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Published on: August 15, 2025
Kai-Xin-San improves aging and associated neuroinflammation through mitochondrial autophagy
Yu Gao1, Jinxue Zhang1, Xiaoqian Zhang1
1School of Pharmacy, Center for Xin'an Medicine and Modernization of Traditional Chinese Medicine of IHM, Anhui University of Chinese Medicine, Hefei, 230012, Anhui, China; Anhui Province Key Laboratory of Pharmaceutical Preparation Technology and Application, Institute of Pharmacokinetics, Hefei, 230012, Anhui, China; Engineering Technology Research Center of Modernized Pharmaceutics, Anhui Education Department (AUCM), Hefei, 230012, Anhui, China; Anhui Academy of Chinese Medicine, Anhui Genuine Chinese Medicinal Materials Quality Improvement Collaborative Innovation Center, Hefei, 230012, Anhui, China.
Ethnopharmacological Relevance:
Kai-Xin-San (KXS), a historically used traditional Chinese prescription, calms the mind, invigorates Qi, removes dampness and turbidity. Certain components of KXS have demonstrated anti-aging and anti-neuroinflammatory properties, but the underlying mechanisms of KXS as a whole remain unclear.
Aim Of The Study:
To investigate the mechanisms through which KXS alleviates aging and neuroinflammation.
Materials And Methods:
Potential targets and signaling pathways of KXS in promoting mitochondrial autophagy and anti-aging were predicted using network pharmacology and molecular docking. In animal experiments, spatial learning memory and neuroinflammation in aging mice following KXS treatment were evaluated through behavioral tests and measurement of biochemical markers. Additionally, histopathological changes in the brain tissue of aged mice, along with mitochondrial autophagy levels, were examined using Hematoxylin and Eosin staining, Nissl staining, immunofluorescence labelling, and transmission electron microscopy (TEM). In cell experiments, the regulatory effect of KXS on mitochondrial autophagy was assessed using TEM and immunofluorescence techniques. Western blotting analyzed aging, inflammatory, and mitochondrial autophagy-related proteins in both models.
Results:
Network pharmacology analysis identified 142 potential targets associated with aging and autophagy. Molecular docking suggested there was strong binding affinity between main components of KXS and relevant targets. In vivo, KXS significantly improved learning memory, biochemical indicators, and neuronal damage, immunofluorescence confirmed that it reduced the expression of the aging marker p21. In vitro, KXS-medicated serum increased mitochondrial autophagy. Moreover, TEM showed that KXS improved mitochondrial damage in aging mice and BV2 cells. Notably, KXS down-regulated the expression aging-related and inflammation-related proteins, then increased the expression mitochondrial autophagy proteins. Importantly, the therapeutic effects of KXS were diminished upon administration of chloroquine (CQ), a mitochondrial autophagy inhibitor.
Conclusion:
KXS could promote in vivo and in vitro mitochondrial autophagy and significantly ameliorate aging and associated neuroinflammation.
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