Targeting host mitochondrial respiration: artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy <italic style="font-style: italic">via</italic> immunometabolic mechanism

WANG Pan; BAO Chenzhen; ZHU Ting; CEN Yanyan; PAN Xichun

doi:10.16016/j.2097-0927.202512047

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Targeting host mitochondrial respiration: artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy via immunometabolic mechanism

Basic Medicine | 更新时间：2026-03-02

- Targeting host mitochondrial respiration: artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy via immunometabolic mechanism
- Journal of Army Medical University Vol. 48, Issue 4, Pages: 420-432(2026)
- 作者机构：
  
  陆军军医大学（第三军医大学）药学与检验医学系药理学教研室，重庆
- 作者简介：
- 基金信息：
  
  the General Program of the National Natural Science Foundation of China(82373919);the General Project of Natural Science Foundation of Chongqing(CSTB2022NSCQ-MSX0175)
- DOI：10.16016/j.2097-0927.202512047
  CLC： R285.6;R331.142;R392.12
- Received：10 December 2025，
  
  Revised：2026-01-16，
  
  Accepted：16 January 2026，
  
  Published：28 February 2026
- 稿件说明：
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王攀，鲍晨震，朱婷，等.靶向宿主线粒体呼吸：青蒿素衍生物通过上调抗菌性自噬增强巨噬细胞清除细菌的免疫代谢机制［J］.陆军军医大学学报，2026，48（04）：420-432.DOI：10.16016/j.2097-0927.202512047.

WANG P，BAO C Z，ZHU T，et al.Targeting host mitochondrial respiration： artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy via immunometabolic mechanism［J］.J Army Med Univ，2026，48（04）：420-432.DOI：10.16016/j.2097-0927.202512047.
王攀，鲍晨震，朱婷，等.靶向宿主线粒体呼吸：青蒿素衍生物通过上调抗菌性自噬增强巨噬细胞清除细菌的免疫代谢机制［J］.陆军军医大学学报，2026，48（04）：420-432.DOI：10.16016/j.2097-0927.202512047. DOI：

WANG P，BAO C Z，ZHU T，et al.Targeting host mitochondrial respiration： artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy via immunometabolic mechanism［J］.J Army Med Univ，2026，48（04）：420-432.DOI：10.16016/j.2097-0927.202512047. DOI：

摘要

目的

基于宿主线粒体呼吸，探究一种新的青蒿素衍生物（disuccinate artemisinin， DA）通过上调巨噬细胞抗菌性自噬从而促进细菌清除的作用和机制。

方法

实验采用大肠杆菌菌株ATCC 35218和FITC-K12荧光细菌感染小鼠巨噬细胞株RAW264.7，采用荧光成像检测不同时间与不同剂量下巨噬细胞吞噬细菌数量，确定药物适宜剂量与最佳时间点；采用平板菌落计数法、Western blot、免疫荧光共定位等方法，确定药物对巨噬细胞抗菌性自噬的影响；采用自噬抑制剂3-甲基腺嘌呤（3-methyladenine，3-MA）和巴夫洛霉素A1（bafilomycin A1），通过平板菌落计数法、免疫荧光共定位等方法，检测自噬抑制对DA作用的影响；采用低糖培养基对细胞进行葡萄糖剥夺，通过荧光细菌计数和免疫荧光共定位观察葡萄糖剥夺对药物作用的影响；采用糖酵解激活剂寡霉素A（oligomycin A）与糖酵解抑制剂2-脱氧葡萄糖（2-deoxy-d-glucose，2-DG），通过荧光细菌计数和免疫荧光共定位观察糖酵解干预对药物作用的影响；采用线粒体呼吸激活剂碳酰氰-4-三氟甲氧基苯腙（carbonyl cyanide 4-trifluoromethoxy phenylhydrazone，FCCP）和线粒体呼吸抑制剂鱼藤酮（rotenone），通过荧光细菌计数、Western blot共定位观察线粒体呼吸干预对药物作用的影响；通过细胞能量代谢检测巨噬细胞耗氧率（oxygen consumption rate， OCR）确定药物对巨噬细胞线粒体呼吸的影响。

