Objective:

This study examined the effects of metformin, vitamin E, and coenzyme Q₁₀ [CoQ₁₀] administered individually and in combination in a neonatal rat model of Neonatal hypoxic-ischemic encephalopathy (HIE) induced on postnatal day [PND] 7 by unilateral carotid artery ligation followed by hypoxia. Behavioral, biochemical, neurological and histopathological parameters were evaluated through postnatal day 14.

Methods:

We analyzed preclinical behavioral, histopathological, infarct volume, biochemical assays change in from multiple animal models, examining mechanisms of neuroprotection, pharmacokinetic profiles, safety data, and therapeutic window analyses for both individual and combination therapy approaches.

Results:

The hypothermia group showed pronounced neurological impairment, motor and cognitive deficits, increased oxidative stress- ↑LPO, ↓SOD, ↓GSH, elevated blood brain barrier permeability, and larger infarct volumes. All treatment groups demonstrated improvement across these measures, with the hypothermia + metformin + vitamin E + CoQ₁₀ combination producing the most robust neuroprotection, combination treatment shows significant (p < 0.05) data as compared to individual treatment groups. Triple therapy improved motor coordination and memory, reduced oxidative injury, lowered Evans Blue extravasation and cerebral edema, and markedly decreased infarct size. Histopathological findings confirmed reduced neuronal degeneration and better preservation of cortical and hippocampal regions.

Conclusions:

The complementary mechanisms and synergistic interactions of metformin along with the therapeutic hypothermia support clinical translation of combination protocols. Overall, metformin, vitamin E and CoQ₁₀ exhibited synergistic neuroprotective effects when used together, outperforming monotherapies. This combination represents a promising adjunct to therapeutic hypothermia for mitigating early hypoxic-ischemic brain injury in neonates.