Combined Deletion of Mitofusin 2 in Adipose-Mesenchymal Derived Stem Cells and Melatonin Offers Additional Benefits on Protecting the Brain Against Acute Ischemic Stroke in Rat.

Background and Aims: Ischemic stroke (IS) remains the third leading cause of death, and the treatment of acute ischemic stroke (AIS) is still a formidable challenge to clinicians. This study tested the hypothesis that combined silencing Mnf2 gene in adipose-derived mesenchymal stem cells (ADMSCs^sil-Mnf2) and melatonin (Mel) therapy was superior to monotherapy on attenuating the brain infarct volume (BIV) and improving neurological function in AIS rats. Results: In vitro and in vivo studies were conducted. In vitro results showed that as compared with the controls (i.e., ADMSCs/N2a cells), the cellular/protein levels of oxidative stress/reactive oxygen species (ROS)/mitochondrial and DNA damaged/apoptotic/cell stress signaling (tumor necrosis factor [TNF] receptor associated factor 6/ apoptosis signal regulating kinase/MKK^4/7/JUN/ERK^1/2/c-Jun) biomarkers were significantly increased in these cells treated by H₂O₂ that were significantly reversed by ADMSCs^sil-Mnf2 or Mel and further significantly reversed by combined therapy (all p < 0.0001). Animals were categorized into groups 1 (sham-operated control)/2 (AIS)/3 (AIS + Mel)/4 (AIS + ADMSCs^sil-Mnf2)/5 (AIS + Mel-ADMSCs^sil-Mnf2) and euthanized by day 28 after AIS. By day 28, the BIV and the brain infarct area (BIA) were lowest in group 1/highest in group 2/significantly lower in group 5 than in groups 3 and 4/significantly increased in group 4 than in group 3, whereas the neurological function displayed an opposite manner of BIV (all p < 0.0001). The protein expressions of oxidative stress/mitochondrial damaged/apoptotic/inflammatory/cell stress signaling biomarkers displayed an identical pattern, whereas the protein expressions of mitochondrial biogenesis/antioxidants and cellular level of neuronal cells exhibited an opposite manner of BIV among the groups (all p < 0.0001). Innovation and Conclusion: ADMSCs^sil-Mnf2 and Mel combined therapy offered synergic effects on attenuating the BIV/BIA and preserving neurological function in rodents after AIS mainly through suppressing oxidative stress/ROS/inflammatory signalings and upregulating antioxidants. Combined ADMSCs^sil-Mnf2 and Mel therapy offered additional benefits on protecting the brain against AIS in rodents. Antioxid. Redox Signal. 43, 427-447.