2. Academic Validation
  3. Z-Ligustilide's antidepressant effects in adolescent depression involve the PTEN-induced putative kinase 1 (PINK1) - protein kinase A (PKA)- Na+/Ca2+ exchanger (NCLX) pathway, facilitating mitochondrial endosymbiosis and promoting mitophagy in microglia

Z-Ligustilide's antidepressant effects in adolescent depression involve the PTEN-induced putative kinase 1 (PINK1) - protein kinase A (PKA)- Na+/Ca2+ exchanger (NCLX) pathway, facilitating mitochondrial endosymbiosis and promoting mitophagy in microglia

  • Phytomedicine. 2025 Dec:149:157494. doi: 10.1016/j.phymed.2025.157494.
Tao Yi 1,  Feng Qiu 1,  Jiayi Shi 1,  Dahua Wu 2,  Dandan Li 3,  Xi Zhang 3,  Xu He 4,  Dan Huang 1,  Kaimei Tan 1,  Hongyu Zeng 1,  Yuhong Wang 5,  Jinwen Ge 6,  Zhigang Mei 7,  Xiuli Zhang 8
Affiliations

Affiliations

  • 1 Academy of Chinese Medical Sciences (Institute of Innovation and Applied Research), Hunan University of Chinese Medicine, Changsha 410208, China.
  • 2 Department of Neurology, Hunan University of Chinese Medicine Integrated Chinese Medicine Affiliated Hospital, Changsha, 410208, China.
  • 3 Hunan University of Chinese Medicine, Changsha, 410208, China.
  • 4 School of Basic Medicine, Yiyang Medical College, Yiyang, Hunan, 413000, China.
  • 5 Academy of Chinese Medical Sciences (Institute of Innovation and Applied Research), Hunan University of Chinese Medicine, Changsha 410208, China; The Key Laboratory of Hunan Province for Traditional Chinese Medicine on Preven-tion and Treatment of Depressive Diseases.
  • 6 The Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, Hunan University of Chinese Medicine, Changsha, 410208, China. Electronic address: [email protected].
  • 7 Academy of Chinese Medical Sciences (Institute of Innovation and Applied Research), Hunan University of Chinese Medicine, Changsha 410208, China; The Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, Hunan University of Chinese Medicine, Changsha, 410208, China. Electronic address: [email protected].
  • 8 Academy of Chinese Medical Sciences (Institute of Innovation and Applied Research), Hunan University of Chinese Medicine, Changsha 410208, China; The Key Laboratory of Hunan Province for Traditional Chinese Medicine on Preven-tion and Treatment of Depressive Diseases. Electronic address: [email protected].
PMID: 41218379 DOI: 10.1016/j.phymed.2025.157494
Abstract

Background: Adolescent depression is a prevalent and escalating public health concern. Z-Ligustilide (Z-LIG), a bioactive phthalide extracted from Angelica sinensis (Oliv.) Diels-a Chinese herbal medicine commonly used for its antidepressant effects-exhibits significant anti-inflammatory properties, extensive brain distribution, and low cytotoxicity. Nevertheless, its underlying mechanism in adolescent depression remains unclear.

Purpose: This study aimed to investigate how the PINK1-PKA-NCLX pathway contributes to Z-LIG-induced M2 microglial polarization to alleviate adolescent depression.

Methods: In this study, proteomic analysis followed by molecular docking, surface plasmon resonance (SPR), and tryptophan fluorescence quenching assays identified PINK1 as the target of Z-LIG. This finding was validated using a chronic unpredictable mild stress (CUMS)-induced adolescent rat model. Consequently, the efficacy of Z-LIG in depression-like behaviors and M2 microglial polarization in the hippocampus was confirmed. To explore the role of PINK1-PKA-NCLX pathway in Z-LIG-induced M2 microglial polarization, the NCLX inhibitor CGP-37,157 (CGP) and the PKA Inhibitor dihydrochloride (H89) were used after PINK1 overexpression and Z-LIG treatment. The association between PINK1-PKA-NCLX signaling pathway and PINK1-Parkin pathway was investigated in the context of NCLX overexpression, Z-LIG treatment, and their combination with CGP. Finally, AAV-siPINK1 was administered in vivo to fur corroborate the antidepressant effects of Z-LIG through PINK1 targeting.

Results: PINK1-Parkin-mediated Mitophagy and CA2+ signaling pathways were implicated in adolescent depression. Z-LIG binds strongly to PINK1, thereby enhancing its stability. Z-LIG effectively alleviated depression-like behaviors in adolescent rats by promoting M2 microglia in the hippocampus. PINK1 overexpression and Z-LIG treatment shifted lipopolysaccharide (LPS)-induced M1 microglia to M2 via PINK1-PKA-NCLX and PINK1-Parkin pathways, an effect that was reversed by H89 and CGP. Z-LIG exhibited effects similar to those of NCLX, characterized by reduced cytoplasmic Reactive Oxygen Species (ROS) and DNA levels, while concurrently increasing DNA-mitochondria colocalization and mitochondrial ROS (mtROS) levels in LPS-treated BV2 cells. Furthermore, Z-LIG and NCLX enhanced the AMPK and Parkin phosphorylation. CGP negated the impact of Z-LIG on cytoplasmic ROS, DNA levels, DNA-mitochondria colocalization, and Parkin phosphorylation, but not on mtROS. AAV-si-PINK1 administration abolished the antidepressant effects of Z-LIG.

Conclusions: PINK1-PKA-NCLX pathway contributes to the antidepressant effects of Z-LIG in adolescent depression by stabilizing mitochondrial endosymbiosis and promoting the PINK1-Parkin pathway, which together enhances M2 microglia in the hippocampus. This suggests that Z-LIG may serve as a novel treatment for adolescent depression.

Keywords

Adolescent depression; Microglia; Mitochondrial endosymbiosis; NCLX; Z-Ligustilide (Z-LIG).

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