The beneficial properties of Sodium Danshensu (SDSS) for controlling cerebral ischemia and reperfusion injury (CIRI) are elucidated here both in vivo and in vitro. SDSS administration significantly improved the viability of P12 cells, reduced lactate dehydrogenase (LDH) leakage, and decreased the apoptosis rate following exposure to an oxygen-glucose deprivation/reoxygenation (OGD) environment. In addition, the results of a Huprot TM human protein microarray and network pharmacology indicated that AKT1 is one of the main targets of SDSS. Moreover, functional experiments showed that SDSS intervention markedly increased the phosphorylation level of AKT1 and its downstream regulator, mTOR. The binding sites of SDSS to AKT1 protein were confirmed by Autodock software and a surface plasmon resonance experiment, the result of which imply that SDSS targets to the PH domain of AKT1 at ASN-53, ARG-86, and LYS-14 residues. Furthermore, knockdown of AKT1 significantly abolished the role of SDSS in protecting cells from apoptosis and necrosis. Finally, we investigated the curative effect of SDSS in a rat model of CIRI. The results suggest that administration of SDSS significantly reduces CIRI-induced necrosis and apoptosis in brain samples by activating AKT1 protein. In conclusion, SDSS exerts its positive role in alleviating CIRI by binding to the PH domain of AKT1 protein, further resulting in AKT1 activation.

Background: Endothelial pyroptosis promotes cerebral ischemic-reperfusion injury (CIRI). Sodium Danshensu (SDSS) has been shown to attenuate CIRI and have anti-inflammatory properties in endothelial cells. Nevertheless, the mechanism of SDSS on endothelial pyroptosis after CIRI remains unclear. Objective: We aimed to investigate the efficacy and mechanism of SDSS for reducing endothelial pyroptosis. Methods: In vitro, the effect of SDSS alleviating CIRI was first confirmed by detecting pyroptosis and NLRP3 inflammasome related indicators in oxygen-glucose deprivation/reoxygen (OGD/R) treated bEnd3 cells. Further, CLIC4 was identified as a potential target of SDSS through protein microarray, molecular docking, and cellular thermal shift assay (CETSA). Following this, the translocation and expression of CLIC4, and chloride outflow were detected. Finally, CLIC4 was further tested, either overexpressed or knocked down, to determine whether it is a target of SDSS to inhibit endothelial pyroptosis. In vivo, neurological deficit scores and infarct volume were served to evaluate the effect of SDSS in middle cerebral artery occlusion/reperfusion (MCAO/R) rats. Further investigation of pyroptosis was conducted using the CLIC4/NLRP3/GSDMD pathway. Results: SDSS administration inhibited NLRP3 inflammasome mediated pyroptosis in vitro and vivo. As demonstrated by protein microarray, molecular docking and CETSA, SDSS bound strongly to CLIC4 and decreased its protein level, and inhibited its translocation from cytoplasm to cell membrane. Further, SDSS effectiveness was weakened by CLIC4 overexpression but not knockdown. Conclusion: The present study indicated that a beneficial effect of SDSS against CIRI was ascribed to block endothelial pyroptosis via binding to CLIC4, and then inhibiting chloride efflux-dependent NLRP3 inflammasome activation.

Chronic low-grade inflammation has been identified as a major contributor in the development of atherosclerosis. Nuclear Factor-κappa B (NF-κB) is a critical transcription factors family of the inflammatory pathway. As a major catalytic subunit of the IKK complex, IκB kinase β (IKKβ) drives canonical activation of NF-κB and is implicated in the link between inflammation and atherosclerosis, making it a promising therapeutic target. Various natural product derivatives, extracts, and synthetic, show anti-atherogenic potential by inhibiting IKKβ-mediated inflammation. This review focuses on the latest knowledge and current research landscape surrounding anti-atherosclerotic drugs that inhibit IKKβ. There will be more opportunities to fully understand the complex functions of IKKβ in atherogenesis and develop new effective therapies in the future.

Background Traditional Chinese herbal prescription BuYangHuanWu decoction (BHD) works well in treating atherosclerosis. However, the mechanism of BHD in atherosclerosis has not been fully elucidated. We aimed to determine whether BHD could alleviate atherosclerosis by altering the microbiome-associated metabolic changes in atherosclerotic mice. Methods An atherosclerotic model was induced in ApoE−/− mice fed a high-fat diet (HFD), and BHD were administered by gavage for 12 weeks at 8.4g/kg/d and 16.8g/kg/d. Atherosclerotic plaque size, composition, serum lipid profile, as well as inflammatory cytokines were assessed. Mechanistically, metabolomic and micro biota profiling were analyzed using liquid chromatography mass spectrometry (LC-MS) and 16S rRNA gene sequencing, respectively. Further, intestinal microbiota and atherosclerosis-related metabolic parameters were correlated using Spearman analysis. Results Atherosclerotic mice treated with BHD showed a reduction in plaque area, aortic lumen occlusion, and lipid accumulation in the aortic root. 9 perturbed serum metabolites were significantly restored along with the relative abundance of micro biota at the family and genus levels, but not at the phylum level. Gut microbiota improvements were strongly negatively correlated with improved metabolite levels. Conclusions BHD treatment effectively slows the progression of atherosclerosis by regulating altered intestinal microbiota and perturbed metabolites. Detailed analysis of these interactions between intestinal microbes and metabolites affected by BHD will help in the development of new anti-atherosclerosis medications.

Endothelial pyroptosis promotes cerebral ischemia/reperfusion injury (CIRI). Sodium Danshensu (SDSS) has been shown to attenuate CIRI and have anti‐inflammatory properties in endothelial cells. However, the mechanism and effect of SDSS on alleviating endothelial pyroptosis after CIRI remains poorly understood. Thus, we aimed to investigate the efficacy and mechanism of SDSS in reducing endothelial pyroptosis. It has been shown that SDSS administration inhibited NLRP3 inflammasome‐mediated pyroptosis. As demonstrated by protein microarrays, molecular docking, CETSA and ITDRFCETSA, SDSS bound strongly to CLIC4. Furthermore, SDSS can decrease its expression and inhibit its translocation. Its effectiveness was lowered by CLIC4 overexpression but not by knockdown. Overall The beneficial effect of SDSS against CIRI in this study can be ascribed to blocking endothelial pyroptosis by binding to CLIC4 and then inhibiting chloride efflux‐dependent NLRP3 inflammasome activation.