Kaixinsan alleviates adriamycin-induced depression-like behaviors in mice by reducing ferroptosis in the prefrontal cortex

doi:10.12122/j.issn.1673-4254.2024.08.02

Abstract

Abstract:

Objective To investigate the effect of Kaixinsan (KXS, a traditional Chinese medicine formula) for alleviating adriamycin-induced depression-like behaviors in mice bearing breast cancer xenografts and explore the pharmacological mechanism. Methods Forty female BALB/c mice were randomized equally into control group, model group, and low- and high-dose KXS treatment groups, and in the latter 3 groups, mouse models bearing orthotopic breast cancer 4T1 cell xenografts were established and treated with adriamycin along with saline or KXS via gavage. Depression-like behaviors of the mice were assessed using open field test and elevated plus-maze test, and the changes in serum levels of depression-related factors were examined. RNA-seq analysis and transmission electron microscopy were used and ferroptosis-related factors were detected to explore the mechanisms of adriamycin-induced depression and the therapeutic mechanism of KXS. The results were verified in SH-SY5Y cells using ferroptosis inhibitor Fer-1 as the positive control. Results KXS significantly alleviated depression-like behaviors and depression-related serological changes induced by adriamycin in the mouse models. RNA-seq results suggested that KXS alleviated chemotherapy-induced depression by regulating oxidative stress, lipid metabolism and iron ion binding in the prefrontal cortex. Pathological analysis and detection of ferroptosis-related factors showed that KXS significantly reduced ferroptosis in the prefrontal cortex of adriamycin-treated mice. In SH-SY5Y cells, both KXS-medicated serum and the ferroptosis inhibitor were capable of attenuating adriamycin-induced cell ferroptosis. Conclusion KXS alleviates adriamycin-induced depression-like behaviors in mice by reducing ferroptosis in the prefrontal cortex of breast cancer-bearing mice.

Key words: Kaixinsan, breast cancer, chemotherapy, depression, ferroptosis

Mingzi OUYANG, Jiaqi CUI, Hui WANG, Zheng LIANG, Dajin PI, Liguo CHEN, Qianjun CHEN, Yingchao WU. Kaixinsan alleviates adriamycin-induced depression-like behaviors in mice by reducing ferroptosis in the prefrontal cortex[J]. Journal of Southern Medical University, 2024, 44(8): 1441-1449.

Figures/Tables 6

Fig.1 KXS alleviates depression-like behaviors in breast cancer-bearing mice receiving adriamycin treatment. A: Path trace and path heat map in open field test. B: Total distance in open field test. *P<0.05, ***P<0.001 vs control group; ###P<0.001 vs model group; &&P<0.01, &&&P<0.001 vs KXS-L group (n=10).

Fig.2 Effect of KXS on depression-related serological changes induced by adriamycin in breast cancer-bearing mice. The diagrams show comparisons of serum CRH (A), ACTH (B), GC (C), CORT (D), 5-HT (E), NPY (F), NE (G) and BDNF (H) contents among the 4 groups. *P<0.05, **P<0.01, ***P<0.001 vs control group; #P<0.05, ##P<0.01, ###P<0.001 vs model group; &P<0.05, &&P<0.01, &&&P<0.001 vs KXS-L group (n=3).

Fig.3 Target analysis of KXS in alleviating adriamycin-induced depression. A: PCA plot of RNA-seq gene expression. B, C: Volcano plot of differentially expressed genes (DEGs) identified by RNA-seq. D: Venn diagram of the DGEs. E: Cluster heatmap of the common DGEs. F: PPI network of the common DGEs. G: Common DGEs interaction network degree value plot. The top 10 common DGEs are shown on the left.

Fig.4 Target enrichment analysis of KXS in alleviating adriamycin-induced depression. A: GO enrichment analysis of the common DGEs. From top to bottom: molecular function (MF), cellular component (CC), and biological process (BP). B: KEGG pathway enrichment analysis of the common DGEs. C: Reactome pathway enrichment analysis of the common DGEs.

Fig.5 KXS inhibits adriamycin-induced ferroptosis in the prefrontal cortex of the tumor-bearing mice. A: Transmission electron microscopy of the prefrontal cortex tissue of the mice. B: ROS content in the prefrontal cortex tissue. C: Iron content in the prefrontal cortex tissue. D: GSH content in the prefrontal cortex tissue. E: MDA content in the prefrontal cortex tissue. *P<0.05, **P<0.01, ***P<0.001 vs control group; #P<0.05, ##P<0.01, ###P<0.001 vs model group; &&P<0.01 vs KXS-L group (n=3).

Fig.6 Effect of KXS-medicated serum on adriamycin-treated SH-SY5Y cells. A:Growth of SH-SY5Y cells treated with medicated sera for 48 h (Blank: Serum derived from the control group; Adriamycin: Serum from model group containing 10 nmol/L Adriamycin; KXS-L: Serum from low-dose KXS treatment group containing 10 nmol/L Adriamycin; KXS-H: Serum from high-dose KXS treatment group containing 10 nmol/L Adriamycin; Fer-1: Serum from model group containing 10 nmol/L adriamycin and 2 μmol/L Fer-1). B: Cell viability measured by MTT colorimetric assay. C: Lipid ROS content in cell cultures. D: Iron content in the cell cultures. E: GSH content in the cell cultures. F: MDA content in the cell cultures. *P<0.05, **P<0.01, ***P<0.001 vs blank group; ##P<0.01, ###P<0.001 vs adriamycin group; &&P<0.01, &&&P<0.001 vs KXS-L group (n=3).

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