结果

细菌吞噬实验显示，DA可以浓度依赖地促进巨噬细胞吞噬和清除大肠杆菌（

0.05），对耐药菌ATCC 35218和敏感菌K12均有效，最佳工作浓度为10 μg/mL，观察吞噬和清除的时间点分别为1 h和4 h（

0.01）。免疫荧光和免疫印迹结果表明，DA通过上调抗菌性自噬发挥作用（

0.01），在自噬激活抑制时仍有效（

0.01），但自噬流抑制导致其失去作用（

0.01）。在葡萄糖剥夺条件下，DA仍能上调巨噬细胞抗菌性自噬（

0.01）；不论是激活还是抑制糖酵解，均导致DA上调巨噬细胞抗菌性自噬作用受限（

0.01）。激活线粒体最大呼吸后，DA抑制巨噬细胞吞噬细菌并且不能上调抗菌性自噬（

0.01）；阻断线粒体呼吸后，DA仍能上调巨噬细胞吞噬细菌能力和抗菌性自噬（

0.01）。细胞能量代谢分析结果表明，DA可恢复细菌感染导致的巨噬细胞线粒体最大呼吸、线粒体保留呼吸和线粒体呼吸相关ATP生成受损，具有上调线粒体呼吸的作用（

0.01）。

结论

DA通过改善细菌感染时的巨噬细胞线粒体呼吸，从而上调抗菌性自噬以促进清除细菌，是一种靶向线粒体呼吸的宿主导向抗感染候选药物。

Abstract

Objective

To investigate the effect and mechanism of a novel artemisinin derivative

disuccinate artemisinin (DA)

in enhancing bacterial clearance by upregulating antibacterial autophagy in macrophages through targeting host mitochondrial respiration.

Methods

The experiment utilized

E. colistrains

ATCC 35218 and FITC-labeled K12 to infect the mouse macrophage cell line RAW264.7. Phagocytosis of bacteria by macrophages under different time points and doses was assessed

via

fluorescence imaging to determine the optimal drug dosage and time point. The impact of the drug on antibacterial autophagy in macrophages was evaluated using colony counting

Western blotting and immunofluorescence co-localization assay. Autophagy inhibitors 3-methyladenine (3-MA) and bafilomycin A1 were employed to examine the effect of autophagy suppression on drug efficacy through colony counting and immunofluorescence co-localization. Glucose deprivation was induced using low-glucose medium

and its influence on drug action was observed

via

fluorescent bacterial counting and immunofluorescence co-localization. Glycolytic intervention was investigated using the glycolytic activator oligomycin A and inhibitor 2-deoxy-D-glucose (2-DG)

with effects monitored through fluorescent bacterial counting and immunofluorescence co-localization. Mitochondrial respiration was modulated with the activator carbonyl cyanide 4-trifluoromethoxy phenylhydrazone (FCCP) and inhibitor rotenone

and its impact on drug action was assessed

via

fluorescent bacterial counting and immunofluorescence co-

localization. The effect of the drug on mitochondrial respiration in macrophages was determined by measuring the oxygen consumption rate (OCR) using cellular energy metabolism assays.

Results

DA concentration-dependently enhanced macrophage phagocytosis and clearance of both drug-resistant ATCC35218 and sensitive K12

E. coli

(

0.05). Immunofluorescence and immunoblotting demonstrated DA upregulated antibacterial autophagy (

0.01)

remaining effective when autophagy initiation was blocked (

0.01) but losing efficacy upon autophagic flux inhibition (

0.01). Under glucose deprivation

DA retained its autophagy-enhancing effect (

0.01). Both activation and inhibition of glycolysis limited DA’s efficacy (

0.01). Maximizing mitochondrial respiration with FCCP abolished DA’s pro-phagocytic and autophagy-enhancing effects (

0.01)

whereas respiratory blockade with rotenone preserved them (

0.01). Metabolic analysis showed DA restored infection-impaired maximal mitochondrial respiration

spare respiratory capacity

and ATP production (

0.01).

Conclusion

DA enhances bacterial clearance by upregulating antibacterial autophagy through restoration of mitochondrial respiration in infected macrophages

representing a host-directed anti-infective candidate targeting mitochondrial metabolism.

关键词

Keywords

references

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⁰

Targeting host mitochondrial respiration: artemisinin derivative potentiates macrophage bacterial clearance by up-regulating antibacterial autophagy via immunometabolic mechanism

DOI：10.16016/j.2097-0927.202512047

摘要

Abstract

关键词

Keywords

references