Table 5 Summarizing the data of all preclinical studies in post-stroke specifying the design, objectives, methodology, results, and conclusion

76

Animal study (MCAO rats and OGD/R cells)

Investigate effects of 10-Hz rTMS on microglial activation and immune pathways

rTMS applied to MCAO rats and OGD/R BV2 cells. Analysis of let-7b-5p levels, HMGA2, NF-kB signaling

Increased let-7b-5p modulated HMGA2 and NF-kB signaling, suppressed M1 microglia. Reduced TNF-α, increased IL-10, reduced ischemic volume, and improved neurological outcomes

10-Hz rTMS modulates immune pathways, suppresses neuroinflammation, and improves outcomes in ischemic models

77

Animal study (MCAO rats)

Examine long-term effects of rTMS on microglia polarization and neurogenesis

Long-term rTMS treatment in MCAO rats. Assessed microglia polarization, neuroinflammation markers, and neuronal differentiation

Promoted anti-inflammatory polarization (CD206 +), reduced cleaved caspase-3 and NF-kB/STAT6 activation, enhanced neurogenesis

Long-term rTMS reduces neuroinflammation and promotes neurogenesis post-stroke

78

Animal study (tMCAO rats)

Evaluate effects of high-frequency rTMS (10- and 20-Hz) on microglial phenotypes and neuroinflammation

tMCAO rats treated with 10- and 20-Hz rTMS Measured microglial activation (Iba-1 +, CD68 +, CD206 + markers), JAK2-STAT3 pathway, cytokine levels

20-Hz rTMS suppressed microglial activation, shifted M1 to M2 phenotype, improved neurological function, and reduced white matter injury

20-Hz rTMS effectively mitigates neuroinflammation and improves post-stroke cognitive impairment

79

Animal study (photothrombotic stroke rats)

Investigate effects of continuous theta-burst stimulation (cTBS) on microglial/astrocytic activation and oxidative damage post-stroke

cTBS applied within 3 h post-stroke Assessed microglia/astrocyte polarization, oxidative markers (3-NT, MnSOD), mitochondrial integrity, and infarct volume

Reduced oxidative damage, infarct volume, and neuronal degeneration. Preserved mitochondrial integrity and redox homeostasis

Shifted M1 to M2 and A1 to A2 phenotypes

Early cTBS effectively preserves neurological function and reduces infarct damage by mitigating oxidative stress and promoting reparative pathways

80

Animal study (focal ischemia in mice)

Evaluate the effects of cathodal tDCS on microglia activation, neurogenesis, and recovery in subacute and chronic stroke phases

Cathodal tDCS applied post-focal ischemia Measured neurogenesis, functional recovery, and microglial activation

Promoted functional recovery and neurogenesis. Suppressed microglia activation within a limited therapeutic time window

Cathodal tDCS supports recovery and reduces inflammation in a phase-dependent manner

81

Animal study (sex-dependent response)

Explore sex differences in microglial and astrocytic response to rTMS

Low- and high-frequency rTMS applied to male and female mice. Assessed GFAP + astrocytes and IBA1 + microglia densities near the injury

Female mice showed reduced GFAP + and IBA1 + densities, while male mice exhibited increased densities following rTMS

rTMS treatment elicits sex-dependent differences in glial activation post-injury

82

Animal study (ischemic stroke in rats)

Assess the effects of vagus nerve stimulation (VNS) on microglial phenotypes and inflammatory signaling in ischemic stroke

VNS applied to MCAO rats. Measured TLR4/NF-kB signaling, α7 nicotinic acetylcholine receptor activation, and inflammatory marker expression (iNOS, TNFα, Arg-1, TGFβ)

VNS reduced pro-inflammatory markers (iNOS, TNFα) and increased regulatory markers (Arg-1, TGFβ). Suppressed TLR4/NF-kB signaling via α7 receptor activation

VNS shifts microglia toward a regulatory phenotype, reducing neuroinflammation and promoting recovery post-stroke