清肺颗粒干预TLR4/NF-κB信号通路调控COPD大鼠气道损伤的机制研究  

Effects of Qingfei Granule protecting airway injury of COPD rats by TLR4/NF-κB signaling pathway

作　　者：折哲[1] 廖健杉 李文杰 刘佳玉 赵志翔[1] 刘丹[1] 唐斌擎[1] 张亚达 熊必丹[1] 石克华[1] ZHE Zhe;LIAO Jianshan;LI Wenjie;LIU Jiayu;ZHAO Zhixiang;LIU Dan;TANG Binqing;ZHANG Yada;XIONG Bidan;SHI Kehua(Shanghai Municipal Hospital of Traditional Chinese Medicine Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Jing’an District North Railway Station Hospital,Shanghai 200070,China)

机构地区：[1]上海中医药大学附属市中医医院,上海200071 [2]上海中医药大学,上海201203 [3]静安区北站医院,上海200070

出　　处：《上海中医药杂志》2022年第3期64-73,共10页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市科委“科技创新行动计划”医学创新研究专项面上项目(21Y11920300);上海中医药大学自然科学类基金项目(18LK059)

摘　　要：目的从气道炎症和气道屏障损伤角度探索清肺颗粒通过干预TLR4/NF-κB信号通路调控慢性阻塞性肺疾病(COPD)大鼠气道损伤的作用机制。方法SD大鼠60只,随机选取10只为正常组,其余50只大鼠均采用香烟烟雾(CS)暴露联合气管注射脂多糖(LPS)的方法建立COPD模型。造模成功后,将上述50只大鼠随机分为模型组,地塞米松组(灌胃地塞米松0.2 mg/kg),清肺低、中、高剂组(分别灌胃清肺颗粒5.4 g/kg、10.8 g/kg、21.4 g/kg),每组10只,共灌胃8周。测定大鼠肺功能变化情况,透射电镜下观察肺脏亚显微结构改变;ELISA法测定血清及支气管肺泡灌洗液(BALF)中细胞因子[包括转化生长因子β1(TGF-β1)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)、白介素-10(IL-10)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)、内皮素-1(ET-1)]含量;qRT-PCR法测定肺组织中微小RNA-146a(miRNA-146a)、Toll样受体4(TLR4)、核转录因子-κB(NF-κB)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)mRNA的表达情况;Western blot法测定肺组织TLR4、NF-κB、Bcl-2、Bax的蛋白表达水平。结果(1)与模型组比较,清肺颗粒各组的肺通气功能有所改善,表现为0.2 s用力呼气容积(FEV0.2)/用力肺活量(FVC)水平均有所上升(P<0.05),而RL和Re水平有不同程度下降(P<0.05),且清肺高剂组对大鼠肺功能FEV0.2和FEV0.2/FVC水平的改善程度与地塞米松组相似(P>0.05)。(2)通过透射电子显微镜下观察,与模型组比较,清肺高剂组的Ⅱ型肺泡上皮细胞胞浆质地均匀,核膜未见明显破坏,可见较多线粒体和板层小体,细胞形态大多正常。(3)与模型组比较,清肺颗粒可降低大鼠血清和BALF液中TNF-α、IL-8水平(P<0.05),且中、高剂量组具有一定的剂量依赖关系。(4)与模型组比较,清肺颗粒可上调miRNA-146a mRNA表达,降低大鼠肺组织TLR4、NF-κB、Bax mRNA和其蛋白表达量,上调Bcl-2 mRNA和其蛋白表�Objective To explore the mechanism of Qingfei Granule regulating airway injury in COPD rats by interfering with TLR4/NF-κB signal pathway from the point of view of airway inflammation and airway barrier injury.Methods In 60 SD rats,10 rats were randomly selected as the normal group,and the other 50 rats were exposed to cigarette smoke combined with endotracheal injection of LPS to establish COPD model.After successful establishment of the model,50 rats were randomly divided into the model group,dexamethasone group(dexamethasone 0.2 mg/kg),Qingfei Granule low,medium and high dose groups(Qingfei Granule 5.4 g/kg,10.8 g/kg,21.4 g/kg,respectively),with 10 rats in each group and were respectively given the corresponding drugs by intragastric administration for 8 weeks.The changes of pulmonary function and ultrastructure of lung were observed under transmission electron microscope.The contents of cytokines(including TGF-β1,TNF-α,IL-1β,IL-6,IL-10,VEGF,CTGF and ET-1)in serum and bronchoalveolar lavage fluid(BALF)were measured by ELISA.The expressions of TLR4,NF-κB,Bcl-2 and Bax mRNA in lung tissue were measured by qRT-PCR.The protein expressions of TLR4,NF-κB,Bcl-2 and Bax in lung tissue were measured by Western blot.Results(1)Compared with the model group,the pulmonary ventilation function of different Qingfei Granule groups was improved,showing that the level of FEV0.2/FVC was increased(P<0.05),while the levels of RL and Re decreased in different degrees.The improvement degree of FEV0.2 and FEV0.2/FVC in high dose group of Qingfei Granule was similar to that in dexamethasone group(P>0.05).(2)Through the observation under transmission electron microscope,compared with the model group,the cytoplasmic texture of typeⅡalveolar epithelial cells in the high dose group of Qingfei Granule was uniform.There was no obvious destruction of nuclear membrane.More mitochondria and lamellar bodies could be seen,and most of the cells were normal.(3)Compared with the model group,Qingfei Granule could reduce the levels of TNF-αa

关 键 词：TLR4/NF-κB信号通路 凋亡与修复 机制研究 模型大鼠 中药研究 

分 类 号：R285.5[医药卫生—中药学]