宽胸理肺汤合三子养亲汤改善慢阻肺的实验研究  

Kuanxiong Lifei Decoction Combined with Sanzi Yangqin Decoction Alleviate Chronic Obstructive Pulmonary Disease in Rats

作　　者：薛经纬 陶智会[2] 李勇[3] 朱毅 XUE Jingwei;TAO Zhihui;LI Yong;ZHU Yi(Department of Internal Medicine I,Jiading Hospital of Traditional Chinese Medicine,Shanghai 201800,China;Department of Oncology,Shanghai Seventh People's Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200137,China;Department of Gastroenterology,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海市嘉定区中医医院内一科,上海201800 [2]上海中医药大学附属第七人民医院全科,上海200137 [3]上海市中医医院脾胃病科,上海200071

出　　处：《世界中医药》2022年第11期1572-1577,共6页World Chinese Medicine

基　　金：国家自然科学基金项目(81873157);上海市嘉定区中医重点学科项目(2017-ZYZDZK-02);上海市嘉定区卫计委中医药科研课题(2017-KY-ZYY-13)。

摘　　要：目的:观察宽胸理肺汤合三子养亲汤对慢性阻塞性肺疾病(COPD)大鼠线粒体功能及核苷酸结合寡聚化结构域样受体蛋白3炎性小体通路表达的影响,探讨宽胸理肺汤合三子养亲汤对COPD的保护作用及其机制。方法:本实验采用熏香烟加气管注内毒素法制造COPD模型大鼠。将雄性Wistar大鼠随机分成正常对照组(n=7)、COPD组(n=7)、低剂量组(n=7)、中剂量组(n=7)、高剂量组(n=7)、N-乙酰半胱氨酸(N-acetylcysteine,NAC)组(n=7)。采用蛋白质印迹法检测大鼠肺组织核苷酸结合寡聚化结构域样受体蛋白3、凋亡相关斑样蛋白(ASC)、胱天蛋白酶1,酶联免疫吸附试验法检测下游炎症介质(白细胞介素-1β、白细胞介素-18)的含量,MitoSOX Red探针检测大鼠肺组织线粒体活性氧生成情况,ATP检测试剂盒及JC-1检测试剂盒分别检测肺组织ATP含量及线粒体膜电位变化。结果:与对照组比较,COPD模型组大鼠肺组织线粒体ATP合成量显著降低,线粒体活性氧生成显著增加,差异有统计学意义(P<0.01),线粒体膜电位水平有所降低,但差异无统计学意义(P>0.05)。宽胸理肺汤合三子养亲汤呈剂量依赖性升高COPD大鼠肺组织线粒体ATP合成量,降低活性氧生成(P<0.01),但对线粒体膜电位无显著作用。阳性对照药物NAC也可显著升高COPD大鼠肺组织线粒体ATP合成量,降低活性氧生成(P<0.01)。蛋白质印迹结果显示COPD大鼠核苷酸结合寡聚化结构域样受体蛋白3、凋亡相关斑样蛋白、胱天蛋白酶1-p10蛋白表达显著上升,与正常对照组比较,差异有统计学意义(P<0.05)。宽胸理肺汤合三子养亲汤各剂量及NAC干预后,COPD大鼠的核苷酸结合寡聚化结构域样受体蛋白3炎性小体各蛋白表达呈显著下降趋势(P<0.05)。酶联免疫吸附试验结果显示,COPD大鼠白细胞介素-1β、白细胞介素-18表达显著上升,与正常对照组比较,差异有统计学意义(P<0.05)。宽胸理肺汤合三�Objective:To observe the effects of Kuanxiong Lifei Decoction combined with Sanzi Yangqin Decoction on mitochondrial function and expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)in rats with chronic obstructive pulmonary disease(COPD)and to explore the efficacy and mechanism of Kuanxiong Lifei Decoction combined with Sanzi Yangqin Decoction in the treatment of COPD.Methods:COPD model rats were established by smoking cigarette and injection of endotoxin in trachea.Male Wister rats were randomly assigned into a normal control group(n=7),a COPD group(n=7),a low-dose group(n=7),a medium-dose group(n=7),a high-dose group(n=7),and a N-acetylcysteine(NAC)group(n=7).Western blotting was employed to determine the protein levels of NLRP3,apoptosis-associated speck-like protein(ASC),and caspase-1 in the lung tissue of rats.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of the downstream inflammatory cytokines interleukin-1β(IL-1β)and 1β-18(IL-18).MitoSOX red probe was used to detect the production of mitochondrial reactive oxygen species(ROS).ATP and JC-1 assay kits were used to respectively detect the ATP level and mitochondrial membrane potential in the lung tissue.Results:Compared with the control group,the modeling of COPD inhibited the synthesis of mitochondrial ATP(P<0.01),increased the production of ROS(P<0.01),and decreased the mitochondrial membrane potential(P>0.05).Compared with the model group,Kuanxiong Lifei Decoction combined with Sanzi Yangqin Decoction increased the synthesis of mitochondrial ATP and decreased the production of ROS in a dose-dependent manner(P<0.01),but had no significant effect on mitochondrial membrane potential.Similarly,the positive control drug NAC increased the synthesis of mitochondrial ATP and decreased the production of ROS(P<0.01).Western blot showed that the protein levels of NLRP3,ASC,and caspase1-p10 in COPD rats was significantly higher than those in the normal control group.NAC and Kuanxiong Lifei Decoction c

关 键 词：核苷酸结合寡聚化结构域样受体蛋白3炎性小体 宽胸理肺汤 三子养亲汤 慢性阻塞性肺疾病 线粒体活性氧 香烟 内毒素 实验研究 

分 类 号：R289.5[医药卫生—方剂学] R563[医药卫生—中药学]