Tuina ameliorates neurobehavioral deficits and neuroinflammation in a mouse model of cerebral palsy: Associations with metabolic reprogramming.

BACKGROUND

Cerebral palsy (CP) is characterized by persistent motor impairment and may be accompanied by sustained neuroinflammation. Metabolic reprogramming is increasingly recognized as an important regulator of microglial inflammatory responses; however, its involvement in the effects of Tuina remains unclear.

OBJECTIVE

To evaluate the effects of Tuina in a neonatal hypoxic-ischemic mouse model of CP and determine whether its effects are accompanied by changes in cortical energy metabolism and microglial inflammatory profiles.

METHODS

Mice were assigned to Sham, CP, and CP+Tuina groups. Tuina was administered for 15 min/day, 6 days/week, for 7 weeks. Growth, neurobehavioral performance, and brain histopathology were assessed. Cortical protein lactylation profiles, glycolytic activity, mitochondrial respiration, metabolic enzyme activities, and ATP content were evaluated. Microglia-associated metabolic alterations were examined by double immunofluorescence staining of Iba1 with GLUT1 or COX IV. Activation-associated markers and inflammatory cytokines were also measured. In vitro, BV2 microglial cells were treated with oxamate or rotenone to modulate glycolysis and mitochondrial respiration, respectively.

RESULTS

Tuina improved body-weight gain, motor coordination, cognitive performance, and histopathological changes in CP mice. Protein lactylation profiling identified alterations in pathways related to glycolysis, the tricarboxylic acid cycle, and oxidative phosphorylation. CP mice exhibited increased cortical glycolytic activity and impaired mitochondrial respiration, which were partially attenuated by Tuina. In Iba1-positive microglia, CP was associated with increased GLUT1 and decreased COX IV expression, whereas Tuina partially reversed these changes. Tuina also reduced CD86 and iNOS expression and the levels of TNF-α and IL-1β, while further increasing CD206, Arg1, IL-10, and IL-4. In BV2 cells, glycolysis inhibition reduced pro-inflammatory-associated responses, whereas mitochondrial inhibition induced glycolytic compensation and enhanced pro-inflammatory-associated markers and cytokines.

CONCLUSION

Tuina was associated with improved neurobehavioral outcomes, partial restoration of cortical metabolic homeostasis, and modulation of microglial inflammatory profiles in CP mice. The findings support a close relationship between metabolic reprogramming and microglia-mediated inflammation, although the causal role of these metabolic changes in the in vivo effects of Tuina requires further investigation.