目的 探讨清热解毒扶正方药毒素清通过 AMPK/PGC-1α 信号通路调节线粒体生物发生改善细菌性肺炎 的机制。方法 将 50 只 4～6 周龄的 C57BL/6J 雄性小鼠随机分为 5 组，即空白组、模型组、毒素清高剂量组 （15.6 g·kg-1 ）、毒素清低剂量组（7.8 g·kg-1 ）、头孢曲松组（0.195 g·kg-1 ），每组 10 只。造模完成后 6～8 h 开始 给药干预：毒素清高剂量组灌胃给予 15.6 g·kg-1的毒素清溶液，毒素清低剂量组灌胃给予 7.8 g·kg-1的毒素清 溶液，之后每天早、晚灌胃两次；头孢曲松组腹腔注射头孢曲松（0.195 g·kg-1 ）；空白组和模型组同期分别灌服 等量生理盐水。给药结束后进行一般状况观察、苏木素-伊红（HE）染色观察毒素清对肺组织病理形态的改变， 比色法检测肺组织线粒体呼吸链复合物 I、IV 活性，试剂盒法检测肺组织活性氧（ROS）含量，实时荧光定量聚 合酶链反应（RT-qPCR）检测肺组织线粒体 DNA（mtDNA）拷贝数及 AMPK、PGC-1α、NRF1、TFAM、NLRP3、 IL-1β、IL-6、TNF-α mRNA 表达，蛋白免疫印迹（Western Blot）检测肺组织 p-AMPK/AMPK 比值及 AMPK、 PGC-1α、NRF1 蛋白的表达。结果 与空白组比较，模型组小鼠体质量下降（P＜0.05），肺组织炎症浸润程度 增加，肺组织炎症因子 NLRP3、TNF-α、IL-1β、IL-6 mRNA 表达水平显著升高（P＜0.05，P＜0.01），mtDNA 拷贝数减少（P＜0.05），线粒体呼吸链复合物 I、IV 活性下降（P＜0.05，P＜0.01），ROS 水平升高（P＜0.01）， AMPK、PGC-1α、NRF1、TFAM mRNA 表达水平显著下降（P＜0.01），p-AMPK/AMPK 比值及 PGC-1α、NRF1 蛋白表达水平显著下降（P＜0.05，P＜0.01）；与模型组比较，毒素清可使小鼠体质量增加（P＜0.05），肺组织 炎症损伤明显减轻，NLRP3、TNF-α、IL-1β、IL-6 mRNA 表达水平显著下降（P＜0.05，P＜0.01），mtDNA 拷 贝数升高（P＜0.01），线粒体呼吸链复合物 I、IV 活性升高（P＜0.05，P＜0.01），ROS 水平下降（P＜0.01）， AMPK、PGC-1α、NRF1、TFAM mRNA 表达水平显著升高（P＜0.05，P＜0.01），p-AMPK/AMPK 比值及 PGC- 1α、NRF1 蛋白表达水平显著升高（P＜0.05，P＜0.01）。结论 清热解毒扶正方药毒素清能够改善细菌性肺炎， 其机制可能与调节 AMPK/PGC-1α 信号通路促进线粒体生物发生有关。

Objective To investigate the mechanism of Dusuqing（a formula with the functions of clearing heat， removing toxicity， and reinforcing healthy qi） in ameliorating bacterial pneumonia through regulating mitochondrial biogenesis via the AMPK/PGC-1α signaling pathway. Methods Fifty male C57BL/6J mice，aged 4-6 weeks，were randomly divided into five groups （n=10 per group）：a blank control group，a model group，a high-dose Dusuqing group （15.6 g·kg-1 ）， a low-dose Dusuqing group （7.8 g·kg-1 ）， and a ceftriaxone group （0.195 g·kg-1 ）. Drug interventions commenced 6-8 hours post-modeling. The high- and low-dose Dusuqing groups received oral gavage of Dusuqing solution at respective doses twice daily （morning and evening）. The ceftriaxone group received intraperitoneal injection of ceftriaxone. The blank control and model groups received equivalent volumes of saline by gavage. General condition was monitored. Hematoxylin-eosin （HE） staining was used to observe pathological changes in lung tissue. The activities of mitochondrial respiratory chain complexes I and IV in lung tissue were measured by colorimetric assay. The content of reactive oxygen species （ROS） in lung tissue was detected using a commercial kit. Real-time quantitative polymerase chain reaction （RT-qPCR） was employed to assess mitochondrial DNA （mtDNA） copy number and the mRNA expression levels of AMPK，PGC-1α，nuclear respiratory factor 1（NRF1），mitochondrial transcription factor A （TFAM）， NOD-like receptor protein 3（NLRP3）， interleukin （IL）-1β， IL-6， and tumor necrosis factor- α （TNF-α） in lung tissue. Western Blot analysis was performed to determine the p-AMPK/AMPK ratio and the protein expression levels of AMPK，PGC-1α，and NRF1 in lung tissue. Results Compared with the blank control group，the model group exhibited a significant decrease in body mass （P＜0.05）， increased inflammatory infiltration in lung tissue，significantly elevated mRNA expression levels of inflammatory factors NLRP3，TNF- α，IL-1β，and IL-6 （P＜0.05， P＜0.01）， decreased mtDNA copy number （P＜0.05）， reduced activities of mitochondrial respiratory chain complexes Ⅰ and Ⅳ（P＜0.05，P＜0.01），increased ROS levels （P＜0.01），significantly decreased mRNA expression levels of AMPK，PGC-1α，NRF1，and TFAM （all P＜0.01），and significantly reduced p-AMPK/AMPK ratio and protein expression levels of PGC-1α and NRF1（P＜0.05， P＜0.01）. Compared with the model group， Dusuqing treatment led to increased body mass in mice （P＜0.05），markedly alleviated inflammatory injury in lung tissue，significantly decreased mRNA expression levels of NLRP3，TNF-α，IL-1β，and IL-6（P＜0.05，P＜0.01）， increased mtDNA copy number （P＜0.01），elevated activities of mitochondrial respiratory chain complexes Ⅰ and Ⅳ （P＜0.05，P＜0.01），reduced ROS levels （P＜0.01），significantly increased mRNA expression levels of AMPK， PGC-1α， NRF1， and TFAM （P＜0.05， P＜0.01）， and significantly elevated the p-AMPK/AMPK ratio and protein expression levels of PGC-1α and NRF1（P＜0.05， P＜0.01）. Conclusion The Heat-Clearing， ToxicityRemoving， and Healthy Qi-Reinforcing Formula Dusuqing can ameliorate bacterial pneumonia， and its mechanism may be related to promoting mitochondrial biogenesis via modulating the AMPK/PGC-1α signaling pathway.

R285.5

河南省自然科学基金资助项目（242300421294）。