基于MAPK/ERK/Nrf2信号通路的参七虫草方对肺纤维化大鼠免疫串化的动态实验研究  

Dynamic experimental study on the effect of Shenqi Chongcao Formula on immune tandem reaction in IPF model rats based on the MAPK/ERK/Nrf2 signal pathway

作　　者：王三凤 陈炜[2] 何程 栾军 WANG Sanfeng;CHEN Wei;HE Cheng;LUAN Jun(Anhui University of Chinese Medicine,Hefei 230038,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学研究生院,合肥230038 [2]安徽中医药大学第一附属医院呼吸内科,合肥230031

出　　处：《天津中医药大学学报》2024年第8期713-722,共10页Journal of Tianjin University of Traditional Chinese Medicine

基　　金：国家中医药管理局青年岐黄学者人才支持项目(国中医药人教发〔2020〕7号文);安徽省临床医学研究转化专项项目(202304295107020106);安徽省教育厅高校自然科学研究重点项目(KJ2021A0554);2022年安徽省中医药传承创新项目(2022CCWT02);安徽省中医药高水平传承人才支持项目(皖卫传(2023)451号)。

摘　　要：[目的]基于MAPK/ERK/Nrf2信号通路,观察国医大师韩明向所创参七虫草方对特发性肺纤维化(IPF)模型大鼠免疫串化的影响。[方法]SPF级SD大鼠150只随机分为五组,空白组、模型组、参七虫草组、吡非尼酮组及Nrf2抑制剂组,每组各30只大鼠。除空白组外均构建IPF大鼠模型,于造模后的第7、14、21、28天称取大鼠体质量并处死各组中的6~8只大鼠,HE、Masson染色观察大鼠肺组织病理变化;ELISA法检测大鼠血清核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)的含量;RT-PCR法检测大鼠肺组织丝裂原活化蛋白激酶1(MEK1)、丝裂原活化蛋白激酶2(MEK2)、细胞外调节蛋白激酶1(ERK1)、细胞外调节蛋白激酶2(ERK2)、衔接蛋白复合物-1(AP-1)mRNA的表达水平;流式细胞术检测各组大鼠肺组织M1及M2型巨噬细胞的阳性率后计算M1/M2,评价各组大鼠的免疫反应程度。[结果]同时间点下相较于空白组,其余四组大鼠于造模后第7、14、21、28天的肺组织均有不同程度的炎性细胞浸润和胶原纤维沉积,第14、21、28天的体质量均较轻(P<0.05),第7、14、21、28天的血清Nrf2、HO-1含量及肺组织MEK1、MEK2、ERK1、ERK2、AP-1 mRNA的相对表达量均明显升高(P<0.05);相较于模型组,药物干预的三组大鼠第14、21、28天的体质量较重,第7、14、21、28天的肺组织病变程度较轻、血清Nrf2、HO-1含量及肺组织MEK1、MEK2、ERK1、ERK2、AP-1 mRNA的相对表达量较低(P<0.05)。相较于空白组,模型组大鼠早期肺组织M1/M2明显升高(P<0.05),后期M1/M2明显降低(P<0.05);相较于模型组,药物干预的三组大鼠早期肺组织M1/M2明显降低(P<0.05),后期M1/M2明显升高(P<0.05)。[结论]参七虫草方同吡非尼酮和Nrf2抑制剂,能够降低大鼠肺纤维化程度,其机制可能为通过抑制MAPK/ERK/Nrf2信号通路,降低AP-1、HO-1的表达水平,从而干预IPF大鼠肺部的免疫反应,减轻炎症损伤,减少胶原沉积,延缓肺纤维化进[Objective]To observe the effect of Chinese medical master HAN Ming-xiang’Shenqi Chongcao Formula on the idiopathic pulmonary fibrosis(IPF)rats based on the mitogen activated protein kinase(MAPK)/extracellular regulated protein kinases(ERK)/nuclear factor-erythroid 2-related factor 2(Nrf2)signal pathway.[Methods]150 Specific Pathogen-Free(SPF)-grade Sprague-Dawley(SD)rats were randomly and evenly divided into 5 groups:the blank group,the model group,Shenqi Chongcao group,Pirfenidone group and the inhibitor of nuclear factor erythroid 2-related factor 2(Nrf2)group.IPF rat models were established except the blank group,then 6~8 rats in each group were weighed and euthanized on the 7th,14th,21st,and 28th days of the experiment.Hematoxylin-eosin(HE)and Masson staining were used to observe the pathological changes in the lung tissues;ELISA method was used to detect the contents of nuclear Nrf2 and heme oxygenase-1(HO-1)in the rats,serum;Reverse Transcription-Polymerase Chain Reaction(RT-PCR)was used to detect the expression levels of mitogen-activated protein kinase 1(MEK1),mitogen-activated protein kinase 2(MEK2),extracellular regulated protein kinases 1(ERK1),extracellular regulated protein kinases 2(ERK2)and adaptor protein complex-1(AP-1)mRNA in the rats,lung tissues;Flow cytometry was used to detect the positive rates of M1 and M2 macrophages,and then M1/M2 was calculated to evaluate the immune response level.[Results]Compared with the blank group and at the same time,on the 14th,21st,and 28th days,the other four groups of rats showed lower weight;on the 7th,14th,21st,and 28th days,the four groups of rats showed varying degrees of inflammatory cell infiltration,collagen fiber deposition in the lung tissue,and higher contents of Nrf2 and HO-1 in the serum,higher expression levels of MEK1,MEK2,ERK1,ERK2 and AP-1 mRNA in the lung tissues.Compared with the model group,on the 14th,21st,and 28th days,the other three groups showed higher weight;on the 7th,14th,21st,and 28th days,the three groups showed milder lesion s

关 键 词：参七虫草方 特发性肺纤维化 免疫反应 巨噬细胞 韩明向 

分 类 号：R285.5[医药卫生—中药学]