-9和紧密连接蛋白Occludin在慢性阻塞性肺疾病大鼠肺部的表达[J].中国老年学杂志,2018,38(4):930-932.
[12]郭贵州,宋文龙,甘桃梅,等.基于MMP-9/TIMP-1失衡研究补益宗气方干预慢性阻塞性肺疾病大鼠气道重塑的机制[J].中华中医药杂志,2016,31(11):4678-4680.