Effects of water extract of the spleen-brain-related mineral drug Shehanshi on mouse sleep

Abstract

Objective

To investigate the effects of water extract of the spleen-brain-related mineral drug Shehanshi on mouse sleep.

Methods

Shehanshi water extract was subjected to component analysis via X-ray photoelectron spectroscopy. Then, the effects of low-dose (50 mg kg^-1) and high-dose (100 mg kg^-1) Shehanshi water extract on mouse sleep were evaluated through behavioral tests such as pentobarbital sodium subthreshold and above-threshold sleep experiments and autonomic activity experiments. Furthermore, transcriptome sequencing and nontarget metabolomics analysis were performed on the spleen and brain tissues of the mice.

Results

The Shehanshi water extract contains a total of 30 elements and can reduce sleep latency, increase sleep time, and increase the sleep rate of mice. In the open field experiment, the movement distance of the mice decreased, and the central residence time and rest time increased. Immunoinfiltration analysis and immunohistochemical verification of spleen tissue showed that compared with those in the control group, the immune abundance of neutrophils in the administration groups increased (P < 0.05). Transcriptome data analysis revealed that the Atp1b2 gene was located at the intersection of the spleen and brain and was positively correlated with neutrophil expression but negatively correlated with the expression of the metabolite oleic acid in brain tissue. Immunohistochemical results showed that Atp2a3 protein expression decreased and Plcg1 protein expression increased in the high-dose group, and the difference was statistically significant (P < 0.05). The Atp1b2 protein level in the spleen tissue was positively correlated with that in the brain tissue of the mice (R = 0.829, P = 0.038). Western blotting revealed that Atp1b2 protein levels in the brain and spleen increased significantly in the high-dose group (P < 0.05).

Conclusion

The mechanism by which Shehanshi water extract influences sleep may be associated with the expression of genes related to the spleen-brain axis and calcium signaling pathways in brain tissues.