基于能量代谢探讨红景清肺方对PM_(2.5)诱导小鼠肺损伤的保护作用  

Protective Effect of Hongjing Qingfei Prescription(红景清肺方)on PM_(2.5)-induced Lung Injury in Mice Based on Energy Metabolism

作　　者：苏鑫[1] 胡万利 张胜君 SU Xin;HU Wanli;ZHANG Shengjun(Changchun University of Chinese Medicine,Changchun 130117,Jilin,China)

机构地区：[1]长春中医药大学,吉林长春130117

出　　处：《中华中医药学刊》2024年第1期5-8,I0002,I0003,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：吉林省教育厅“十三五”科学技术研究项目(JJKH20200888KJ)。

摘　　要：目的探究红景清肺方对大气细颗粒物(particulate matter 2.5,PM_(2.5))诱导小鼠肺损伤的保护作用及机制。方法40只ICR雄性小鼠,随机分为空白对照组、PM_(2.5)模型组、中药等效组、中药二倍组。中药各组预给药7 d后,除空白对照组外,各组持续2 d给予0.02 mg·μL^(-1)浓度PM_(2.5)混悬液滴鼻制备小鼠肺损伤模型。末次染毒24 h后取肺组织样本,苏木素-伊红(hematoxylin-eosin,HE)染色观察肺组织形态学变化;酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测三磷酸腺苷(adenosine triphosphate,ATP)和Na^(+)-K^(+)-ATP酶(Na^(+)-K^(+)-ATPase)、Ca^(2+)-Mg^(2+)-ATP酶(Ca^(2+)-Mg^(2+)-ATPase)含量;蛋白免疫印迹法(western blot)检测腺苷酸活化蛋白激酶(AMP-activatedproteinkinase,AMPK)、过氧化物酶体增殖物激活受体1α(peroxisome proliferator-activated receptor-γcoactivator 1α,PGC-1α)及线粒体转录因子A(mitochondrial transcription factor A,TFAM)蛋白表达水平;免疫组织化学法(immunohistochemistry,IHC)检测PGC-1α、TFAM、核呼吸因子1(nuclear respiratory factor-1,Nrf1)蛋白表达。结果与空白对照组比较,PM_(2.5)模型组小鼠肺组织结构病理损伤明显,肺组织中ATP、Na^(+)-K^(+)-ATP酶、Ca^(2+)-Mg^(2+)-ATP酶含量显著降低(P<0.01),AMPK、PGC-1α、TFAM蛋白表达水平及PGC-1α、Nrf1、TFAM蛋白阳性表达率明显降低(P<0.01);与PM_(2.5)模型组比较,中药各组肺组织损伤明显改善,中药二倍组中ATP含量升高(P<0.05),Na^(+)-K^(+)-ATP酶、Ca^(2+)-Mg^(2+)-ATP酶含量显著升高(P<0.01),中药等效组三者含量有上升趋势,但差异无统计学意义(P>0.05),中药各组AMPK、PGC-1α、TFAM蛋白表达水平及PGC-1α、Nrf1、TFAM蛋白阳性表达率显著升高(P<0.01)。结论红景清肺方可提高肺组织内Na^(+)-K^(+)-ATP酶、Ca^(2+)-Mg^(2+)-ATP酶活性,上调AMPK/PGC-1α信号通路蛋白水平,减轻线粒体损伤和功能障碍,促进线粒体生物合成,有效改善肺组织能量代谢,�Objective This paper investigated the protective effect and mechanism of Hongjing Qingfei Prescription(红景清肺方)on PM_(2.5)-induced lung injury in mice.Methods Totally 40 ICR male mice were randomly assigned into a blank control group,a PM_(2.5)model group,a traditional Chinese medicine equivalent group and a traditional Chinese medicine double group.After 7 days of pre-administration of traditional Chinese medicine,except the blank control group,the other group was given PM_(2.5)suspension(0.02 mg·μL^(-1))continuously for 2 days to prepare a mouse lung injury model through nasal drip.24 hours after the last exposure,the lung tissue samples were taken out and Hematoxylin eosin(HE)staining was used to observe the morphological changes of lung tissue;Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of adenosine triphosphate(ATP),Na^(+)-K^(+)-ATPase and Ca~(2+)-Mg~(2+)-ATPase.Western blot was used to detect AMP activated protein kinase(AMPK)and peroxisome proliferator activated receptor 1α(PGC-1α)and the expression leve of mitochondrial transcription factor A(TFAM)protein.Immunohistochemistry(IHC)was used for detecting protein expressions of PGC-1α,TFAM and nuclear respiratory factor-1(Nrf1).Results Compared with those of the blank control group,the pathological injury of lung tissue structure in the PM_(2.5)model group was obvious,the contents of ATP,Na^(+)-K^(+)-ATPase and Ca^(2+)-Mg^(2+)-ATPase in lung tissues were significantly reduced(P<0.01),and the expression levels of AMPK,PGC-1αand TFAM proteins and the positive expression rates of PGC-1α,Nrf1 and TFAM proteins were significantly reduced(P<0.01).Compared with those of the PM_(2.5)model group,the lung tissue injury in traditional Chinese medicine equivalent group and traditional Chinese medicine double group was significantly improved.The content of ATP in lung tissue in traditional Chinese medicine double group increased(P<0.05),and the contents of Na^(+)-K^(+)-ATPase and Ca^(2+)-Mg^(2+)-ATPase increased significantly(P<

关 键 词：PM_(2.5) 红景清肺方 肺损伤 能量代谢 线粒体 AMPK/PGC-1α信号通路 

分 类 号：R289.5[医药卫生—方剂学]