Jujuboside A improves insomnia by maintaining mitochondrial homeostasis in prefrontal neurons

Jujuboside A (JB-A) is the major component of Semen Ziziphi Spinosae (SZS), a traditional Chinese herbal medicine used to treat sleep with clinical efficacy. This is the first study to investigate the effects of JB-A on mitochondrial structure and function in the prefrontal cortex of the insomnia model mice. Young adult C57BL/6 mice were induced to develop insomnia by P-chlorophenylalanine. After 14 d of JB-A treatment via gavage, anxiety level was assessed using the open field and elevated plus maze tests. Next, the mitochondrial metabolic activity and morphological changes in the prefrontal cortex of each group of mice, as well as their effects on mitochondrial membrane potential, oxidative phosphorylation levels, and cytochrome c (Cyt c) content in neurons were measured. In our mouse model, JB-A ameliorated anxiety-like behaviors; up-regulated the membrane potential (Δψm) and had a therapeutic effect on the metabolic activity and damaged microscopic structure of mitochondria in the prefrontal cortex; effectively improved mitochondrial function by increasing the expression of Cyt c oxidase I and IV proteins, ATPase activity, and ATP content; and reduced the accumulation of Cyt c in the neuronal cytoplasm while inhibiting mitochondrial permeability transition pore (mPTP) opening. JB-A can improve insomnia by restoring mitochondrial intracellular oxidative phosphorylation, regulating mPTP to maintain mitochondrial homeostasis, and alleviating structural damage, providing a scientific basis for finding new targets for insomnia treatment. • Jujuboside A treatment significantly alleviated anxiety levels in insomnia mice. • Jujuboside A treatment after insomnia significantly upregulated the mitochondrial membrane potential (ΔΨm) in the prefrontal cortex, improved mitochondrial energy metabolism, and maintained neuronal excitability. • Jujuboside A treatment after insomnia significantly maintained the expression levels of key enzymes in the mitochondrial respiratory chain of the prefrontal cortex, sustaining mitochondrial homeostasis. • Jujuboside A treatment after insomnia significantly alleviated mitochondrial swelling and maintained the integrity of the ultrastructure.