Objective: To study the protective effect of gastrodin on primary cortical neurons damaged by oxygen-glucose deprivation and reperfusion (OGD/R)

and the impact on expression of inflammation-related signaling pathway. Method: The primary cortical neurons of rats were randomly divided into control group

model group

and high

middle and low-dose gastrodin (80

60

40 mg·L-1).The OGD/R damage model was built by oxygen-glucose deprivation and reperfusion.The survival rate of nerve cells was determinated by 3-(4

5-dimethyl-2-thiazolyl)-2

5-diphengl-2-H-tetrazolium bromide(MTT) method

and the leakage rate of lactate dehydrogenase (LDH) was measured by colorimetric method

high-throughput RNA-Seq transcriptome sequencing analysis was used to detect the differential expressions of all genes in the treated cells

and the OGD/R damage or protection-related genes or/and metabolic pathways were screened by functional annotation and enrichment analysis. Result: The activity of cortical cells decreased and the release of LDH increased after OGD/R

while gastrodin pretreatment increased cell viability and significantly lowered the LDH leakage rate.The sequence analysis revealed that the molecular mechanism mainly involved the inhibition of NF-kappa B signaling pathway (NF-κB

18 gene)

and tumor necrosis factor signaling pathway (TNF

21 gene).Compared with the model group

gastrodin inhibited expressions of NF-κB

Toll like receptor 2 (TLR2)

tumor necrosis factor receptor 1/2 (TNFR1/2)

interleukin-6 (IL-6)

intercellular cell adhesion molecule-1 (ICAM-1)

interferon-inducible protein-10 (IP-10)

monocyte chemoattractant protein-1 (CCL2/MCP-1)

CXC-chemokine 1/2/3 (CXCL1/2/3)

CXC-chemokine 9(CXCL9/Mig)

and other inflammatory signaling molecules. Conclusion: Gastrodin has a protective effect on the cortical neurons of rats after oxygen-glucose deprivation and reperfusion

and its mechanism is mainly related to the inhibition of TLR-NF-κB-TNF inflammatory cascade-related signaling pathways.