More than microglial depletion: PLX5622 activates the hepatic constitutive androstane receptor to alter anesthesia and addiction

Kelei Cao; Wang Cheng; Liyao Qiu; Zexi Wang; Yuqing Zhao; Ying Yuan; Weiying Wu; Jingyuan Xue; Linghui Zeng; Zhi-Ying Wu; Huan Ma; Tingjun Hou; David A Hume; Cunqi Ye; Shumin Duan; Zhihua Gao

doi:10.1016/j.neuron.2025.12.044

More than microglial depletion: PLX5622 activates the hepatic constitutive androstane receptor to alter anesthesia and addiction

Neuron. 2026 Feb 23:S0896-6273(25)01001-3. doi: 10.1016/j.neuron.2025.12.044. Online ahead of print.

Authors

Kelei Cao¹, Wang Cheng², Liyao Qiu², Zexi Wang², Yuqing Zhao², Ying Yuan², Weiying Wu², Jingyuan Xue³, Linghui Zeng⁴, Zhi-Ying Wu⁵, Huan Ma⁶, Tingjun Hou⁷, David A Hume⁸, Cunqi Ye³, Shumin Duan², Zhihua Gao⁹

Affiliations

¹ Department of Neurology of Second Affiliated Hospital and Liangzhu Laboratory, School of Brain Science and Brain Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China; MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou 311121, China; NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang Key Laboratory of Pain Perception and Neuromodulation, Zhejiang University, Hangzhou 310058, China; Department of Anatomy and Physiology, Shanghai Key Laboratory of Emotions and Affective Disorders, Songjiang Research Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
² Department of Neurology of Second Affiliated Hospital and Liangzhu Laboratory, School of Brain Science and Brain Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China; MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou 311121, China; NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang Key Laboratory of Pain Perception and Neuromodulation, Zhejiang University, Hangzhou 310058, China.
³ Life Sciences Institute, Zhejiang University, Hangzhou 310058, China.
⁴ Key Laboratory of Novel Targets and Drug Study for Neural Repair of Zhejiang Province, School of Medicine, Hangzhou City University, Hangzhou 310015, China.
⁵ Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine and Zhejiang Key Laboratory of Rare Diseases for Precision Medicine and Clinical Translation, Hangzhou 310009, China.
⁶ Department of Neurology of Second Affiliated Hospital and Liangzhu Laboratory, School of Brain Science and Brain Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China; MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou 311121, China; NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang Key Laboratory of Pain Perception and Neuromodulation, Zhejiang University, Hangzhou 310058, China; Affiliated Mental Health Center and Hangzhou Seventh People's Hospital, Zhejiang University School of Medicine, Hangzhou, China.
⁷ Innovation Institute for Artificial Intelligence in Medicine of Zhejiang University, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
⁸ Translational Research Institute, Mater Research Institute, University of Queensland, Woolloongabba, Brisbane, QLD 4102, Australia.
⁹ Department of Neurology of Second Affiliated Hospital and Liangzhu Laboratory, School of Brain Science and Brain Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China; MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, Hangzhou 311121, China; NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang Key Laboratory of Pain Perception and Neuromodulation, Zhejiang University, Hangzhou 310058, China. Electronic address: zhihuagao@zju.edu.cn.

PMID: 41734759
DOI: 10.1016/j.neuron.2025.12.044

Abstract

The colony-stimulating factor 1 receptor (CSF1R) inhibitor PLX5622 has been widely used to deplete microglia for functional characterization and therapeutic support. Although diverse outcomes have been described after PLX5622 treatment, whether these phenotypes solely reflect microglial functions remains to be determined. Here, we show that transgenic microglial depletion did not mimic the accelerated anesthetic arousal or the alleviated nicotine addiction withdrawal symptoms observed after PLX5622 treatment in mice. We further identify that PLX5622 potently activates the mouse constitutive androstane receptor (CAR), leading to prominent induction of hepatic enzymes. The induced enzymatic activity enhances the metabolism and clearance of anesthetics and nicotine, thereby contributing to anesthetic insensitivity and addiction relief. Inactivation of CAR abolished these effects of PLX5622, indicating that the impact of PLX5622 treatment cannot be attributed exclusively to microglial depletion. Our findings raise awareness in evaluating consequences of PLX5622 treatment and provide insights into the design of specific CSF1R inhibitors.

Keywords: PLX5622; anesthesia; colony-stimulating factor 1 receptor; constitutive androstane receptor; metabolism; microglia.