清络饮对特发性肺纤维化急性加重大鼠肺组织细胞凋亡相关因子及mRNA的影响  

Effect of Qingluo Yin(清络饮)on Apoptosis Related Factors and mRNA in Lung Tissue of Acute Exacerbation of Idiopathic Pulmonary Fibrosis Rats

作　　者：于睿智 王天娇 臧凝子[1] 吕晓东[2] 庞立健[1] YU Ruizhi;WANG Tianjiao;ZANG Ningzi;LYU Xiaodong;PANG Lijian(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学附属医院,辽宁沈阳110032 [2]辽宁中医药大学,辽宁沈阳110847

出　　处：《辽宁中医药大学学报》2024年第1期36-41,F0003,共7页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82004294);辽宁省“百千万人才工程”资助项目([2017]59);辽宁省科技厅创新能力提升联合基金项目(2022-NLTS-13-3)。

摘　　要：目的 特发性肺纤维化急性加重(AE-IPF)的发病机制尚不明确,现有研究表明AE-IPF的发生与细胞凋亡有关。拟通过探索AE-IPF过程中细胞凋亡相关因子及mRNA的动态表达,以解释基于络病理论所拟复方清络饮通过干预肺组织细胞凋亡进而干预AE-IPF的机制。方法 105只Wistar大鼠雄性采用首次经气管注射博来霉素,2次腹腔注射脂多糖方法进行AE-IPF造模,并将大鼠分为阴性对照组(Control组)、缓解期模型组(IPF组)、急性加重期模型组(AE-IPF组)、中药低剂量组(QLY-L组)、中药中剂量组(QLY-M组)、中药高剂量组(QLY-H组)、激素组(PNS组);Control组、IPF组、AE-IPF组予0.9%生理盐水日2次灌胃,PNS组予醋酸泼尼松日1次灌胃,QLY 3组予低、中、高剂量的中药复方清络饮日2次灌胃。运用HE染色、Masson染色、肺功能检测、支气管肺泡灌洗等方法对大鼠肺功能及肺纤维化程度进行评价,并通过免疫组化、Western-Blot、Quantitative RT-PCR等方法对大鼠肺组织α-SMA、Bax、Bcl-2、Fas、Fas L、Caspase-3进行检测,分析细胞凋亡相关因子及mRNA的表达。结果 予中药复方清络饮的3组大鼠(QLY-L组、QLY-M组、QLY-H组)、予醋酸泼尼松的大鼠(PNS组)和模型组的两组大鼠(IPF组、AE-IPF组)相比,Bcl-2的蛋白(P<0.05)和mRNA表达降低(P<0.05),Bax、Fas、Fas-L、Caspase-3的蛋白(P<0.05)和mRNA表达升高(P<0.05),肺泡炎评分、肺纤维化评分、大鼠肺功能、支气管肺泡灌洗液结果与蛋白和mRNA表达结果趋势一致。结论 中药复方清络饮在调控细胞凋亡、缓解AE-IPF中具有一定疗效,其中清络饮中剂量组疗效较好。Objective The pathogenesis of acute exacerbation of idiopathic pulmonary fibrosis(AE-IPF)is still unclear,and existing studies have shown that the occurrence of AE-IPF is related to apoptosis.This study is to explore the dynamic expression of apoptosis related factors and mRNA in the process of AE-IPF,so as to explain the mechanism of compound Qingluo Yin(清络饮,QLY),which is based on the collateral disease theory,intervening in the apoptosis of lung tissue cells and then intervening in AE-IPF.Methods 105 male Wistar rats were injected bleomycin through air tube for the first time and lipopolysaccharide for the second time to make AE-IPF model.The rats were divided into negative control group(control group),remission model group(IPF group),acute exacerbation model group(AE-IPF group),low dose group(QLY-L group),medium dose group(QLY-M group),high dose group(QLY-H group)and hormone group(PNS group).The control group,IPF group,and AE-IPF group were given 0.9%physiological saline twice daily by gavage,the PNS group was given prednisone acetate once daily by gavage,and the QLY three groups were given low,medium,and high doses of traditional Chinese medicine compound QLY twice daily by gavage.HE staining,Masson staining,pulmonary function detection,bronchoalveolar lavage and other methods were used to evaluate the pulmonary function and pulmonary fibrosis degree of rats,and immunohistochemistry,Western Blot,Quantitative RT-PCR and other methods were used to evaluate the lung tissue of ratsα-SMA,Bax,Bcl-2,Fas,Fas L,Caspase-3 were detected to analyze the expression of apoptosis related factors and mRNA.Results Compared with the three groups of rats(QLY-L group,QLY-M group,QLY-H group),the rats(PNS group)treated with prednisone acetate and the two groups of rats(IPF group,AE IPF group)in the model group,the expression of Bcl-2 protein and mRNA decreased(P<0.05),the protein(P<0.05)and mRNA expression of Bax,Fas,Fas L,Caspase-3 increased(P<0.05).The results of alveolar inflammation score,pulmonary fibrosis score,rat pu

关 键 词：清络饮 AE-IPF 细胞凋亡 α-SMA/Bax/Bcl-2/Fas/Fas L/Caspase-3 大鼠 

分 类 号：R285.5[医药卫生—中药学]