Fuzheng Jiedu Tongluo Granule Alleviates Cerebral Ischemia/reperfusion Injury-induced Inflammation By Inhibiting Drp1-mediated TXNIP/NLRP3 Pathway.

Inflammatory response is considered a critical component in developing cerebral ischemia/reperfusion injury (CI/RI). Dynamin-related protein 1 (Drp1) performs an essential role in initiating and advancing CI/RI-induced inflammatory reactions. The objective of the present study was to investigate whether Fuzheng Jiedu Tongluo Granule (FZJDTL) attenuated CI/RI by modulating the Drp1-mediated TXNIP/NLRP3 pathway in vivo and in vitro. Molecular docking and molecular dynamics simulations validated the interactions between the core target and the main components of FZJDTL. Oxygen-glucose deprivation/reoxygenation (OGD/R) in PC12 cells and middle cerebral artery occlusion/reperfusion (MCAO/R) in rats were established. Then, the neuroprotective effects of FZJDTL in MCAO/R rats were assessed using brain infarct volume, HE staining, and neurological deficit scores. The levels of inflammatory factors were detected by ELISA. We further examined whether the neuroprotective effect of FZJDTL on MCAO/R in rats and OGD/R in PC12 cells was connected to the suppression of the Drp1-mediated TXNIP/NLRP3 pathway through methods of Western blot and RT-qPCR. Molecular docking and molecular dynamics simulations revealed that the principal components of FZJDTL exhibited favorable binding affinity with Drp1. The results demonstrated that FZJDTL mitigated pathological brain damage, ameliorated neurological impairments, and reduced the volume of cerebral infarction. FZJDTL inhibited the production of ROS and inflammatory factors (IL-1β, IL-18). In vivo and in vitro studies revealed that the neuroprotective effect of FZJDTL was associated with Drp1-mediated modulation of the inflammatory response, as evidenced by the suppression of mRNA and protein expression of p-Drp1, TXNIP, NLRP3, GSDMD, GSDMD-N, Caspase-1, Cleaved caspase-1, and ASC. Besides, FZJDTL significantly upregulated the mRNA and protein expression of OPA1 and TRX, as verified by RT-qPCR and Western blot analysis. Taken together, treatment with FZJDTL attenuated CI/RI by reducing the inflammatory response via the Drp1-mediated TXNIP/NLRP3 pathway.